Effect of Pelleted and Chopped Wheat Straw on the Footpad Dermatitis in Broilers

An experiment was conducted to determine whether it is possible to replace the chopped wheat straw with newly formulated pelleted straw taking into account the prevalence and severity of footpad dermatitis (FPD) in broiler chickens. The study included 300 one-day-old Ross 308 broilers. One group of broilers was reared on chopped wheat straw (the particle length of straw >= 10 cm). The second group was kept on newly formulated pelleted wheat straw. The duration of the trial was 42 days. Moisture content in litter materials and FPD occurrence and severity were assessed weekly. First signs of FPD were observed in 21-day old chickens reared on chopped wheat straw (FPD score = 0.52 +/- 0.10) and a week later in a group of birds kept on pelleted straw (FPD score = 0.34 +/- 0.07). At the end of the experiment in both groups of broilers percentage of severe FPD was high. Severe lesions were significantly (p LT 0.01) more frequent (80%) in broilers reared on chopped wheat straw than on pelleted straw (56%). Although the high moisture content was measured in both litter substrates and the high frequency of severe FPD was estimated in both groups of broilers, pelleted straw showed lower moisture content, occurrence and severity of FPD (2.56 +/- 0.19) comparing to chopped wheat straw (3.20 +/- 0.13) at the end of the experiment. Further research can be directed to the technological improvement of chopped and pelleted straw production or into the imprOvement of handling with this substrate

Evaluation of the efficacy of different feed additives to adsorbe T-2 toxin in vitro

In the trial, in vitro HPTLC - High Performance Thin Layer Chomatography was used to determine the amount of 'free', i.e. unbound or non-decomposed T-2 toxin. Mean adsorption or degradation levels of T-2 toxin in examined feed aditives, in in vitro conditions, ranged from 26.06 to 34.84% and did not significantly differ among used adsorbents: inorganic (Minazel plus - Mz), organic (Mycosorb - Ms) and mixed (Micofyx - Mf). All these additives showed better adsorption ability in the acidic environment (pH3).In vitro ispitivanjem određivana je količina 'slobodnog' tj. nevezanog ili nerazgrađenog T-2 toksina tehnikom tankoslojne hromatografije (HPTLC - High Performance Thin Layer Chromatography). Prosečne vrednosti adsorpcije ili degradacije T-2 toksina ispitivanim adsorbentima u uslovima in vitro nisu se značajnije razlikovale u zavisnosti da li je bio umešan neorganski (Minazel plus, Mz), organski (Mycosorb, Ms) ili mešoviti adsorbent (Micofyx, Mf) i iznosile su od 26,06 do 34,84%. Svi ispitivani adsorbenti su pokazali veću sposobnost adsorpcije ovog toksina u kiseloj sredini (pH 3)

Examination of the efficacy of various feed additives on the pathomorphological changes in broilers treated with T-2 toxin

A 21-day-long experiment was performed on 160 one-day-old 'Ross' broiler chicks. This research was done with the aim of investigating pathomorphological changes in broilers exposed to a relatively small amount of T-2 toxin (2 ppm) and the possibility of prevention and/or alleviating adverse effects of T-2 toxin using various feed additives. Pathohistological examination showed negative consequences of T-2 toxin in all examined organs as degenerative changes developed in small intestine mucosa, enterocites and hepatocites necroses, as well as lymphocites depletion in bursa of Fabricius. Disparately from inorganic (Minazel-plus, Mz) and organic (Mycosorb, Ms) adsorbents, which did not provoke protective effects, in liver, small intestine and bursa of Fabricius of broilers who were given feed with T-2 toxin and mixed adsorbent (Mycofix-plus, Mf), mostly preserved structure of these organs could be noted.Eksperiment je izveden na 160 jednodnevnih brojlerskih pilića provenijencije 'Ross', u trajanju od 21 dan. Istraživanja su bila usmerena na omogućavanje detaljnijeg uvida u patomorfološke promene kod brojlera izloženih dejstvu relativno niskih doza T-2 toksina (2 ppm), kao i mogućnosti prevencije ili ublažavanja štetnih efekata korišćenjem različitih adsorbenata. Patohistološkom analizom isečaka tankog creva, jetre i Fabricijeve burze, uočeni su efekti T-2 toksina u svim ispitivanim organima, u vidu degenerativnih promena na sluznici tankog creva, nekroze enterocita i hepatocita, kao i deplecije limfocita u Fabricijevoj burzi. Za razliku od neorganskog (Minazel-plus, Mz) i organskog (Mycosorb, Ms) adsorbenta čijom primenom nije došlo do zaštitnog efekta, u tkivnim isečcima jetre, creva i Fabricijeve burze kod brojlera koji su putem hrane dobijali T-2 toksin i mešoviti adsorbent (Mycofix, Mf), zapaža se uglavnom očuvana struktura ispitivanih organa

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

Mycotoxins in poultry production

All poultry is sensitive to mycotoxins. This partly depends on the type, age and production categories of poultry, their living conditions and nutritive status and partly on the type, quantity and duration of mycotoxin ingestion. The presence of mycotoxins results in significant health disorders and a decrease in production performances. This leads to considerable economic loss for the poultry industry - either direct losses, i.e. death of the poultry or the indirect ones, i.e. the decrease in body mass, number and quality of eggs, greater food conversion, and immunosuppression. Immunosuppression results in increased sensitivity to infective agents and a bad vaccinal response. Morevover, mycotoxin residues in poultry meat, eggs and products derived from them pose a threat to human health. In order to prevent and reduce the negative implications of mycotoxins in the poultry production, it is necessary to create both global and national strategies for combatting mycotoxins, advance diagnostic techniques and procedures, intensify the control of food quality, introduce new limits on the maximum amount of mycotoxins allowed in food and poultry feed used for certain species and categories of animals, and synchronise it with the European standards.Sva živina je osetljiva na mikotoksine u zavisnosti od vrste, starosne i proizvodne kategorije, uslova ambijenta i nutritivnog statusa, sa jedne strane, i vrste, količine i dužine unošenja mikotoksina, sa druge strane. Prisustvo mikotoksina rezultira značajnim poremećajem zdravlja i padom proizvodnih performansi, a samim tim i značajnim ekonomskim gubicima u živinarskoj industriji, kako direktnim, koji se očituju uginućem živine, tako i indirektnim, u vidu pada telesne mase, broja i kvaliteta jaja, veće konverzije hrane i imunosupresijom. Imunosupresija rezultira povećanom osetljivošću na infektivne agense i lošim vakcinalnim odgovorom. Opasnost po ljudsko zdravlje predstavljaju rezidue mikotoksina u živinskom mesu, jajima i proizvodima dobijenim od njih. Da bi se predupredile i smanjile negativne implikacije mikotoksina u živinarskoj proizvodnji potrebno je formirati kako globalne tako i nacionalne strategije za borbu protiv mikotoksina, unaprediti dijagnostičke tehnike i procedure, pooštriti kontrolu kvaliteta hrane, uvesti nove limite za maksimalne količine mikotoksina u hrani i hranivima za pojedine životinjske vrste i kategorije i uskladiti ih sa evropskim standardima

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

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