Efikasnost organskih kiselina u očuvanju kvaliteta obroka za ishranu goveda

This paper presents the efficiency of organic acids (sorbic acid, benzoic acid and palm and coconut oil acids, ammonium propionate, etc.) in prevention of microorganisms activities and undesirable spoilage process of total mixed ration (TMR) for cattle feeding during the summer months. Meals treated with these acids have a lower total number of bacteria and yeasts (cfu/g) for 25 to 78%, are less heated up, and are eaten better. Usage of these acid (1-2 L/t of food) does not require the use of expensive equipment. Purchase costs are compensated by the nutrients less loss and more milk production. Protection of TMR meals by organic acids is justified in the cattle diet once a day and at a time of high summer temperatures.U radu je prikazana efikasnost organskih kiselina (sorbinska, benzijeva, kiseline palminog i koksovog ulja, amonijum propionat i dr.) u sprečavanju aktivnosti mikroorganizama i nepoželjnih procesa kvarenja potpunih mešanih obroka (TMR) za ishranu goveda. Obroci tretirani ovim kiselinama imaju manji ukupan broj bakterija i kvasaca, slabije se greju i bolje konzumiraju. Korišćenje kiselina (1-2 L/t hrane) ne zahteva upotrebu skupe opreme. Troškovi nabavke se nadoknađuju manjim gubitkom hranljivih materija i većom proizvodnjom mleka. Zaštita TMR obroka organskim kiselinama ima opravdanje u ishrani goveda jednom dnevno, posebno u vreme visokih letnjih temperatura

Pathohistological changes in kidney and LDH activity in broiler treated with different doses of ochratoxin A

The three-week long trial was performed on day-old Hybro broilers divided into four groups. After 14 days long preexperimental period, the experimental groups were offered feed contaminated with 0.5, 1.0 and 1.5 ppm OA during 7 respectively. At the end of the trial blood and kidney samples were taken for investigations. In broilers feed with 1.5 ppm of OA histopathological examination of the kidney tissue revealed changes located in proximal tubules. Some cells were dim and swollen. These changes produced particular or total reduction in tubular lumen of kidney. Acute tubular necrosis existed in some of tubulocites in form of small foci. Fragmentation of necrotic mass and presence of fresh red blood cells were also detected. The LDH activity was significantly greater in broilers of experimental groups compared with control group. All presented data indicated that intensity of pathohistological alterations and LDH activity depends upon dietary OTA level. Positive correlation between pathohistological changes and increased LDH activity caused by OTA was noticed. Thus, LDH activity mea- sure could be used as early diagnostic tool in measuring changes caused by OTA

Optimization of laboratory conditions for biosynthesis of type A trichothecenes

Type A trichothecenes, T-2 toxin and diacetoxyscirpenol - DAS, belong to one of the most toxic groups of fusariotoxins. Although larger quantities of them can be found more often in cooler parts of Europe, regarding their metabolic characteristics and the types of illnesses they provoke, it is obvious that even smaller quantities of these toxins can cause serious health disturbances of humans and animals in climatic conditions of Serbia. Having in mind the importance of these substances, the aim of this study was to carry out the optimization of laboratory conditions under which screening of Fusarium spp. isolates from Serbia, regarding T-2 toxin and DAS production, should be done. Four cultures of Fusarium sporotrichioides originating from different regions throughout the world, were under present investigation: ITM-391 (Italy), KF-38/1 (Poland), M-1-1 (Japan) and R-2301 (Germany). According to the previous literature data, all of these isolates were T-2 toxin producers, and some of them were also DAS producers. The influence of medium composition (different C and N atoms sources microelements etc), as well as aeration (in liquid media), on biosynthesis process of these mycotoxins, in vitro conditions was investigated. In the case of most Fusarium sporotrichioides isolates, highest yields of T-2 toxin and DAS were achieved under the conditions of more intense aeration, and with the use of glucose (5 or 20%) as a C atom source. Fermentation in semi-synthetic liquid medium, using a rotary shaker, was more suitable for screening the toxicity of the fungal isolates in pure culture because of shorter period of incubation, more simpler sample preparation, obtaining less interfering materials in crude toxin extracts, and possibility for more precise definition of factors influencing the yield of trichothecenes.Trihoteceni grupe A, T-2 toksin i diacetoksiscirpenol - DAS, predstavljaju jednu od najtoksičnijih grupa fuzariotoksina. Oni se u većim koncentracijama češće mogu naći u hladnijim regionima Evrope, ali, u skladu sa njihovim karakteristikama i vrstama oboljenja koja izazivaju, jasno je da i njihove manje količine mogu dovesti do ozbiljnih zdravstvenih poremećaja kod ljudi i životinja u klimatskim uslovima Srbije. S obzirom na značaj ovih jedinjenja cilj ovog istraživanja je bio da se izvrši optimizacija laboratorijskih uslova u kojima bi se ispitivala sposobnost za biosintezu T-2 toksina i DAS-a kod Fusarium izolata iz Srbije. Istraživanjem su bile obuhvaćene 4 kulture F. sporotrichioides poreklom iz različitih zemalja sveta: ITM-391 (Italija), KF-38/1 (Poljska), M-1-1 (Japan) i R-2301 (Nemačka), za koje je prethodno opisano u literaturi da su producenti T-2 toksina, a neke i DAS-a. Ispitan je uticaj sastava podloge (različiti izvori atoma ugljenika i azota mikroelementi i sl) kao i aeracije (u slučaju tečnih podloga) na proces biosinteze ovih mikotoksina in vitro uslovima. Kod većine izolata najveći prinosi T-2 toksina i DAS-a su dobijeni u uslovima veće aeracije i pri upotrebi glukoze (5 ili 20%) kao izvora ugljenikovog atoma. Fermentacija u tečnoj podlozi se pokazala kao pogodnija metoda za testiranje toksigenosti gljivičnih izolata od prirodnog sterilnog supstrata, zbog kraćeg perioda kultivacije, dobijanja sirovih ekstrakata toksina sa manje pratećih materija, kao i mogućnosti preciznijeg definisanja faktora koji utiču na prinos trihotecena

Pojava i učestalost patogena semena žita i industrijskih biljaka u Srbiji

A total of 41 species of fungi were isolated from seed samples of barley, maize, soybean, and sunflower collected at different locations in Serbia. The majority of detected species occurred on barley (35 of 41 species or 87.8%) comparing to soybean (17 of 41 species or 41.5%), sunflower (16 of 41 species or 39.0%) and maize (15 of 41 species or 36.9%). Species belonging to genera Alternaria, Chaetomium, Epicoccum, Fusarium, Penicillium and Rhizopus were present on seeds of all four plant species. Alternaria species were dominant on soybean, barley and sunflower seeds (85.7%, 84.7% and 76.9%). F. verticillioides and Penicillium spp. were mainly isolated from maize seeds (100 and 92.3% respectively), while other species were isolated up to 38.5% (Chaetomium spp. and Rhizopus spp.). F. graminearum, F. proliferatum, F. poae and F. sporotrichioides were the most common Fusarium species isolated from barley (51.1-93.3%), while on the soybean seeds F. oxysporum (71.4%), F. semitectum (57.1%) and F. sporotrichioides (57.1%) were prevalent. Frequency of Fusarium species on sunflower seeds varied from 7% (F. equiseti, F. graminearum, F. proliferatum and F. subglutinans) to 15.4% (F. verticillioides). Statistically significant negative correlation (r = –0.678*) was determined for the incidence of F. graminearum and Alternaria spp., as well as, Fusarium spp. and Alternaria spp. (r = –0.614*), on barley seeds. The obtained results revealed that seedborne pathogens were present in most seed samples of important cereals and industrial crops grown under different agroecological conditions in Serbia. Some of the identified fungi are potential producers of mycotoxins, thus their presence is important in terms of reduced food safety for humans and animals. Therefore, an early and accurate diagnosis and pathogen surveillance will provide time for the development and the application of disease strategies.Ukupno je izolovana 41 vrsta gljiva iz uzoraka zrna ječma, kukuruza, soje i suncokreta, koji su prikupljeni u različitim lokalitetima u Srbiji. Najveći broj gljiva je utvrđen na zrnu ječma (35 vrsta ili 87,8%) u odnosu na zrno soje (17 vrsta ili 41,5%), suncokreta (16 vrsta ili 39,0%) i kukuruza (15 vrsta ili 36,9%). Vrste rodova Alternaria, Chaetomium, Epicoccum, Fusarium, Penicillium i Rhizopus su utvrđene na zrnu sve četiri biljne vrste. Vrste roda Alternaria su dominirale na zrnu ječma (99,5%), suncokreta (52,9%) i soje (50,0%). F. verticillioides (do 92,0%) i Penicillium spp. (do 57,0%) su najčešće izolovane vrste sa zrna kukuruza, dok su ostale vrste izolovane do 38,5% (Chaetomium spp. and Rhizopus spp.). F. graminearum, F. proliferatum, F. poae i F. sporotrichioides su najčešće Fusarium vrste izolovane sa zrna ječma (51,1-93,3%), dok su na zrnu soje dominirale F. oxysporum (71,4%), F. semitectum (57,1%) i F. sporotrichioides (57,1%). Učestalost Fusarium vrsta na zrnu suncokreta je varirala od 7% (F. equiseti, F. graminearum, F. proliferatum and F. subglutinans) do 15,4% (F. verticillioides). Na zrnu ječma utvđena je statistički značajna negativna korelacija između intenziteta napada F. graminearum i Alternaria spp. (r = –0,678*), kao i između Fusarium spp. i Alternaria spp. (r = –0,614*). Dobijeni rezultati pokazuju da su patogene gljive učestale i u visokom intenzitetu pojave prisutne na zrnu važnih žita i industrijskog bilja gajenih u različitim agroekološkim uslovima u Srbiji. Neke od identifikovanih gljiva su potencijalni proizvođači mikotoksina i njihovo prisustvo je važno zbog smanjene bezbednosti hrane za ljude i životinje. Stoga, rana i pouzdana identifikacija patogenih gljiva i procena njihovog značaja u proizvodnji kukuruza, strnih žita i industrijskih biljaka, omogućuje da se pravovremeno razvije i primeni strategija za njihovo suzbijanje i ublažavanje šteta

Basic mechanisms of the cellular alterations in T-2 toxin poisoning: Influence on the choice and result of the therapy

T-2 mycotoxin, secondary metabolite of Fusarium fungi, is one of the most potent cytotoxic representatives of trichothecene mycotoxin type A. After ingestion, T-2 toxin affects actively dividing cells and irreversible post-mitotic cells. In our experiments, the best protective effects were produced by dexametasone (PI = 3.37) and different methylprednisolone formulations (PI = 2.43-2.64). Significant protective efficacy was shown by nimesulide (PI = 1.44) and N-acethyilcistein (PI = 1.29), but their values were higher in a combination with methylprednisolone (PI = 2.16-2.34). Radioprotector amifostine (WR-2721) expressed good protective effects (PI = 1.26) or/and different absorbent formulations, such as: activated charcoal (PI = 1.13) and various Min-a-zel® powder compounds, which are a well known zeolite clinoptilolite absorbents. Among the five zeolite regimens investigated, only Min-a-zel Plus® showed a significant protective effect (PI = 1.77). In summary, the steroidal anti-inflammatory drugs could be recommended as a regimen of choice for treatment of acute T-2 toxicosis while nonsteroidal anti-inflammatory compounds, different absorbent formulations and their combinations with antioxidants or radioprotectors could be important for the treatment of subacute and chronic T-2 toxin poisonings.T-2 mikotoksin, sekundarni metabolit gljivica iz roda Fusarium, jedan je od najtoksičnijih predstavnika trihotecenskih mikotoksina tipa A. Njegove osnovne osobine, prvenstveno velika stabilnost u prirodi, jeftina proizvodnja, teška detekcija i još uvek nepostojanje adekvatnog antidota čine ga veoma dobrim potencijalnim bojnim otrovom. Posle unošenja, u organizmu otrovane jedinke T-2 toksin se u ćelijama vezuje za receptore na ribozomima i pokreće seriju kaskadnih reakcija koje za posledicu imaju smanjenje stabilnosti gRNK i povećanu ekspresiju proinflamatornih gena koji su između ostalog odgovorni za nastanak anoreksije, gubitak telesne mase imunosupresiju, autoimunih efekata i oštećenje većine tkiva. Toksično oštećenje ciljnih organa, nastalo pod dejstvom T-2 toksina, posledica je njegovog citotoksičnog efekta na labilne ćelije i proinflamatornog efekta na stabilne ćelije u organizmu životinja i ljudi. S obzirom na napred iznete činjenice, jasno je što je u našim istraživanjima najbolji terapijski efekat, kod akutnog trovanja T-2 toksinom, postignut primenom antiinflamatornih lekova steroidne strukture, prvenstveno deksametazona (ZI = 3,37) i različitih oblika metilprednizolona (ZI = 2,43-2,64). Osim toga antiinflamatorni lekovi nesteroidne strukture ispoljili su značajan terapijski efekat, nimesulid (ZI = 1,44) i N-acetlilcistein (ZI = 1,29), ali se njihovo zaštitno dejstvo potencira u kombinaciji sa metilprednisolonom (ZI = 2,16-2,34). Terapijsku efikasnost ispoljili su radioprotektor amifostin (WR-2721) (ZI = 1,26) i/ili različiti apsorbensi. Od primenjenih apsorbenasa, kao što su aktivni ugalj (ZI = 1,13) i različiti oblici Min-a-zel-a®, najveći protektivni efekat ispoljio je Min-a-zel Plus® oblik klinoptiolinskog zeolita (ZI = 1,77). Na osnovu prikazanih rezultata, a u skladu sa činjenicom da je citotoksično i proinflamatorno dejstvo T-2 toksina u direktnoj srazmeri sa njegovom akutnom toksičnošću, u potpunosti je opravdano korišćenje visokih doza antiinflamatornih lekova steroidne strukture u terapiji akutnog trovanja T-2 toksinom. Sa druge strane, u terapiji subakutnih ili hroničnih trovanja T-2 toksinom, preporučuje se upotreba antiinflamatornih lekova nesteroidne strukture, različitih apsorbenasa, ili njihove kombinovane primene sa antioksidansima ili radioprotektorima

Intenzivna pojava vrsta roda Aspergillus kao odgovor na uslove spoljašnje sredine u Srbiji

The frequency and incidence of A. flavus and A. niger on barley, maize, soybean, sunflower and wheat grain, the abundance of European corn borer (Ostrinia nubilalis) moths and their interaction depending on weather conditions in the 2008-2012 period were studied. Under the agroecological conditions of Serbia, the species A. niger is more frequent than A. flavus, and concerning the crop species, its frequency is highest in kernels of sunflower, than soybean, maize, barley and wheat. A. flavus was extremely dominant on all plant species in 2012 regarding its frequency: 100% on soybean, 95.3% on maize, 65.2% on barley, 57.1% on sunflower and 45.8% on wheat. Furthermore, the incidence of A. flavus was higher in 2012 than in previous years. The uncommonly high frequency and incidence of A. flavus infestation of maize grain in 2012 were caused by extremely stressful agrometeorological conditions, high temperatures and drought over the period from flowering to waxy maturity of maize. The precipitation factor (Pf = precipitation sum / average monthly temperature) showed that 2012 was extremely arid in June (Pf = 0.57), July (Pf = 1.45), August (Pf = 0.15) and September (Pf = 1.42). European corn borer (ECB) was a second factor causing intensive occurrence of A. flavus on maize grain in 2012. The maximum flight of ECB moths was recorded as early as in July (5,149) and, as a result of this, high damage and numerous injuries were detected at harvest. Those injuries were covered by visible olive-green powdery colonies typical of A. flavus. In the chronology of A. flavus occurrence, these are the first data on its very high frequency and incidence under the agroecological conditions of Serbia. As intensive infections with A. flavus were rare in the past 50 years, the level of aflatoxins in maize grain was low.U ovom radu su analizirani učestalost i intenzitet napada A. flavus i A. niger na 180 uzoraka zrna ječma, 1138 kukuruza, 283 soje, 89 suncokreta i 79 pšenice, brojnost leptira kukuruzovog plamenca (Ostrinia nubilalis), kao i njihova interakcija u zavisnosti od meteoroloških uslova u periodu 2008-2012. U agroekološkim uslovima u Srbiji vrsta A . niger je učestalija od vrste A. flavus, a u zavisnosti od biljne vrste pojavljuje se najčešće na zrnu suncokreta, zatim soje, kukuruza, ječma i pšenice. A. flavus je izuzetno bila dominantna na svim biljnim vrstama u 2012, kako po učestalosti, tako i intenzitetu napada. Ova vrsta je u 2012. godini utvrđena kod svih uzoraka soje (100%), nešto manje na zrnu kukuruza (95,3%), zatim na ječmu (65,2%) i suncokretu (57,1%) i najmanje na zrnu pšenice (45,8%). Intenzitet napada A. flavus je, također, bila veća u 2012. u poređenju sa prethodnim godinama. Neuobičajeno visoka učestalost i intenzitet napada A. flavus na zrnu kukuruza u 2012. godini bila je uslovljena izuzetno stresnim agrometeorološkim uslovima, visokim temperaturama i sušom, od cvetanja do voštane zrelosti kukuruza. Kišni faktor (Kf = suma padavina / prosečna mesečna temperatura) ukazuje da je te godine bila izuzetno aridna klima u junu (Kf = 0,57), julu (Kf = 1,45), avgustu (Kf = 0,15) i septembru (Kf = 1,42). Kukuruzov plamenac je drugi činilac koji je uslovio intenzivnu pojavu A. flavus na zrnu kukuruza u 2012. godini. Maksimalni let leptira ovog insekta utvrđen je veoma rano, posebno let druge generacije, već u julu 2012. (5.149 jediniki), a kao posledica toga u vreme berbe su utvrđena i brojna oštećenja na kojima je bio vidljiv razvoj maslinasto-zelenih praškastih kolonija koje su tipične za A. flavus. U hronologji pojave A. flavus ovo su prvi podaci o njenoj jako visokoj učestalosti i intenzitetu napada u agroekološkim uslovima u Srbiji. Intenzivna pojava A. flavus je bila retka u proteklih 50 godina zbog čega je bio i nizak nivo aflatoksina u zrnu kukuruzu

Histochemical evaluation of T-2 toxin-induced cardiotoxicity in rats: Semiquantitative analysis

In this study female Wistar rats were treated with T-2 toxin (1 LD50 0.23 mg/kg sc) and sacrificed on days 1, 3, 5, 7, 14, 21, 28 and 60 after the treatment. Control groups of rats were treated by saline (1 ml/kg 0.9% NaCl). At each time-schedule, control groups of animals were sacrificed, too. Pathohistological alterations of the heart were evaluated in whole visual fields stained by haematoxylin and eosin (HE), periodic acid- -Schiff's (PAS), Masson-Trichrom's (MT) and Giemsa (GIM) methods. The changes observed were scored by using semiquantitative grading scale. The heart alterations detected in T-2 toxin-treated animals ranged from focal parenchymal or hyaline degeneration (HE = 2.5 - 4.0; p lt 0.05 vs. control) to diffuse necrosis of muscle cells (HE = 5.0; p lt 0.05 vs. control and 1st day after T-2 treatment). The myofibrils were slightly PAS-positive during the first week of the study (PAS = 2.0 - 3.2; p lt 0.05 vs. control and 1st day after T-2 treatment), while a diffuse distribution of glycogen granules in endo- and perimisium were observed from day 21 to 60 in the whole heart' tissue (PAS = 4.0; p lt 0.05 vs. control and 1st day after T-2 treatment). Massive hemorrhagic foci associated with diffuse accumulation and degranulation of MCs were the most intensive from day 28 to 60 of the study (MT = 5.0; p lt 0.05 vs. control and 1st day after T-2 treatment). During the whole study period, irregular distribution of glycogen granules, intensity and total number of haemorrhages were in correlation with the degree of heart structural lesions, which showed the highest coefficient of correlation (r = 0.8750; p lt 0.001). Our results indicate that basic histohemical methods can be a useful tool for evaluation of T-2 toxin-induced cardiac damage, which is probably a result of complex inflammatory mechanisms, eventually leading to vascular lesions and myocardial necrosis, as well as for some potential cardioprotectors in the future.U ovom radu su ispitani toksični efekti na srcu Wistar pacova akutno trovanih T-2 toksinom. Životinje, jednokratno tretirane T-2 toksinom u dozi od 0,23 mg/kg sc (1 LD50), žrtvovane su 1, 3, 5, 7, 14, 21, 28. i 60. dana posle aplikacije otrova. Kontrolne grupe životinja tretirane su fiziološkim rastvorom (1 ml/kg 0,9% NaCl) i žrtvovane u istim vremenskim intervalima. Procena patohistoloških promena izvršena je na uzorcima tkiva srca, bojenih standardnim histohemijskih metodama: hematoksilin i eozin (HE), Gimza (GIM), perjodna kiselina Schiff-ov reagens (PAS) i Masson trichrom (MT), primenom semikvantitativne analize. U srcu pacova tretiranih T-2 toksinom uočene su promene od fokalne parenhimatozne i hijaline degeneracije miofibrila (HE = 2,5-4,0; r lt 0,05 u poređenju sa kontrolom) do fokalne ili difuzne nekroze mišićnih ćelija (HE = 5,0; r lt 0,05 u poređenju sa kontrolom i 1. danom posle aplikacije T-2 toksina). Tokom prve nedelje ispitivanja miofibrile su bile blago PAS-pozitivne (PAS = 2,0-3,2; r lt 0,05 u poređenju sa kontrolom i 1. danom posle aplikacije T-2 toksina), dok je difuzna distribucija granula glikogena u endo- i perimizijumu zapažena od 21. do 60. dana (PAS = 4,0; p lt 0,05 u poređenju sa kontrolom i 1. danom posle aplikacije T-2 toksina). Masivna hemoragična polja, okružena mnogobrojnim inflamatornim ćelijama, naročito su izražena u periodu od 28. do 60. dana ispitivanja (MT = 5,0; p lt 0,05 u poređenju sa kontrolom i 1. danom posle aplikacije T-2 toksina). Tokom celog perioda ispitivanja, nepravilna distribucija granula glikogena, intenzitet krvarenja i ukupan broj mastocita su bili u korelaciji sa stepenom oštećenja tkiva srca (r = 0,8750; p lt 0,001). Dobijeni rezultati su potvrdili ranije iznetu tezu da su kardiotoksični efekti T-2 toksina verovatno rezultat kompleksnih inflamatornih mehanizama