As shown before, the intact protein can be found in Rhodnius hemo

As shown before, the intact protein can be found in Rhodnius hemolymph 30 min after its ingestion by the insects ( Staniscuaski et al., 2010). It is not known yet how and where this absorption occurs. In Bombyx mori, it CP-868596 nmr has been demonstrated that the dietary mulberry urease is absorbed from the insect gut into the hemolymph and the presence of urease binding molecule(s) in the gut brush border membrane that would mediate this process was postulated ( Kurahashi et al., 2005). The identity of such molecule(s) has not been investigated so far. Fig. 6 summarizes the possible effects of chemical

modifications on the JBU biological properties investigated here. JBU fed to the insects can follow two pathways: 1) be absorbed into the hemolymph and/or 2) be transported to the insect posterior midgut, where it is digested, generating toxic peptide(s), one of which is Jaburetox (Staniscuaski and Carlini, 2012). Since JBU-Lys is hydrolyzed similarly to the native protein, we postulate that the modification of lysines probably impairs JBU absorption into the hemolymph and/or its action on target tissues, including the Malpighian tubules. In JBU-Ac, on the other hand, the release of toxic peptide(s) upon hydrolysis by insect’s enzymes is blocked, reducing the toxicity of the protein. Since the intact protein can still be absorbed into the hemolymph, a residual toxicity

is observed. There was no significant difference in the lethality between the two derivatives forms of JBU, corroborating the idea that a combinatory effect of both, peptides and intact protein, is relevant to its entomotoxic property. C. ensiformis ureases are complex proteins with learn more several biological activities. The entomotoxic activity

is of great interest, since the search for natural insecticides, with none or reduced threat to the environment or to human health is an attractive alternative to synthetic chemical insecticides for pest management ( Isman, 2006). Altogether, the data herein contributed to our understanding of structure/function of the urease entomotoxic activity and represent an advance on the possible use of ureases and/or their derived-peptides as biological tools in pest management. All the authors declare that: Etomidate the paper has not been previously published in whole or in part and is not currently being considered for publication elsewhere; all authors have contributed significantly to the execution, analysis and writing of the paper. This project was supported by Conselho Nacional de Desenvolvimento Cientifico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nivel Superior (CAPES) and Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS). “
“On page 505, the paragraph starting as “McLachlan et al. (2005), Kekre et al. (2005) and Siedlakowski et al. (2008) studied the effects of Pancratistatin (PST)…” describes the toxins produced by the plant Pancratium littorale, also known as spider lilly.

In 1993, the US Environmental Protection Agency (U S EPA, 1993)

In 1993, the US Environmental Protection Agency (U.S. EPA, 1993) established a cancer risk for N-nitrosodimethylamine (NDMA) of about two orders of magnitude higher than the risk estimate by Health Canada (0.7 ng/kg bodyweight per day) ( Health Canada,

2002), while the rat carcinogenic potency of N-nitrosodiethylamine (NDEA) is about three times that of NDMA ( Lijinsky, 1992). In humans, the average intake of NDEA from food is approximately 1 μg/day ( Scanlan, 1983). However, NDEA is also found in cigarettes and cigarette smoke at 0.0083–0.0405 μg/cigarette, as well as in rolls, buns, muffins and bagels (0.023 μg/100 g), ham (0.149 μg/100 g) and even oysters (0.109 μg/100 g) Ribociclib mw ( Stuff et al., 2009). Thus, it is difficult to predict the exact exposure rate of individuals.

NDEA TGF-beta family is a carcinogen that can induce tumors in a variety of organs of many animal species (Magee and Barnes, 1956, Peto et al., 1991 and Lijinsky, 1992). It requires metabolic activation through P450-catalyzed β-hydroxylation, generating unstable metabolites that alkylate the DNA and thus cause tumor formation (Ribeiro Pinto, 2000). Due to their high expression levels of cytochrome P450 (CYP), hepatocytes represent the most suitable model to investigate CYP induction in relation to drug metabolism. In fact it has been shown that phenobarbital (PB) induces markedly increased expression of several phase I and II enzymes, including several forms of cytochrome P450 (Gonzales, 1989 and Waxman and Azaroff, 1992). In the present work we focused on CYP2B1 and CYP2B2, which are two of the major P450 cytochromes that are induced by PB in rat livers. We correlated N-nitrosodiethylamine genotoxicity (micronuclei and mitotic index) and cytotoxicity (survival,

apoptosis, and necroses rates) in primary cultures of female rat hepatocytes in the presence of PB. Female albino Fischer 344 rats (F-344/DuCrl) of 6–8 weeks of age were used Phosphoribosylglycinamide formyltransferase for the experiments (Charles River Laboratories, Germany). The light/dark regime was 12/12 h, and standard pelleted rat feed and drinking water was supplied ad libitum. Three female Fischer 344 rat livers were used per NDEA concentration. Nitrosamines are able to induce liver tumors not only in male, but also in female rats (Lijinsky, 1992). The study was conducted in compliance with the National Research Council of Austria’s “Guide for the Care and Use of Laboratory Animals”. Hepatocytes were prepared according to the two-step collagenase perfusion method (Eckl and Riegler, 1997). Rats were anesthetized with 200 mg/kg sodium pentobarbital. Following hepatic portal vein cannulation, the livers were perfused for 15 min at a flow rate of 15 mL/min with solution A (142 mM NaCl, 6.7 mM KCl, 10 mM HEPES, pH 7.4). Subsequently, the livers were perfused for 20 min (flow rate 10 mL/min) using 200 mL solution C (buffer A and 5.7 mM CaCl2 at a ratio of 9:1) containing 0.

Therefore, in the present study, we also used acetone as the extr

Therefore, in the present study, we also used acetone as the extraction solvent. However, this procedure might be given to leading an overestimation of the risk because it was expected that acetone had greater extraction capacity compared to the authentic simulant which represents the migration of styrene oligomers from polystyrene into food in general use. In this context, regardless of what the genotoxicity tests result in positive or negative, we can obtain the highest doses, at which the genotoxic responses become equivalent to the spontaneous levels. Then, to avoid overemphasis

of the risk, we can compare these doses and the concentration of the styrene oligomers extracted by simulant and this comparison considering a margin of extraction amount would give us valuable insight to assess the risk of the styrene oligomers extracted from polystyrene. Compared with 50% ethanol, acetone clearly had a greater capacity to extract SDs and STs. The concentrations BMS 354825 of SDs and STs in the test solution were 540 ppm and 13,431 ppm, respectively (total, 13,971 ppm), whereas the maximum total concentration of SDs and STs that can be extracted from polystyrene with 50% ethanol solution is 70 ppb [17]. Therefore, the total concentration of SDs and STs extracted with acetone in the test solution was approximately 200,000 times higher than that extracted Selleck Olaparib with 50% ethanol solution. This result shows that the

capacities of stiripentol solvents to extract styrene oligomers from polystyrene are influenced by the polarity of each solvent. Water, which possesses the highest polarity among solvents listed in Table 5, showed the lowest capacity to extract styrene oligomers. On the other hand, regarding the pattern of compounds extracted from the polystyrene, the ratio of SDs to STs became greater when the polarity of the solvent became higher [19]. In addition, the ratio of SDs to

STs in the test solution used in the present study was not so different from that in the GPPS pellets themselves, showing that acetone extraction allowed cells to be exposed to test samples containing a sufficiently high concentration of SDs and STs in a similar ratio to that found in the original GPPS pellets. Both the Ames test and the in vitro chromosomal aberration test, which are the tests required by the FDA and EFSA for the safety evaluation of food packaging, were negative even when high concentrations of oligomers were used compared to the case of fatty-food simulant, suggesting that the risk of genotoxicity of styrene oligomers migrated from polystyrene food packaging into food is very low. Our results also provide useful data on the clastogenic and polyploidy-inducing potential of styrene oligomers. Because of the low solubility of styrene oligomers in aqueous condition, it was expected that the mixture of styrene oligomers would precipitate out of the culture medium during the in vitro chromosomal aberration test, which indeed occurred at doses of 1250 μg/mL or greater.

Moreover, a transcriptomic analysis of B

granulifera was

Moreover, a transcriptomic analysis of B.

granulifera was included to reveal new peptide sequence present in this sea anemone species. This is the first peptidomic and transcriptomic study of the neurotoxic fractions of these sea anemones, and the first report that compares the overall peptide composition of sea anemones species belonging to two distinct families (Stichodactylidae vs. Actiniidae). We found that the neurotoxic fraction of B. granulifera has richer peptide diversity in relation to S. helianthus, as judging by the more complex reversed-phase profile and the resulting higher number of separated peptide components (156 vs. 113 peptides) and toxic fractions (17 vs. 6). However a similar study of B. cangicum yielded a considerable smaller number of peptide components (81) than B. granulifera, despite both sea anemone species belong to the same genus and their chromatographic profiles share a similar complexity and several similarities, therefore Dasatinib such difference does not seem to arise from the use of different Roxadustat manufacturer mucus extraction methods (immersion in distilled

water vs. electrical stimulation). Our study expanded to 156 the estimated maximal number of peptides in the neurotoxic fraction of sea anemones. We emphasize the term “maximal number” as we showed that venom peptide diversity varies among sea anemone species. Moreover, likewise the previous study [85] we found some apparent venom composition overlaps. Structural studies will confirm whether a single neurotoxic peptide is present in two or more sea anemone species. Peptide toxins previously isolated and characterized from S. helianthus and B. granulifera were identified in the present study, with the exception of ShK [14] and ShPI-1 [22]. These toxins seem to be poorly represented in the S. helianthus exudate so it was not possible to detect them by mass spectrometry. ShK occurs in very low amounts either in freeze-dried mucus or in whole Protein kinase N1 body extract [14], so its purification included a precipitation step by heating the sample at low pH, prior to the chromatographic protocol. Likewise, the isolation of

ShPI-1 comprised a precipitation step (trichloroacetic acid treatment) before the chromatographic separation which included affinity chromatography [22], utilized in many instances as a powerful purification method when the protein of interest is a minor component of a complex mixture [13]. Our study confirmed the presence of a very distinguishable feature among sea anemone species of the genus Bunodosoma, a group of abundant and hydrophobic 4–5 kDa peptides that elute in the last reversed-phase fractions ( Fig. 2 and Fig. 3), so far comprising type 1 sodium channels toxins and APETx-like peptides. The sodium channel toxins are BcIII from B. caissarum [55], Bcg 28.19 and Bcg 30.24 from B. cangicum, BgII and BgIII from B. granulifera. The APETx-like peptides are BcIV from B. caissarum [64], Bcg 31.16, Bcg 28.78, Bcg 25.

Finally, patients were enrolled with a clinical suspicion of seve

Finally, patients were enrolled with a clinical suspicion of severe infection and we did not screen all medical admissions for sepsis criteria. We therefore may have missed potentially eligible subjects. Subsequent studies should take this into consideration. This study confirms that severe sepsis in a sub-Saharan Africa setting has high mortality amongst both HIV-infected and uninfected adults. Establishment on ART confers significant survival benefit in the HIV positive subset, and the high mortality among patients on ART for less than three months underscores the importance

of vigilant clinical follow-up among this group. Patients at risk of death can be identified using simple, objectively measurable ABT-199 datasheet criteria, which following validation amongst other buy VX-809 populations can be used to standardise multi-site interventional trials of sepsis bundles in resource-poor settings. These will

enable the formulation of appropriate evidenced-based local guidelines and clinical trials for the management of critically ill adults in sub-Saharan Africa. We wish to thank the staff on the medical admissions ward at Queen Elizabeth Central Hospital, and the laboratory staff and data entry team at the Malawi-Liverpool-Wellcome Clinical Research Programme. Finally, we thank patients and guardians for their participation in the study. “
“Dengue fever (DF) is transmitted by mosquito bites that introduce dengue virus (DENV) serotypes 1–4. DF is endemic in tropical and subtropical regions, with at least 50 million new cases arising each year.1 Dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS), the severe forms of DF, affect people in nearly two-thirds of the countries worldwide, particularly

those in Southeast Asia.2 DENV serotype 2 (DENV-2) is a risk factor associated with DSS, whereas DENV-1, DENV-3, and DENV-4 are not.3 Phosphatidylinositol diacylglycerol-lyase Patients who develop DHF initially present symptoms similar to DF patients, but they abruptly develop plasma leakage, which manifests as haemoconcentration, ascites, and pleural effusion, and may result in irreversible DSS during the defeverence stage.4 The pathogenesis of DHF is not fully understood, but an alteration of T helper (Th)1/Th2 immune reaction is thought to be involved.5 Early in DHF, a cascade of cytokines initiates a series of events that lead to a shift from a Th1-type response in mild DF to a Th2-type response resulting in severe DHF.5 Th1/Th2 polarisation is regulated by the induction of interleukin (IL)-10 and IL-12 by monocytes and monocyte-derived dendritic cells (DCs). Moreover, monocytes/macrophages express CD14, a documented DENV receptor; play a critical role in determining the balance of Th1/Th2 responses by modulating IL-12 production.

, 2005) Central memory cells and naive cells have high expressio

, 2005). Central memory cells and naive cells have high expression of CCR7 whereas effector memory cells have low expression of CCR7, the chemokine receptor for CCL21. It is likely that central memory cells are the

most responsive to CCL21 among all the subsets of CD8 T cells in our experiments. This is consistent with the increased speed during interstitial motility of central memory CD8 T cells compared to naive counterparts within intact lymph nodes in the absence BYL719 research buy of any antigen (Chtanova et al., 2009). Memory cells have increased surface levels of LFA1 compared to naive cells, which might contribute to higher responsiveness of central memory CD8 T cells to CCL21 co-adsorbed with ICAM1. We also observed that majority of CD45RO cells make contacts with the substratum, that are at least few microns in size, during CCL21-driven chemokinesis whereas majority of the CD45RA cells do not (Fig. 5b). These contacts are dynamic and discontinuous, similar to those observed previously in pre-activated T cells undergoing fast autonomous motility (Jacobelli et al., 2009). These contacts may also contribute to increased motility of CD45RO+ve cells. The novel findings reported in this study were critically SGI-1776 chemical structure dependent on integrating motility information with additional information from DIC, reflection and two fluorescence channels. In

the case of comparative analysis of CD45RA and CD45RO subsets, these were distinguished based on differential fluorescent dye labels. The fluorescence information allowed us to compare motility characteristics and reflection footprints of attachment simultaneously. This allowed us to delineate the motility and attachment tendencies of the subsets (Fig. 5b). Further delineation based on whether the cells within the subsets had shown contact footprint allowed us to observe that attachment promotes

motility (Fig. S12). In the case of LFA1 at the contact, its surface density could be related to motility characteristics and reflection footprints of attachment (Fig. 5c). We have brought together several existing approaches in building TIAM. The hybrid approach of edge detection followed by Hough transforms is a widely used approach for pattern recognition. Similarly the two-tier approach of linkage of neighboring cAMP objects in consecutive frames followed by temporal linkage of shorter segments is analogous to a recently introduced approach for single-particle tracking (Jaqaman et al., 2008). Put together, these approaches enable robust detection and tracking of cells. Accurate and comprehensive tracking is critical for developing motion models of cell motility and for characterizing heterogeneity in the motility behavior. Studying cellular heterogeneity has yielded better understanding of underlying mechanisms in other contexts (Altschuler and Wu, 2010). Our observation of an inverse relationship between the speed and turn angle of individual cells is a case in point (Fig.

Ravindran et al (2012) have described an LAMP method to detect L

Ravindran et al. (2012) have described an LAMP method to detect Las using primers for the 16S rDNA of the pathogen. However, for detection of HLB associated Las, LAMP has not been used widely so far. The availability of the full genome sequence of Las ( Duan et al., 2009) has enabled researchers to evaluate other regions of the bacterium that are more suitable for PCR-based detection technologies ( Morgan et al., 2012). We have developed a rapid, cost-effective, easy to operate, and field deployable technique to detect Las in psyllids. The method is very simple and can be routinely used effectively by citrus growers, extension

workers and home owners. It would be very useful to have quick and simple diagnostic tools to detect the pathogen in psyllid vectors in citrus growing regions of the world where facilities BYL719 order SGI-1776 datasheet are not available for expensive PCR testing. In addition, growers could afford regular monitoring of their groves. Extension workers and inspectors will have information that would enable them to alert a local testing laboratory if positive psyllids are detected. The first report of HLB in Louisiana was triggered by a report from a home owner who spotted a psyllid on

a “symptomatic tree” (Hummel and Ferrin, 2010). Utilizing the methodology and the instrumentation described in this work, we envision potential for implementing a wide surveillance program for detection of the pathogen. Rapid and reliable testing of a large number of psyllids combined with traditional methods of control, including targeted pesticide sprays to eliminate Las-positive psyllid sub-populations, will enable efficient and financially sustainable HLB management strategies. Psyllids maintained on HLB infected

plants in an insectary at the USDA ARS, United States Horticultural Research Laboratory, Fort Pierce were used for development of LAMP technology. Preserved psyllids stored in 95% ethanol were obtained from psyllid-infested regions of Florida, Brazil and Pakistan for testing in California. Las-free D. citri were obtained from a psyllid colony maintained at the quarantine facility located in the Dept. of Entomology, University of California Riverside (UCR), CA. Samples of the tomato psyllid, Bactericera cockerelli Clomifene maintained on tomato plants were obtained from Dept. of Entomology, UCR, CA. For the LAMP assay, crude ACP extracts were prepared as follows; 1–20 psyllids were removed from the collection tubes, the ethanol was air-dried on a piece of filter paper for 2–5 min and the psyllids were dropped into individually capped PCR tubes containing 100 μL of extraction buffer (20 mM Tris, pH 8.0 containing 2 mM EDTA and 1% TritonX100®) and heated in the Smart-DART™ unit for 10 min at 85 °C. The samples were centrifuged for 5 s in a micro-centrifuge and the clear supernatant was used for the LAMP assay. The tomato psyllids (B. cockerelli) carrying ‘Candidatus Liberibacter psyllaurous’ (synonym, Ca. L.

The equation at the current time step is expressed as equation(57

The equation at the current time step is expressed as equation(57) ξ¨1(t)ξ¨2(t)⋮ξ¨6+n(t)=[M+M(∞)]−1[f→(t)−M(∞)ξ¨1(t)ξ¨2(t)⋮ξ¨6+n(t)−Kξ1(t)ξ2(t)⋮ξ6+n(t)]where ξnξn is the modal displacement, the subscript n   is the mode number, subscripts 1–6 denote

rigid motion and subscripts 7 and higher denote flexible motion, and M(∞)M(∞) is the infinite frequency Palbociclib clinical trial added mass matrix. 4th order Adams–Bashforth–Moulton method is expressed as follows: equation(58) ξ̇′(t+Δt)=ξ̇(t)+Δt24[55ξ¨(t,ξ̇(t))−59ξ¨(t−Δt,ξ̇(t−Δt))+37ξ¨(t−2Δt,ξ̇(t−2Δt))−9ξ¨(t−3Δt,ξ̇(t−3Δt))] equation(59) ξ̇(t+Δt)=ξ̇(t)+Δt24[9ξ¨(t+Δt,ξ̇(t+Δt))+19ξ¨(t,ξ̇(t))−5ξ¨(t−Δt,ξ̇(t−Δt))+ξ¨(t−2Δt,ξ̇(t−2Δt))] Once the acceleration vector is obtained by solving Eq. (57), velocity and displacement are updated by 4th order Adams–Bashforth method in Eq. (58) as a predictor. Next, Eq. (57) is solved again to calculate the corrected acceleration vector, and the final values of velocity and displacement are recalculated by 4th order Adams–Moulton method in Eq. (59) as selleck chemical a corrector. Computation burden of GWM is not light even though it is a 2-D method. Slamming sections may experience water entry events with various initially submerged depths. Strictly,

for each water entry event, GWM solver should be run with the corresponding initial condition. Unfortunately, it leads to slow computation in time domain analysis. In order to reduce computation burden for GWM, a mapping scheme is used between GWM solutions with different initial conditions. A solution of GWM is independent of time histories of water entry motions because a gravity term is dropped off in the dynamic free surface condition. It means that the solution only depends on the initially submerged depth and the current water entry motion. For the mapping, the water entry problem is solved with the zero initial condition, which starts to enter the water from the zero submerged depth with a unit velocity. The solution of the problem is related to other slamming

events triclocarban with non-zero initial conditions. It is simple to relate two different initial value problems by applying offsets in the pile-up of the free surface. First, the water entry problem is solved for the section from the non-submerged condition to the fully-submerged condition. The solution of the problem is the pre-processed solution. In the solution, the submerged depth is decomposed into the penetration depth due to the relative vertical motion and the free surface elevation due to the water entry. When the section starts to enter the water from the depth of A, the wave elevation of W(A) can be found from the pre-processed solution. If the section penetrates the depth of C into the water, the corresponding solution should have the total submerged depth of C+W(C)−W(A). The modified penetration depth of X is obtained by solving the equation of X+W(X)=C+W(C)−W(A).

2), reef fish was the number one preference for more than 70% of

2), reef fish was the number one preference for more than 70% of respondents (Fig. 5). Chicken ranked similarly to tinned mTOR inhibitor fish and in the study households. A higher proportion of people preferred tilapia over fresh tuna, tinned fish and chicken, although fresh tuna ranked as the second preference for twice as many people as tilapia. Only five people ranked ‘salt-fish’ as their most preferred fish. The overall perception of tilapia was positive, with 98.3% of people surveyed familiar with the fish. Tilapia was described as a ‘good fish’ by 85% of respondents, with the majority saying this was because of its “good greasy taste” (Fig. 6). At the time of

the survey, with the exception of some small water storage areas, rudimentary backyard ponds and old drums, no tilapia was being farmed; all tilapia was being caught from nearby waterways (lakes, rivers and streams). Fourteen percent of respondents said that they had tried or had seen fish farming; in all cases this referred to tilapia, with the exception of one respondent who had experience in farming giant clams. Those who had tried growing tilapia in ponds reported a large range in pond size; on average approximately 4×4 m2 in area and 1–1.5 m in depth. Ponds were described as highly variable and opportunistic in design, taking advantage of natural depressions, large water drums or small creeks. Some

people did not feed their fish. For those GSI-IX that did, feeds were composed of white ants, kitchen scraps, coconut scrapings, rice, earthworms or mill run flour (in decreasing order of frequency mentioned). Ninety two percent of respondents, including men and women, expressed an interest in knowing more about, or undertaking, fish farming, primarily for household consumption. Sixteen percent (n=25) of respondents indicated that they were interested in watching the fish grow as a pastime, while two people indicated an interest in commercial production. One respondent oxyclozanide noted the value of farming tilapia for mosquito control purposes. When people who had previously attempted

to grow fish were asked why they had not continued with their ponds, they implied that they did not have sufficient knowledge to overcome any problems that they met, responding that they had found out about farming from friends and family that had very little knowledge or experience on fish farming. Some respondents had experienced their fish having being stolen. The lack of knowledge about husbandry practices, feeding and pond maintenance meant that farmers struggled to develop a productive farm and had become discouraged. The present study has provided insight into the fish and meat consumption patterns of peri-urban settlements in the vicinity of Auki and Honiara that have access to ‘wild’ sources of Mozambique tilapia to supplement their diets.

The S versicolor tree may reach up to 11 m in height, although i

The S. versicolor tree may reach up to 11 m in height, although in the western region of Mato Grosso do Sul most of the specimens are about 3–4 m in height. An outbreak of cattle mortality of unknown etiology, characterized by weakness, tremors, hind limbs incoordination and reluctance to move, was recorded on a farm in Água Clara, Mato Grosso do Sul, Brazil. The affected cattle were suspected

of feeding on S. versicolor, which was abundant on the property. The present study describes the epidemiology, clinical signs, necropsy and histopathological findings of spontaneous intoxication of cattle with S. versicolor and reproduces it experimentally. The outbreak was recorded on a farm with extensive livestock production in Água Clara city (20°37′05, 10″S, 52°36′01, 24″W, 303 m), click here eastern Mato Grosso do Sul State, Brazil (BR). Epidemiological data were provided by livestock handlers interviewed during visits to the farm. Two animals were necropsied, one 12 h postmortem and the other after

clinical examination followed by euthanasia. Organ fragments were collected, fixed in 10% formaldehyde solution, subjected to routine methods and stained with hematoxylin and eosin (HE) for histological examination. The paddocks where the dead animals were found contained not only forage grass (Brachiaria decumbens and Brachiaria brizantha) but also invasive toxic plants, which were collected EPZ5676 solubility dmso and submitted for botanical identification at the Botany Laboratory and kept in the Herbarium (CGMS) of the Federal University of Mato Grosso do Sul (UFMS). The species identified were S. versicolor (CGMS – 34897), Senna occidentalis, Senna obtusifolia and Crotalaria mucronata. Pasture was evaluated in terms of forage supply, i.e., areas where pasture height was lower than 10 cm were considered areas with depleted forage availability, whereas areas where pasture height was 20–40 cm were considered to contain a good forage supply (Sbrissia, 2004). To experimentally reproduce intoxication, green leaves without Inositol monophosphatase 1 stems of S. versicolor were collected in the paddock where the cattle died during the outbreak. The leaves were stored in

plastic bags and frozen at −5 °C. Each portion of leaves fed to the cattle was removed from the freezer 24 h before administration. Two experiments were conducted (Table 1), using four crossbred male calves aged 8–12 months. The animals weighed 121, 110, 130 and 185 kg (calves 1, 2, 3, and 4, respectively) and were held in individual stalls supplied with water, alfalfa hay and feed. The experiments were approved by the Animal Ethics Committee (CEUA) of UFMS (protocol number 400/2012). In experiment 1, three animals (calf 1, 2 and 3) were administered single leaf doses of 15 g/kg, 5 g/kg and 2.5 g/kg, respectively, to determine the toxic dosage of the plant. In calf 1, the leaves were given by rumen cannula, and for calves 2 and 3, they were administered orally.