New Nanostructured Silica Adjuvant (SBA-15) Employed to Produce Antivenom in Young Sheep Using Crotalus durissus terrificus and Apis mellifera Venoms Detoxified by Cobalt-60

Equine antivenom is considered the only treatment for animal-generated envenomations, but it is costly. The study aimed to produce Apis mellifera (Africanized honeybee) and Crotalus durissus terrificus (C.d.t.) antivenoms using nanostructured silica (SBA-15) as adjuvant and cobalt-60 (60Co)-detoxified venoms utilizing young sheep. Natural and 60Co-irradiated venoms were employed in four different hyperimmunization protocols. Thus, 8 groups of 60- to 90-d-old sheep were hyperimmunized, enzyme-linked immunosorbent assay (ELISA) serum titers collected every 14 d were assessed clinically daily, and individual weight were measured, until d 84. Incomplete Freund's (IFA) and nanostructured silica (SBA15) adjuvants were compared. The lethal dose (LD50) for both venoms was determined following intraperitoneal (ip) administration to mice. High-performance liquid chromatography on reversed phase (HPLC-RP) was used also to measure the 60Co irradiation effects on Apis venom. At the end of the study, sheep were killed in a slaughterhouse. Kidneys were histologically analyzed. LD50 was 5.97 mg/kg Apis and 0.07 mg/kg C.d.t. for native compared to 13.44 mg/kg Apis and 0.35 mg/kg C.d.t. for irradiated venoms. HPLC revealed significant differences in chromatographic profiles between native and irradiated Apis venoms. Native venom plus IFA compared with SBA-15 showed significantly higher antibody titers for both venoms. Apis-irradiated venom plus IFA or SBA-15 displayed similar antibody titers but were significantly lower when compared with native venom plus IFA. Weight gain did not differ significantly among all groups. 60Co irradiation decreased toxicity and maintained venom immunogenic capacity, while IFA produced higher antibody titers. SBA-15 was able to act as an adjuvant without producing adverse effects. Hyperimmunization did not affect sheep weight gain, which would considerably reduce the cost of antiserum production, as these sheep were still approved for human consumption even after being subjected to hyperimmunization.

Resposta imune e capacidade de neutralização de anticorpos produzidos em ovinos jovens imunizados com veneno de Crotalus durissus terrificus nativo e irradiado com Cobalto 60

The ELISA technique was used to evaluate and compare young ovine humoral immune response during crotalic antiserum production. Animals were clinically evaluated throughout this process, and the neutralizing capacity of antisera raised against natural (NV) and Cobalt-60 irradiated (IrV) venoms of Crotalus durissus terrificus (C.d.t.) was verified by means of in vitro challenges. Three groups of six animals each were used: G1 received NV; G2 was inoculated with IrV; and G3 was used as control. Animals received six immunizations during 84 days at 14-day intervals. ELISA of antibody profile showed significant difference (p<5%) between experimental groups (G1

Avaliação da resposta humoral e da capacidade de neutralização do soro de camundongos Swiss inoculados com venenos nativo e irradiado com cobalto-60 de serpentes Crotalus durissus terrificus, Bothrops jararaca, Bothrops jararacussu e Bothrops moojeni

Pós-graduação em Doenças Tropicais - FMB

Atividade coagulante e da toxidade da giroxina nativa e irradiada com Cobalto-60 isolada do veneno de Crotalus durissus terrificus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Comparação da resposta imune humoral, da capacidade de neutralização dos soros anticrotálicos produzidos em ovinos jovens, avaliação clínica e ponderal entre animais inoculados com veneno de serpente Crotalus durissus terrificus nativo ou irradiado com Cobalto-60

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Conservação de pêssegos 'Biuti' por irradiação

Este trabalho teve como objetivo o emprego da radiação gama associada ou não à refrigeração na conservação pós-colheita dos pêssegos 'Biuti'. O comportamento destes frutos foi estudado através de análises físicas (perda de massa fresca, coloração da casca, índice de doença, conservação pós-colheita e firmeza) e químicas (pH, acidez total titulável (ATT), sólidos solúveis totais (SST) e o índice de maturidade, expresso pela relação SST/ ATT). Os frutos colhidos no estádio de maturação fisiológica receberam os seguintes tratamentos: 0,0kGy; 0,1kGy; 0,2kGy; 0,3kGy; 0,4kGy e 0,5kGy através do irradiador GAMMABEAN 650 que tem como fonte o Cobalto 60. Os pêssegos foram armazenados em condições ambientais (24±2ºC; 72-76% UR) por 1 semana e em condições de refrigeração em estufa B.O.D. (0±1ºC; 70-80% UR) por 2 semanas. Analisando os resultados pode-se concluir que as diferentes doses de radiação aplicadas não promoveram o aumento da vida-útil pós-colheita de pêssegos. Os frutos armazenados sob refrigeração tiveram maior vida-útil pós-colheita em relação aos armazenados em temperatura ambiente.

Hematological changes in sheep inoculated with natural and Cobalt60-irradiated Crotalus durissus terrificus venom (Laurenti, 1768)

Natural (NV) and Cobalto60-irradiated (IrV) Crotalus durissus terrificus venom were used to evaluate serum production capacity of sheep and possible hematological and biochemical effects. Freeze-dried venom aliquots were diluted in acidified saline solution (NaCl 150 mM, pH 3.0) and irradiated by a Cobalt 60 source at a dose of 5.54 x 102 Gy/h and a concentration of 2.000 Gy. Twelve sheep were divided into two groups of six animals. One group received irradiated venom (IrV) and the other natural venom (NV). Three antigen doses (venom) were administered at monthly intervals. Blood samples were collected weekly for analysis of serum neutralization potency and capacity, complete blood count (CBC), total plasma protein, fibrinogen, albumin, and globulin. At the end of the experiment, the animals were challenged with a LD50 for sheep and showed no signs of envenoming. The two groups did not present clinical alterations. Results of the total leukocyte count did not present interaction or time factor effect for both groups, but there was a different action between them, with the NV group presenting more cells than the IrV group. The leukocyte increase to 13,000/ml indicates that slight leukocytosis occurred in the week after the first inoculation in the NV group. There was no statistically significant difference between groups in the absolute count of segmented neutrophils, eosinophils, and lymphocytes but there were statistically significant oscillations in values at the different collecting times. The NV group presented an increase in the absolute neutrophil count after the first inoculation that persisted for 5 weeks. In the IrV group, the increase in neutrophils occurred only in the first week returning to normal in the following weeks. The alterations in the neutrophil count are indicative of systemic inflammatory response related to cytokine release; response was more marked in the NV group, showing its greater toxicity.

Radiação gama na conservação pós-colheita da nectarina (Prunus persica var. Nucipersica) frigoconservada

O trabalho foi desenvolvido no laboratório de Frutas e Hortaliças do Departamento de Gestão e Tecnologia Agroindustrial da Faculdade de Ciências Agronômicas - UNESP - Câmpus de Botucatu, tendo como objetivo principal a caracterização do comportamento da radiação gama, na conservação pós-colheita da nectarina cv. Sunred. Os frutos foram colhidos no início do estádio de maturação, selecionados, limpos, pré-resfriados (4ºC por 12 horas) e submetidos a diferentes doses de radiação gama, constituindo assim os tratamentos: 1 0,0 kGy, 2 - 0,2 kGy, 3 - 0,4 kGy, 4 - 0,6 kGy, 5 - 0,8 kGy, sendo após armazenados em câmara fria com temperaturas de 0ºC e 90-95% de UR, por 28 dias. As análises foram realizadas a cada sete dias, determinando-se o aspecto visual dos frutos, a perda de massa fresca, a firmeza de polpa, a acidez total titulável (ATT), os sólidos solúveis totais (STT) e a razão STT/ATT. Após 28 dias de armazenamento, verificou-se que os frutos submetidos à dose de 0,4 kGy apresentaram o melhor aspecto visual, as menores perdas de massa fresca, e a maior firmeza de polpa, não ocorrendo, entretanto, variações significativas nos teores de acidez total titulável, sólidos solúveis totais e nos valores da razão SST/ATT.

Laboratory evaluation of young ovines inoculated with natural or 60Co-irradiated Crotalus durissus terrificus venom during hyperimmunization process

Laboratory profile of young ovines was studied in order to evaluate and compare their antiserum production from natural and Cobalt-60 irradiated Crotalus durissus terrificus (C.d.t.) venoms. The parameters analyzed included complete blood count, and urea, creatinine, aspartate aminotransferase, total proteins, albumin and globulin serum measurements. Three groups of six animals each were used. Group 1 (G1) received natural C.d.t. venom; Group 2 (G2) received irradiated C.d.t. venom; and Group 3 (G3) was used as control and did not receive venom, only adjuvants, using seven venom inoculations. During the experimental period, animals were fortnightly weighed. According to clinical and weight evaluation, sheep in post-weaning phase showed no changes in their physiological profiles but had excellent weight gain. The parameters analyzed were not statistically different (p<5%) among the groups tested. The hyperimmunization process was successfully accomplished with the production of specific antibodies against Crotalus durissus terrificus venom. Results bring a new possibility of utilizing ovines in the commercial production of anticrotalic serum, which may be used to treat human and animal envenomation. Its production cost may be reduced by subsequent use of hyperimmunized sheep for human consumption.

Conservação de abacate ‘Hass’ e guacamole por irradiação

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Irradiação e qualidade da carne de frango congelada e embalada à vácuo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of gamma irradiation on the nutritional quality of Agaricus bisporus strains cultivated in different composts

The effect of irradiation doses (0, 125, 250 and 500 Gy) on the nutritional quality of A. bisporus mushrooms (strains ABI-07/06, ABI-05/03 and PB-1) cultivated in composts based on oat straw (Avena sativa) and brachiaria (Brachiaria sp.) was evaluated. The experimental design was 4 x 3 x 2 factorial scheme (irradiation doses x strains x composts), with 24 treatments, consisting of two repetitions each, totaling 48 experimental units (samples of mushrooms). The samples were irradiated in Cobalt-60 irradiator, model Gammacell 220 kGy, with dose rate of 0.740 kGy h(-1), according to the treatments proposed. Subsequently, the control (unirradiated) and the other treatments were maintained at 4 +/- 1 degrees C and 90% RH in a climatic chamber for carrying out the chemical analysis of the mushrooms on the 1st and 14th day of storage. It was found that all A. bisporus strains evaluated were food with excellent nutritional value, because they presented high protein and fiber contents and low ethereal extract content; the chemical characterization of the mushrooms was influenced by the compost type in which they were cultivated; gamma irradiation influenced the chemical composition of mushrooms.

New insights into the structural characteristics of irradiated crotamine

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Irradiação e atmosfera modificada passiva na qualidade pós-colheita de goiabas 'Pedro Sato'

The aim of this study was to evaluate the effect of gamma radiation associated with modified atmosphere on postharvest quality of guavas ‘Pedro Sato’. It was used guavas from the region of Vista Alegre do Alto/São Paulo/Brazil. After harvest, the fruits were immediately transported to the Fruit and Vegetables Laboratory from the Agroindustrial Management and Technology Department, Agronomic Sciences College - UNESP - Botucatu / SP, where they were kept at 10 ° C and 90-95% RH in cold storage, for 28 days. It was used the randomized design, with factorial scheme 5 x 5, three repetitions. The first factor consisted of the following effects: control 1 (without package or irradiation), control 2 (polystyrene package/PS + package low density polyethylene/LDPE and without irradiation), treatment 1 (PS + LDPE and 0.2 kGy ), treatment 2 (PS + LDPE and 0.6 kGy) and treatment 3 (PS + LDPE and 1.0 kGy). The second factor consisted of the evaluation periods: 0, 7, 14, 21 and 28 days. The analyses were: firmness, soluble solids (SS), titratable acidity (TA), maturity index, pH, respiration rate. Concluded that high doses of irradiation promoted a negative effect on physical-chemical characteristics of guava ‘Pedro Sato’, verifying that only the lowest dose associated with modified atmosphere provided fruits with higher quality and acceptability, due to higher maturation rate and soluble solids obtained. Regarding the days of analysis, there were no positive effect of the treatments during storage, where only the early days promoted better values for the variables studied.

Gamma irradiation can control the number of psychotrophic bacteria in Agaricus bisporus during storage

We evaluated the effect of gamma irradiation doses (0, 125, 250, and 500 Gy) in control of psychrotrophic bacteria in different strains of Agaricus bisporus (ABI-07/06, ABI-05/03, and PB-1) during storage, cultivated in composts based on oat straw (Avena sativa) and Brachiaria spp. The experimental design was completely randomized in a factorial scheme 4  2  3 (irradiation doses  composts  strains), with 24 treatments, each consisting of 2 replicates, totaling 48 experimental units (samples of mushrooms). The mushrooms collected from all culture conditions were packaged in plastic polypropylene with 200 g each and subjected to Cobalt-60 irradiator, type Gammacell 220, and dose rate 0.740 kGy h–1 , according to the treatments. Subsequently, the control (nonirradiated) and other treatments were maintained at 4 ± 1°C and 90% relative humidity (RH) in a climatic chamber to perform the microbiological analysis of mushrooms on the 1st and 14th day of storage. According to the results, it was found that the highest mean colony psychotrophic count, after 14 days of storage, was observed in strain ABI-07/06 1.30 × 108 g -1 most probable number (MPN) in nonirradiated mushrooms, coming from Brachiaria grass-based compost, and this same strain under the same storage conditions, coming from the same type of compost that underwent a dose of 500 Gy, obtained a significant reduction in mean colonies of psychrotrophic bacteria (2.25 × 104 g –1 MPN). Thus, the irradiation doses tested favored reducing the number of colonies of psychrotrophic bacteria, regardless of the type of compound and strain of A. bisporus.