Influência da qualidade micotoxicológica e nutricional de híbridos de milho no custo da ração de frangos de corte / Influence of mycotoxicological and nutritional quality of maize hybrids on broiler chickens feed cost

Objetivou-se avaliar as variáveis micotoxicológicas e nutricionais de híbridos de milho com diferentes características que influenciam no custo da ração para frangos de corte. Foram avaliados 26 híbridos de milho geneticamente modificados nas safrinhas de 2016 e 2017, com diferentes germoplasmas, textura de endosperma e duração do ciclo. Nos híbridos, foram avaliados grãos avariados, fumonisinas (B1+B2) (FUM), aflatoxinas (B1+B2+G1+G2) (AFLA), zearalenona (ZEA), deoxinivalenol (DON), umidade, proteína bruta (PB), energia metabolizável aparente corrigida para balanço de nitrogênio (EMAn), aminoácidos digestíveis para aves (lisina, metionina, cistina e treonina) e o respectivo custo da ração inicial para frangos de corte, que foi calculada pelo custo mínimo. A prevalência de FUM, AFLA, ZEA e DON foi de 90, 17, 33 e 0%, com médias de 3067, 1, 38 e 0µg/kg nos dois anos, respectivamente. A média de EMAn e PB foi de 3264kcal/kg e 8,02%, respectivamente, e diferiu (P<0,05) nos dois anos. O custo da ração foi influenciado significativamente (P<0,05) por FUM, PB, EMAn nos dois anos. Híbridos com tecnologia Viptera apresentam menor concentração por FUM e menor custo da ração. Híbridos de ciclo precoce têm menor concentração de FUM, maiores percentuais de PB e de aminoácidos digestíveis e menor custo da ração.(AU)

The objective of this study was to evaluate the mycotoxicological and nutritional variables of maize hybrids with different characteristics that influence the broiler chicken's feed costs. In 2016 and 2017 winter crops, 26 genetically modified hybrids of maize with different germplasm, endosperm texture and cycle duration were evaluated. The analyzed variables were damaged grains, fumonisins (B 1 +B 2 ) (FUM), aflatoxins (B 1 +B 2 +G 1 +G 2 ) (AFLA), zearalenone (ZEA), deoxynivalenol (DON), moisture, crude protein (CP), apparent metabolizable energy corrected for nitrogen balance (AMEn), digestible amino acids for poultry (lysine, methionine, cystine and threonine) and the respective cost of the initial feed for broiler chickens calculated at the minimum cost. The prevalence of FUM, AFLA, ZEA and DON was 90, 17, 33 and 0%, with means of 3067, 1, 38 and 0µg/kg in the two years, respectively. The mean of AMEn and CP was 3264kcal/kg and 8.02%, respectively, and differed (P< 0.05) in the two years. The feed cost was significantly influenced (P<0.05) by FUM, PB, AMEn in two years. Hybrids with Viptera technology show lower concentration per FUM and lower feed cost. Early cycle hybrids have lower concentrations of FUM, higher percentages of CP and digestible amino acids, and lower feed costs.(AU)

Seasonal variability of the essential oil of Hesperozygis ringens (Benth.) Epling.

This study was developed to evaluate the effect of seasonality on the yield and chemical composition of the essential oil (EO) of Hesperozygis ringens (Benth.) Epling, a native species from the Brazilian Pampa. Leaves were collected from four specimens of a single population in each of the four seasons for a year and were extracted in triplicate by hydro-distillation for 2 hours. The yield of EO (% w/w) was calculated on fresh weight basis (FWB), and the 16 oil samples were analyzed by gas chromatography coupled to mass spectrometry (GC-MS) and gas chromatography with flame ionization detector (GC-FID). Hierarchical Cluster Analysis (HCA) and Principal Component Analysis (PCA) were used as statistical tools to evaluate differences in chemical composition. The highest yields were obtained in autumn, spring and summer (2.32-4.38%), while the lowest yields were detected in winter, ranging from 1.15 to 1.91%. Oxygenated monoterpenoids were the predominant class of chemical constituents in the EO obtained in all seasons, showing the highest contents in autumn and summer, and pulegone was identified as a major compound, whose contents varied between 54.13 and 81.17%. The EO samples were divided into three chemical groups by HCA and PCA and were assigned to the same group, except for the three samples gathered in winter. The results showed a seasonal influence on the yield and chemical composition of the EO.

Seasonal variability of the essential oil of Hesperozygis ringens (Benth.) Epling. / Variabilidade sazonal do óleo essencial de Hesperozygis ringens (Benth.) Epling.

Abstract This study was developed to evaluate the effect of seasonality on the yield and chemical composition of the essential oil (EO) of Hesperozygis ringens (Benth.) Epling, a native species from the Brazilian Pampa. Leaves were collected from four specimens of a single population in each of the four seasons for a year and were extracted in triplicate by hydro-distillation for 2 hours. The yield of EO (% w/w) was calculated on fresh weight basis (FWB), and the 16 oil samples were analyzed by gas chromatography coupled to mass spectrometry (GC-MS) and gas chromatography with flame ionization detector (GC-FID). Hierarchical Cluster Analysis (HCA) and Principal Component Analysis (PCA) were used as statistical tools to evaluate differences in chemical composition. The highest yields were obtained in autumn, spring and summer (2.32-4.38%), while the lowest yields were detected in winter, ranging from 1.15 to 1.91%. Oxygenated monoterpenoids were the predominant class of chemical constituents in the EO obtained in all seasons, showing the highest contents in autumn and summer, and pulegone was identified as a major compound, whose contents varied between 54.13 and 81.17%. The EO samples were divided into three chemical groups by HCA and PCA and were assigned to the same group, except for the three samples gathered in winter. The results showed a seasonal influence on the yield and chemical composition of the EO.

Resumo Este estudo foi desenvolvido a fim de avaliar o efeito da sazonalidade no rendimento e composição química do óleo essencial (OE) de Hesperozygis ringens (Benth.) Epling., uma espécie nativa do Pampa brasileiro. Folhas foram coletadas de quatro indivíduos de uma mesma população, em cada uma das quatro estações de um ano, e foram extraídas em triplicada por hidrodestilação durante 2 horas. O rendimento do OE (% m/m) foi calculado considerando a base fresca (BF) e as 16 amostras de óleo foram analisadas por cromatografia gasosa acoplada à espectrometria de massas (CG-EM) e cromatografia gasosa com detector de ionização de chamas (CG-DIC). Análise Hierárquica de Cluster (AHC) e Análise de Componentes Principais (ACP) foram utilizadas como ferramentas estatísticas para avaliar as diferenças na composição química. Os maiores rendimentos foram obtidos no outono, primavera e verão (2,32-4,38%), enquanto que os menores foram detectados no inverno, variando de 1,15 até 1,91%. Os monoterpenoides oxigenados foram a classe predominante dos constituintes do OE obtido em todas as estações, apresentando os maiores teores no outono e no verão, e a pulegona foi identificada como o constituinte majoritário, cujos teores variaram entre 54,13 e 81,17%. As amostras de OE foram divididas em três grupos químicos por AHC e ACP e foram classificadas no mesmo grupo, com exceção de três amostras coletadas no inverno. Os resultados demonstraram influência sazonal no rendimento e na composição química dos OE.

Involvement of oxidative stress in subacute toxicity induced by fumonisin B1 in broiler chicks.

Fumonisin B1 (FB1) is a mycotoxin produced by Fusarium spp. It has been reported to be a potential cause of liver cancer in rats and esophageal cancer in humans. The underlying mechanisms of FB1 toxicity are thought to be related to the inhibition of ceramide synthase, causing an accumulation of sphingosine (SO) and sphinganine (SA), which in turn may cause tissue functional impairment and the development of oxidative stress. Therefore, in this study, we investigate the effects of an FB1-contaminated diet on markers of oxidative stress in chick liver. A total of 24 male broiler chicks (Cobb 500) were fed a standard control diet or a diet contaminated with FB1 (100mg/kg) for 21 days, starting on postnatal day one. The feed and animals were weighed on days 0, 7, 14 and 21 to estimate the feed conversion ratio, and at 21 days, the liver weight and liver relative weight were determined. At the end of the experiment, samples of blood and liver were collected. The blood was used to quantify the SA/SO ratio, and the liver was used to determine the activity of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST); ascorbic acid levels (VitC), non-protein thiol (NPSH) levels and TBARS content were also determined. The FB1 diet increased the liver weight, liver relative weight, feed conversion and SA/SO ratio. Furthermore, hepatic TBARS levels, Vit C content and CAT activity were also increased. Conversely, the activities of SOD, GST and NPSH levels, in the liver were not altered by the mycotoxin-contaminated diet. In summary, we showed that subacute exposure of broiler chicks to FB1 induced liver oxidative stress concomitantly with SA/SO accumulation.

Sedative and anesthetic activities of the essential oils of Hyptis mutabilis (Rich.) Briq. and their isolated components in silver catfish (Rhamdia quelen).

This study evaluated the sedative and anesthetic effects of the essential oils (EO) of Hyptis mutabilis (Rich.) Briq. and their isolated components on silver catfish (Rhamdia quelen). Quantitative chemical differences between the EOs obtained from leaves and inflorescences were verified, and a new chemotype rich in globulol was described. Although there were no significant differences in the time of induction for sedation and anesthesia between the EOs, only the leaf EO at 344 mg/L anesthetized all fish without side effects. Fractionation of the leaf EO was carried out by column chromatography. The isolated compounds [(+)-1-terpinen-4-ol and (-)-globulol] showed different activity from that detected for the leaf EO in proportional concentrations and similar sedation to a eugenol control at 10 mg/L. However, fish exposed to 1-terpinen-4-ol (3 and 10 mg/L) did not remain sedated for 30 min. Anesthesia was obtained with 83-190 mg/L globulol, but animals showed loss of mucus during induction and mortality at these concentrations. Synergism of the depressor effects was detected with the association of globulol and benzodiazepine (BDZ), compared with either drug alone. Fish exposed to BDZ or globulol+BDZ association showed faster recovery from anesthesia in water containing flumazenil, but the same did not occur with globulol. In conclusion, the use of globulol in aquaculture procedures should be considered only at sedative concentrations of 10 and 20 mg/L, and its mechanism of action seems not to involve the GABAA-BDZ system.

Sedative and anesthetic activities of the essential oils of Hyptis mutabilis (Rich.) Briq. and their isolated components in silver catfish (Rhamdia quelen)

This study evaluated the sedative and anesthetic effects of the essential oils (EO) of Hyptis mutabilis (Rich.) Briq. and their isolated components on silver catfish (Rhamdia quelen). Quantitative chemical differences between the EOs obtained from leaves and inflorescences were verified, and a new chemotype rich in globulol was described. Although there were no significant differences in the time of induction for sedation and anesthesia between the EOs, only the leaf EO at 344 mg/L anesthetized all fish without side effects. Fractionation of the leaf EO was carried out by column chromatography. The isolated compounds [(+)-1-terpinen-4-ol and (-)-globulol] showed different activity from that detected for the leaf EO in proportional concentrations and similar sedation to a eugenol control at 10 mg/L. However, fish exposed to 1-terpinen-4-ol (3 and 10 mg/L) did not remain sedated for 30 min. Anesthesia was obtained with 83-190 mg/L globulol, but animals showed loss of mucus during induction and mortality at these concentrations. Synergism of the depressor effects was detected with the association of globulol and benzodiazepine (BDZ), compared with either drug alone. Fish exposed to BDZ or globulol+BDZ association showed faster recovery from anesthesia in water containing flumazenil, but the same did not occur with globulol. In conclusion, the use of globulol in aquaculture procedures should be considered only at sedative concentrations of 10 and 20 mg/L, and its mechanism of action seems not to involve the GABAA-BDZ system.

Effects of aflatoxins on performance and exocrine pancreas of broiler chickens.

The aim of this research was to evaluate, on a weekly basis, the effects of aflatoxins on the activity of digestive enzymes (alpha-amylase, lipase, and trypsin) in the pancreas as well as on the performance and histology of pancreas in broiler chickens over the course of 42 days. One thousand and eighty 1-day-old male Cobb broilers were divided into four treatments with 18 replicates and 15 birds per replicate (i.e., 270 broilers per treatment). Treatments were established according to the amount of aflatoxins added to the diet, as follows: T1 = 0 mg of aflatoxins per kilogram of feed (mg/kg); T2 = 0.7 mg/kg; T3 = 1.7 mg/kg; and T4 = 2.8 mg/kg. Pancreas sample collection was performed from one bird out of each replicate at 7, 14, 21, 28, 35, and 42 days of experiment, which yielded a total of 18 samples per treatment on each collection. Each sample was homogenized in distilled water, frozen in liquid nitrogen, lyophilized, and stored at -20 C until analysis. Performance parameters (body weight, feed consumption, and feed conversion rate) were measured at 21, 35, and 42 days of experiment. At the end of the experiment (42 days), six birds from each treatment were randomly chosen for histologic evaluation of the pancreas. The presence of aflatoxins in the diet induced a negative effect on all performance parameters. The pancreatic activity of lipase and alpha-amylase were significantly increased in treatments T3 and T4, while the specific activity of trypsin was only affected during treatment T4. In addition, several histologic changes were observed in the pancreas of birds receiving aflatoxin-contaminated feed. Aflatoxins present in the feed determined an increase in the activity of pancreatic enzymes in broilers, affecting the digestibility of the diet, thereby leading to losses in performance and productivity.

Composition and evaluation of the antimicrobial activity of the essential oil of Senecio selloi Spreng DC / Composição e avaliação da atividade antimicrobiana do óleo essencial de Senecio selloi Spreng DC

The essential oil of the aerial parts of Senecio selloi Spreng. DC. was extracted by hydrodistillation and analyzed by GC/MS. Nineteen compounds were identified, representing 99.9% of the total. The main compounds were found to be sesquiterpene hydrocarbons (71.3%), most of them with a bisabolane skeleton (59.4%). The major constituent was α-zingiberene (54%), followed by monoterpene α-isolimonene (16%). The essential oil was also tested against two Gram-positive and two Gram-negative bacterial species, three yeasts, and an algae. From the strains assayed, only Bacillus subtilis ATCC 6633 showed susceptibility (MIC and MBC = 4400 µg/mL) to the essential oil.

O óleo essencial das partes aéreas de Senecio selloi Spreng DC. foi extraído por hidrodestilação e analisado por CG/EM. Dezenove constituintes foram identificados, representando 99,9% do total. Os principais compostos fornecidos foram sesquiterpenos hidrocarbonetos (71,3%), a maioria destes com esqueleto bisabolano (59,4%). O constituinte majoritário foi a-zingibereno (54%), seguido do monoterpeno a-isolimoneno (16%). O óleo essencial foi testado contra duas cepas Gram-positivas e duas Gram-negativas, três fungos e uma alga. De todas as linhagens testadas somente Bacillus subtilis ATCC 6633 mostrou suscetibilidade (CIM e CBM = 4400 µg/mL) para o óleo essencial.

Investigação do gene p53 de frangos expostos às aflatoxinas / Investigation of p53 gene in poultries exposed to aflatoxins

Identificou-se o efeito das aflatoxinas (AFs) sobre o gene p53 de frangos de corte, de linhagem comercial, separados em: grupo experimental, tratado (GT) com ração comercial contendo 2,8ppm de AFs totais durante 21 dias consecutivos, e grupo-controle (GC), sem exposição às AFs. Macroscopicamente, as alterações caracterizaram-se por hepatomegalia e aspecto pálido-amarelado com alguns focos hemorrágicos e, histologicamente, por desarranjo trabecular, pleomorfismo hepatocítico com cariomegalia, degeneração vacuolar intracitoplasmática, necrose com infiltração linfocítica e hiperplasia de ductos biliares. A PCR com os primers GSPT53c-1 com base no gene candidato a p53 (GenBank XM_424937.2) gerou um produto de aproximadamente 350 pares de base. O amplicon sequenciado a partir do DNA dos frangos do GT não apresentou mutação ou deleção, assim como padrão de bandas do PCR-RFLP não foi distinto entre ambos os grupos experimentais e a sequência depositada no banco de genes. Os resultados sugerem que não ocorreu transversão devido à exposição às AFs no fragmento amplificado. Conclui-se que a PCR-RFLP e o sequenciamento do produto da PCR não são ferramentas apropriadas para diagnóstico da exposição de frangos às AFs nas condições experimentais empregadas.

To identify the effects of aflatoxins (AFs), Cobb lineage poultry were separated in an experimental group in which they were treated with commercial ration containing 2.8ppm of total AFs during 21 days (TG) and a control group without AFs exposure (CG). In the liver of poultries exposed to AFs, alterations were microscopically observed, which were characterized by hepatomegaly, a pale yellowish aspect with some hemorrhagic spots, and histologically a trabecullar disarranging pleomorphic hepatocytes with cariomegaly, intracytoplasmatic vacuolar degeneration, necrosis, lymphocytic infiltration and hyperplasia of biliary ducts. The PCR with GSPT53c-1 primers based on p53 candidate gen (GenBank XM_424937.2) generated a product of approximately 350 base pairs. The sequenced amplicon obtained from the DNA of treated poultry did not display any mutation or deletion, and the PCR- RFLP bands patterns were also not distinct in both experimental groups. The results indicated that transversion did not occur in the fragment amplified due to AFs exposure. As a consequence of results obtained with p53 gene (NM_205264.1) we concluded that PCR-RFLP and sequencing of PCR product are not appropriate diagnostic tools for the detection of poultry exposure to AFs, at least in the experimental conditions performed.

Individual and combined effects of Salmonella typhimurium lipopolysaccharide and fumonisin B1 in broiler chickens.

The aim of this research was to evaluate the individual and combined effects of Salmonella typhimurium lipopolysaccharide (sLPS) and fumonisin B(1) (FB) on performance, relative weight of liver, biological parameters, and histological evaluation of several tissues from four hundred thirty-two 1-d-old male broiler chickens divided into 9 treatments according to the dose of FB (0, 100, or 200 mg/kg, from d 1 to d 28) and sLPS (0, 250, or 500 µg/application per bird, every other day, from d 15 to 27) administered. At the end of the experiment (28 d), significant effects caused by sLPS, FB, and the interaction of sLPS × FB were observed on several parameters. Histopathological evaluations showed significant lesions in liver and kidney caused by sLPS, FB, and their association. According to these results, both sLPS and FB (isolated or in association) cause significant effects on performance and biological parameters of broilers at 28 d of age.

Participation of the GABAergic system in the anesthetic effect of Lippia alba (Mill.) N.E. Brown essential oil

The objective of this study was to identify the possible involvement of the GABAergic system in the anesthetic effect of Lippia alba essential oil (EO). We propose a new animal model using silver catfish (Rhamdia quelen) exposed to an anesthetic bath to study the mechanism of action of EO. To observe the induction and potentiation of the anesthetic effect of EO, juvenile silver catfish (9.30 ± 1.85 g; 10.15 ± 0.95 cm; N = 6) were exposed to various concentrations of L. alba EO in the presence or absence of diazepam [an agonist of high-affinity binding sites for benzodiazepinic (BDZ) sites coupled to the GABA A receptor complex]. In another experiment, fish (N = 6) were initially anesthetized with the EO and then transferred to an anesthetic-free aquarium containing flumazenil (a selective antagonist of binding sites for BDZ coupled to the GABA A receptor complex) or water to assess recovery time from the anesthesia. In this case, flumazenil was used to observe the involvement of the GABA-BDZ receptor in the EO mechanism of action. The results showed that diazepam potentiates the anesthetic effect of EO at all concentrations tested. Fish exposed to diazepam and EO showed faster recovery from anesthesia when flumazenil was added to the recovery bath (12.0 ± 0.3 and 7.2 ± 0.7, respectively) than those exposed to water (9.2 ± 0.2 and 3.5 ± 0.3, respectively). In conclusion, the results demonstrated the involvement of the GABAergic system in the anesthetic effect of L. alba EO on silver catfish.

Participation of the GABAergic system in the anesthetic effect of Lippia alba (Mill.) N.E. Brown essential oil.

The objective of this study was to identify the possible involvement of the GABAergic system in the anesthetic effect of Lippia alba essential oil (EO). We propose a new animal model using silver catfish (Rhamdia quelen) exposed to an anesthetic bath to study the mechanism of action of EO. To observe the induction and potentiation of the anesthetic effect of EO, juvenile silver catfish (9.30 ± 1.85 g; 10.15 ± 0.95 cm; N = 6) were exposed to various concentrations of L. alba EO in the presence or absence of diazepam [an agonist of high-affinity binding sites for benzodiazepinic (BDZ) sites coupled to the GABA A receptor complex]. In another experiment, fish (N = 6) were initially anesthetized with the EO and then transferred to an anesthetic-free aquarium containing flumazenil (a selective antagonist of binding sites for BDZ coupled to the GABA A receptor complex) or water to assess recovery time from the anesthesia. In this case, flumazenil was used to observe the involvement of the GABA-BDZ receptor in the EO mechanism of action. The results showed that diazepam potentiates the anesthetic effect of EO at all concentrations tested. Fish exposed to diazepam and EO showed faster recovery from anesthesia when flumazenil was added to the recovery bath (12.0 ± 0.3 and 7.2 ± 0.7, respectively) than those exposed to water (9.2 ± 0.2 and 3.5 ± 0.3, respectively). In conclusion, the results demonstrated the involvement of the GABAergic system in the anesthetic effect of L. alba EO on silver catfish.

Immunoassay based on monoclonal antibodies versus LC-MS: deoxynivalenol in wheat and flour in Southern Brazil.

An indirect competitive enzyme-linked immunosorbent assay (ELISA) method using a monoclonal antibody for deoxynivalenol (DON) detection in wheat and flour was standardised and validated (detection limit = 177.1 µg kg(-1)) and its performance was compared with LC-MS, quantification limit =140 µg kg(-1)). DON recovery ranged from 88.7% to 122.6% for wheat grain and from 70.6% to 139.3% for flour. Among the 38 wheat samples evaluated, DON was detected in 29 samples (76.3%) by ic-ELISA (281.6-12 291.4 µg kg(-1)) and in 22 samples (57.9%) by LC-MS (155.3-9906.9 µg kg(-1)). The 0.93 correlation coefficient between ic-ELISA and LC-MS data in 19 positive DON wheat samples demonstrated the reliability and efficiency of ic-ELISA. Results indicated that standardised ic-ELISA was suitable for DON screening in wheat samples and the need for continuous monitoring of mycotoxin levels in foodstuffs.

Toxicokinetics and toxicological effects of single oral dose of fumonisin B1 containing Fusarium verticillioides culture material in weaned piglets.

Toxicokinetics and the toxicological effects of culture material containing fumonisin B(1) (FB(1)) were studied in male weaned piglets by clinical, pathological, biochemical and sphingolipid analyses. The animals received a single oral dose of 5 mg FB(1)/kg of body weight, obtained from Fusarium verticillioides culture material. FB(1) was detected by HPLC in plasma collected at 1-h intervals up to 6h and at 12-h intervals up to 96 h. FB(1) eliminated in feces and urine was quantified over a 96-h period and in liver samples collected 96 h post-intoxication. Blood samples were obtained at the beginning and end of the experiment to determine serum enzyme activity, total bilirubin, cholesterol, sphinganine (Sa), sphingosine (So) and the Sa/So ratio. FB(1) was detected in plasma between 30 min and 36 h after administration. The highest concentration of FB(1) was observed after 2 h, with a mean concentration of 282 microg/ml. Only 0.93% of the total FB(1) was detected in urine between 75 min and 41 h after administration, the highest mean concentration (561 microg/ml) was observed during the interval after 8 at 24 h. Approximately 76.5% of FB(1) was detected in feces eliminated between 8 and 84 h after administration, with the highest levels observed between 8 and 24 h. Considering the biochemical parameters, a significant increase only occurred in cholesterol, alkaline phosphatase and aspartate aminotransferase activities. In plasma and urine, the highest Sa and Sa/So ratios were obtained at 12 and 48 h, respectively.

Alimentação de leitoas pré-púberes com dietas contendo zearalenona / Feeding of pre-pubertal gilts with diets containing zearalenone

O desempenho, o peso de alguns órgãos e a morfologia vulvar de leitoas pré-púberes, alimentadas por 28 dias com dietas contendo zearalenona, foram avaliados. O delineamento experimental utilizado foi inteiramente ao acaso, com dois tratamentos, dieta controle (DC) e dieta controle + 2mg kg-1 de zearalenona (DZ), e seis repetições cada. Não houve diferença (P>0,05) entre os tratamentos para consumo médio diário de ração (1,24 x 1,19kg), ganho médio diário de peso (0,68 x 0,71kg), conversão alimentar (1,86 x 1,71) e peso vivo (PV); (30,9 x 30,4kg). A zearalenona não alterou (P>0,05) os pesos absoluto e relativo do coração (137 x 141g e 0,45 x 0,45 por cento PV), fígado (699 x 699g e 2,31 x 2,26 por centoPV), rins (47 x 49g e 0,15 x 0,16 por centoPV) e baço (166 x 171g e 0,55 x 0,55 por centoPV). Houve aumento (P<0,05) no comprimento (17 x 27cm) e no peso (23 x 157g e 0,07 x 0,51 por centoPV) do trato reprodutivo das leitoas do grupo DZ. O volume vulvar ao final do período foi 820 por cento maior (P<0,05) nos animais alimentados com zearalenona (941 x 8658mm³/kgPV0,6). Os resultados indicam que em suínos a zearalenona e seus metabólitos possuem atividade estrogênica, mas não interferem no desempenho dos animais.

The performance, the weights of some organs, and the vulvae morphology in pre-pubertal gilts fed diets containing zearalenone were evaluated during 28 days. The experimental design was completely randomized with two treatments (control diet, ZD - control diet + 2mg kg-1 of zearalenone) and six replications of each were done. No differences (P>0.05) between treatments for daily feed intake (1.24 x 1.19kg), average daily gain (0.68 x 0.71kg), feed conversion ratio (1.86 x 1.71), and live weight (30.9 x 30.4kg) were observed. Zearalenone did not change (P>0.05) the absolute and relative weights of heart (137 x 141g and 0.45 x 0.45 percentBW), liver (699 x 699g and 2.31 x 2.26 percentBW), kidneys (47 x 49g and 0.15 x 0.16 percentBW), and spleen (166 x 171g and 0.55 x 0.55 percent BW). However, zearalenone increased (P<0.05) the length (17 x 27cm) and weight (23 x 157g and 0.07 x 0.51 percentBW) of the reproductive tract. The final vulvae volume was 820 percent larger (P<0.05) in gilts fed diets containing zearalenone than those fed control diet (941 x 8658mm³/kgBW0.6). Results suggested that zearalenone and its metabolites have an estrogenic activity in pigs without changing the animal performance.

Otimização de método analítico para determinação de aflatoxinas em rações de caprinos e ocorrência no estado da Bahia / Optimization of analytical method for aflatoxins determination in goats feedstuffs and their occurrence in the state of Bahia

The optimization of the thin layer chromatography (TLC) method to determine aflatoxins in feedstuffs and the evaluation of their occurrence in feedstuffs for milk-yielding does in the state of Bahia were studied. Eighty feedstuff samples were collected from five farms, located in the Recôncavo Baiano, from November 2000 to August 2002. The samples were analyzed using TLC modified method. In this study, the detection and quantification limits were 5 and 8m g/kg, respectively. The percentage of average recoveries obtained for AFB1, AFB2, AFG1, and AFG2 were 81.0; 97.2; 89.3; and 90.3 percent, respectively; and the coefficient of variation ranged from 0.83 to 4.1 percent. The results revealed that the optimization of TLC methodology is efficient to analyze aflatoxins in feedstuffs. The presence of these aflatoxins was not detected in any of the analyzed samples, demonstrating good quality of those products, regarding the contamination by these toxins.

Performance of turkey poults fed different doses of aflatoxins in the diet.

This work was conduced to determine the performance parameters of initial-phase turkey poults fed 7 different doses of aflatoxins in the diet. Three hundred thirty-six 1-d-old male turkey poults were used in this research. Turkeys were divided into 7 treatments according to aflatoxin doses (T1 = control; T2 = 20 ppb aflatoxins; T3 = 50 ppb; T4 = 100 ppb; T5 = 200 ppb; T6 = 500 ppb; T7 = 1,000 ppb). Birds were killed in 2 periods: half of them after 21 d of experiment and the remaining birds after 42 d of experiment. In both periods, the evaluated parameters were as follows: feed consumption, BW, relative weights of organs (liver, gizzard, heart, and bursa of Fabricius) and meat (breast and thighs), and clinical biochemistry parameters (total plasmatic proteins, albumin, uric acid, cholesterol, alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase). At 21 d of experiment, both feed consumption and BW were significantly affected by the aflatoxins present in the diet. Nevertheless, gizzard relative weight, total plasmatic proteins, and cholesterol levels were also affected. At the 42-d evaluation, besides feed consumption and BW, gizzard and liver relative weights and cholesterol levels were also affected by the presence of aflatoxins in the diet. Turkey poults are very sensitive to aflatoxin poisoning, because they are at least 3 to 6 times more sensitive to these contaminants than broilers.

Determination of ochratoxin A in green coffee by immunoaffinity column cleanup and liquid chomatography: collaborative study.

A collaborative study was conducted to evaluate a method using immunoaffinity column cleanup with liquid chromatography (LC) for the determination of ochratoxin A (OTA) in green coffee at levels that could be included in possible future regulations of the European Union. The test portion was extracted with methanol-3% aqueous sodium hydrogen carbonate solution (50 + 50, v/v). The extract was filtered, and the filtrate was diluted with phosphate-buffered saline and applied to an immunoaffinity column containing antibodies specific for OTA. After washing, the toxin was eluted from the column with methanol and quantified by LC with fluorescence detection. Pairs of 4 homogeneous noncontaminated and naturally contaminated materials (mean levels of < 0.12, 2.44, 5.15, and 13.46 ng/g) and blank samples (< 0.12 ng/g) for spiking were sent to 20 participant laboratories from 8 countries. The materials were analyzed according to the method description and all difficulties encountered in the analysis were reported. Statistical analysis was carried out according to the Harmonized Protocol of the International Union of Pure and Applied Chemistry. The relative standard deviation for repeatability (RSDr) ranged from 7.42 to 20.94%, and the relative standard deviation for reproducibility (RSDR) ranged from 16.34 to 29.17%. The method showed acceptable within-laboratory and between-laboratories precision for green coffee materials, as evidenced by HorRat values of < or = 0.85, at the studied range, for spiked and naturally contaminated materials. The mean recovery was 92.8% for green coffee material spiked with OTA at a level of 4.82 ng/g.

Toxicological effects of chronic low doses of aflatoxin B(1) and fumonisin B(1)-containing Fusarium moniliforme culture material in weaned piglets.

The effects of chronic oral exposure (28 days) to aflatoxin B(1) (AFB(1)) and fumonisin B(1) (FB(1)) were studied in weaned piglets. Six experimental groups, each comprising two neutered males and two females, were fed ad libitum with rations containing: (A) 0 mg of FB(1) and 0 mg of AFB(1)/kg of feed (control); (B) 10 mg of FB(1)/kg of feed; (C) 30 mg of FB(1)/kg of feed; (D) 50 microg of AFB(1)/kg of feed; (E) 10 mg of FB(1) plus 50 microg of AFB(1)/kg of feed; (F) 30 mg of FB(1) plus 50 microg of AFB(1)/kg of feed. The animals were inspected twice daily and their body weight and feed consumption were recorded weekly and daily, respectively. Samples of feces and urine were collected 24 h after the start of the experiment, to check for fumonisin residues by HPLC analysis. Blood samples were drawn at the start of the experiment and after 28 days for quantification of hematological and biochemical parameters. Necropsies were performed after 28 days; at necropsy, the organs were weighed, inspected macroscopically and processed for histopathological and toxicological analyses. All piglets from groups C and F presented typical signs of pulmonary edema, with reduced feed consumption and body weight gain as well as pathological alterations. FB(1) was detected in feces and urine at 24 h of intoxication and in liver after 28 days of intoxication. Increases were detected regarding the following hematological and biochemical parameters in animals from treatments C and F: erythrocyte number; hematocrit; total bilirubin; total protein; activity of serum alkaline phosphatase, aspartate aminotransferase, and alanine aminotransferase. Cholesterol levels were significantly aumented only in animals from groups C and F, whereas albumin concentrations increased in groups C, F, B and E. The average organ/body weight ratio of piglets (hearth, liver and lung) were significantly greater in groups C and F. The only joint effects of FB(1) and AFB(1) detected (group F) were a decrease in feed consumption during the last week of intoxication and in feed conversion throughout the 28 days of intoxication. Chronic intoxication of piglets with AFB(1) and FB(1) leads to important losses of productivity.

Robotic automated clean-up for detection of fumonisins B1 and B2 in corn and corn-based feed by high-performance liquid chromatography.

A high-performance liquid chromatography (HPLC) system with fluorescence detection and an automated on-line solid-phase extraction procedure for fumonisins B1 and B2 in corn and corn-based products is described. Different amounts of strong anion-exchange, C18 and end-capped C18 (C(18 ec)) silicas were tested for sample clean-up. Various HPLC parameters were analyzed. The best methodology was found to be extraction with acetonitrile-water and clean up on C(18 ec) disposable extraction cartridges. The system has the advantage of running in an unattended mode of operation and allows processing of 40 samples without system refuel, performing clean-up, o-phthaldialdehyde derivatization, injection and fumonisin detection by fluorescence detection linked to a computer integrator for automated data processing. Recoveries were performed with corn and corn-based feed samples (n=3) spiked with 0.1, 0.5, 1.0, 5.0 and 10 microg/g. Average recoveries for corn and corn-based feed were, respectively, 92.6 and 88.3% with relative standard deviations (RSDs) of 5.04 and 6.22%, for fumonisin B1 and 91.2 and 89.0% with RSDs of 5.84 and 7.88% for fumonisin B2. Detection limits (S/N=3) for corn and corn-based feed were approximately 0.03 microg/g for fumonisin B1 and 0.05 microg/g for fumonisin B2