Structure, processing and midgut secretion of putative peritrophic membrane ancillary protein (PMAP) from Tenebrio molitor larvae

A cDNA coding for a Tenebrio molitor midgut protein named peritrophic membrane ancillary protein (PMAP) was cloned and sequenced. The complete cDNA codes for a protein of 595 amino acids with six insect-allergen-related-repeats that may be grouped in A (predicted globular)- and B (predicted nonglobular)-types forming an ABABAB structure. The PMAP-cDNA was expressed in Pichia pastoris and the recombinant protein (64 kDa) was purified to homogeneity and used to raise antibodies in rabbits. The specific antibody detected PMAP peptides (22 kDa) in the anterior and middle midgut tissue, luminal contents, peritrophic membrane and feces. These peptides derive from PMAP, as supported by mass spectrometry, and resemble those formed by the in vitro action of trypsin on recombinant PMAP. Both in vitro and in vivo PMAP processing seem to occur by attack of trypsin to susceptible bonds in the coils predicted to link AB pairs, thus releasing the putative functional AB structures. The AB-domain structure of PMAP is found in homologous proteins from several insect orders, except lepidopterans that have the apparently derived protein known as nitrile-specifier protein. Immunocytolocalization shows that PMAP is secreted by exocytosis and becomes entrapped in the glycocalyx, before being released into midgut contents. Circumstantial evidence suggests that PMAP-like proteins have a role in peritrophic membrane type 2 formation. (C) 2007 Elsevier Ltd. All rights reserved.

Microwave-assisted digestion procedures for biological samples with diluted nitric acid: Identification of reaction products

Microwave-assisted sample preparation using diluted nitric acid solutions is an alternative procedure for digesting organic samples. The efficiency of this procedure depends on the chemical properties of the samples and in this work it was evaluated by the determination of crude protein amount. fat and original carbon. Soybeans grains, bovine blood. bovine muscle and bovine viscera were digested in a cavity-microwave oven using oxidant mixtures in different acid concentrations. The digestion efficiency was evaluated based on the determination of residual carbon content and element recoveries using inductively coupled plasma optical emission spectrometry (ICP OES). In order to determine the main residual organic compounds, the digests were characterized by nuclear magnetic resonance (1 H NMR). Subsequently, studies concerning separation of nitrobenzoic acid isomers were performed by ion pair reversed phase liquid chromatography using a C18 stationary phase, water:acetonitrile:methanol (75:20:5, v/v/v) +0.05% (v/v) TFA as mobile phase and ultraviolet detection at 254 nm. Sample preparation based on diluted acids proved to be feasible and a recommendable alternative for organic sample digestion, reducing both the reagent volumes and the variability of the residues as a result of the process of decomposition. It was shown that biological matt-ices containing amino acids, proteins and lipids in their composition produced nitrobenzoic acid isomers and other organic compounds after cleavage of chemical bonds. (C) 2009 Elsevier B.V. All rights reserved.

Anaerobic bio-digestion of concentrate obtained in the process of ultra filtration of effluents from tilapia processing unit

The objective of the present study was to evaluate the efficiency of the process of biodigestion of the protein concentrate resulting from the ultrafiltration of the effluent from a slaughterhouse freezer of Nile tilapia. Bench digesters were used with excrements and water (control) in comparison with a mixture of cattle manure and effluent from the stages of filleting and bleeding of tilapias. The effluent obtained in the continuous process (bleeding + filleting) was the one with highest accumulated population from the 37th day, as well as greatest daily production. Gases composition did not differ between the protein concentrates, but the gas obtained with the use of the effluent from the filleting stage presented highest methane gas average (78.05%) in comparison with those obtained in the bleeding stage (69.95%) and in the continuous process (70.02%) or by the control method (68.59%).

Digestion of feed fractions and intake of heifers fed hydrolyzed sugarcane stored for different periods

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Enolase from Paracoccidioides brasiliensis: isolation and identification as a fibronectin-binding protein

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Functional morphology of adult female Culex quinquefasciatus midgut during blood digestion

The adult female Culex quinquefasciatus midgut comprises a narrow anterior and a dilated posterior region, with epithelia composed of a monolayer of adjacent epithelial cells joined at the apical portion by septate junctions. Densely packed apical microvilli and an intricate basal labyrinth characterise each cell pole. Our morphological studies suggest that, during blood digestion, the anterior midgut region also participates in an initial absorptive stage which is probably related to the intake of water, salts and other small molecules. This activity peaked by 6 h after bloodmeal feeding (ABF) and ended approximately 18 h ABF, when the peritrophic membrane was already formed. After this time, absorption only occurred in the posterior region, with morphologic and biochemical evidence of high synthetic activity related to the secretion of proteases. Chymotrypsin, elastase, aminopeptidase, and trypsin reached their maximum activity at around 36 h ABF. Digestion products were apparently absorbed and transported to the basal labyrinth, from where they should be released to the hemolymph. At 72 h ABF, proteolysis had already ended and protein levels had returned to those observed before blood meal. The epithelium of the posterior region, however, did not return to its initial morphology, appearing quite disorganised. Additionally, from 48 h ABF onwards some epithelial cells showed morphological signals of apoptosis. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Electrophoretic analysis of the protein in the midgut of stingless bees (Apidae; Meliponinae) with a comparison of necrophagous and feeding pollen workers

SDS electrophoresis of midgut proteins from two Trigonini species with different feeding habits shows many similarities and the absence of unique protein in the necrophagous stingless bee Trigona hypogea, suggesting that biochemical adaptation to necrophagy, in this bee, occurs more in the qualitative than in quantitative level. Workers of different ages display some variations in the midgut protein pattern, suggesting that it is important to know the age in studies regarding to bee digestion.

Ruminal parameters analyzed in remaining digestion residue of roughages in the in vitro/gas system

O desempenho animal é a medida mais direta de se avaliar a qualidade dos alimentos. Entretanto, dados de desempenho são insuficientes para se detectar as possíveis interações que possam ocorrer no ambiente ruminal. O objetivo do presente trabalho foi avaliar os possíveis efeitos associativos nas concentrações de ácidos graxos voláteis (AGVs), nitrogênio amoniacal (N-NH3) e pH da fração líquida remanescente da digestão da matéria seca (MS) de volumosos exclusivos (cana-de-açúcar= CN; capim-elefante com 60 dias= CP60 e 180 dias= CP180 de crescimento; e silagem de milho= SIL) e suas combinações (cana-de-açúcar+silagem de milho= CNSIL; cana-de-açúcar+capim-elefante-60d= CNCP60; cana-de-açúcar+capim-elefante-180d= CNCP180; silagem de milho+capim-elefante-60d= SILCP60; silagem de milho+capim-elefante-180d= SILCP180) na proporção de 50% na MS, que levam a resultados de desempenhos positivos ou negativos de bovinos. As concentrações de AGVs, N-NH3 e pH dos tratamentos foram: CN= 56,9 mmol L-1, 50,1 mg dL-1, 5,7; CNSIL= 61,4 mmol L-1, 50,7 mg dL-1, 5,8; CNCP60= 54,7 mmol L-1, 47,6 mg dL-1, 5,8; CNCP180= 45,4 mmol L-1, 49,4 mg dL-1, 6,0; SIL= 57,2 mmol L-1, 54,0 mg dL-1, 5,8; SILCP60= 57,1 mmol L-1, 53,1 mg dL-1, 5,9; SILCP180= 55,9 mmol L-1, 52,3 mg dL-1, 6,0; CP60= 58,1 mmol L-1, 49,4 mg dL-1, 5,9; CP180= 44,0 mmol L-1, 46,4 mg dL-1, 6,1. Os carboidratos não estruturais e amido, aliados à fibra e proteína, contribuíram para que ocorresse o efeito associativo positivo na mistura 50:50 cana/silagem. Isso pode ter propiciado os melhores resultados de desempenho em bovinos devido ao elevado padrão fermentativo.

Ultrastructural evidences of the occurrence of digestion and absorption of organic solutes in bees ileum (Melipona quadrifasciata anthidioides - Hymenoptera, Apidae)

The ultrastructural aspects of the ileum of Melipona quadrifasciata anthidioides, a stingless bee whose workers obtain their protein requirements from pollen, present morphological evidences of digestion and absorption. These evidences are represented by special relationships between food particles and the bacterial nora present and between their assemblage and the ileum wall, mainly in the ileum three anterior fourths. The apical features of the epithelium also are indicative of organic material absorption.

Effect of age on seed digestion in parrots (Amazona aestiva)

The objective of this study was to compare the capacity of adult (more than 3 yr old) and young (less than 1 yr old) true parrots to digest seeds that are normally included in their diet in captivity, particularly soybean, sunflower, and corn. All the seeds were offered for 5 d with an interval of 15 d between different diets. The seeds of soybean and corn were boiled for 15 min and soaked in water at ambient temperature for 12 h before being fed to the birds. There were no differences in the digestibilities of crude protein and fats (ether extract) among animals, but the digestibilities of dry matter and crude fiber by the adult animals were higher than those of the young ones. The digestibility of carbohydrate (nitrogen-free extract) by adult birds was higher only for sunflower seeds. It is concluded that the capacity of parrots to digest fiber may change according to the age of the animal. Since the digestion of fiber depends on the action of microorganisms, these results suggest that the colonization of the gastrointestinal tract is delayed or very slow in young parrots.

Nutrient digestibility of degermed, dehulled corn, citrus pulp, and soy protein concentrate by barrows

This study was carried out to determine apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of DM, CP, GE, and their respective digestible content of degermed dehulled corn (Zea mays), citrus pulp, and soy (Glycine max) protein concentrate by pigs using the difference method. Thirty-two barrows (28.1 +/- 1.6 kg of BW) were fed a corn-soybean meal basal diet or 1 of 3 diets formulated by replacing 30% of the basal diet with 30% of 1 of the test feedstuffs for 11 d. Chromic oxide (0.3%) was included in the diets. Feces were collected from days 7 to 11 by grab sampling and ileal digesta were collected after pigs were slaughtered on day 12. The AID of DM and AID and ATTD of GE of degermed corn (77.4, 88.7, and 77.7%) were greater (P < 0.05) than those observed in citrus pulp (50.3, 86.5, and 55.8%) and in soy protein concentrate (63.5, 85.1, and 59.4%), which did not differ (P > 0.05). The ATTD of CP, total digestible CP, and total DE of soy protein concentrate (87.5%, 500 g/kg, and 3739 kcal/kg) were higher (P < 0.05) than the values in degermed corn (81.7%, 57.5 g/kg, and 3330 kcal/kg), which were greater (P < 0.05) than those in citrus pulp (60.5%, 39.5 g/kg, and 3223 kcal/kg). Total and ileal digestible DM, AID of CP, and ileal DE of degermed corn (782 g/kg, 673 g/kg, 70.7%, and 2913 kcal/kg) and soy protein concentrate (778 g/kg, 570 g/kg, 78.7%, and 2878 kcal/kg) were similar (P > 0.05) and greater (P < 0.05) than those in citrus pulp (737 g/kg, 436 g/kg, 50.6%, and 2081 kcal/kg). Ileal digestible CP of degermed corn (49.8 g/kg) and citrus pulp (33.0 g/kg) did not differ (P > 0.05) but were smaller (P < 0.05) than the value found in soy protein concentrate (434 g/kg). The DM and energy from degermed corn are more efficiently digested by the pig than those from soy protein concentrate and citrus pulp. Soy protein concentrate was the best protein source evaluated in this study.

Restriction endonuclease banding and digestion resistant heterochromatin in triatomines, genus Panstrongylus

The present work realized a comparative study in meiosis of two triatomines, Panstrongylus herreri and P. megistus, by cytogenetic techniques involving the restriction endonucleases Hae III and Alu I and C-banding. The system of sex chromosomes in Panstrongylus is of the X1X2Y type, and experiments corroborated the common origin hypothesis of the X chromosomes by fragmentation of single X. In both species the restriction endonucleases (RE) presented banding patterns in part similar to C-banding. However, in some early meiotic phases it was possible to verify differentiation of the heterochromatic pattern. This work suggests that other elements besides presence of recognition sites, such as chromatin packing degree and DNA-protein interaction, act in RE results, since digestion patterns occur in early spermatogenesis. However, metaphase chromosomes were practically inaccessible to the endonucleases.

Acid and fermented silage characterization and determination of apparent digestibility coefficient of crude protein for pacu Piaractus mesopotamicus

Three types of raw materials including commercial waste from saltwater (SW), freshwater fish (FW) and tilapia fillet residue (FR) were used to produce fish silage by either acid digestion (2% formic acid and 2% sulfuric acid) or anaerobic fermentation (5% of Lactobacillus plantarum and 15% sugar cane molasses). Six test diets were used in digestibility trials prepared with 70% reference diet and 30% of each experimental silage. These diets were fed to juvenile pacu Piaractus mesopotamicus (146 g average weight) in triplicate. Fish were kept in 500-L tanks and feces collected by manual extrusion. It was observed for both processes that SW waste always had the highest moisture content and lowest fat and ash. Highest crude protein levels were found in silages from commercial fish waste (SW and FW) made from whole fish unfit for human consumption. However, apparent digestibility coefficients did not vary among diets (P > 0.05). Although values did not differ statistically, fermented silage consistently displayed higher digestibility coefficients compared to acid silage. The silages exhibited relatively high protein digestibility (72.5-80.0%), thus suggesting the feasibility of using fish industry by-products in aquaculture feeds.

Protein content of leaf-cutting ant queens before the nuptial flight and during the post-claustral phase

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Different true-protein sources do not modify the metabolism of crossbred Bos taurus x Bos indicus growing heifers

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)