Degradabilidade ruminal e digestibilidade intestinal de alimentos por intermédio da técnica in situ associada à do saco de náilon móvel

Objetivou-se no presente trabalho verificar a degradação ruminal e a digestibilidade intestinal e total da matéria seca (MS) e da proteína bruta (PB) do farelo de soja, do grão de milho, do melaço em pó, da farinha de peixe, da farinha de penas e do feno de alfafa, por intermédio da técnica de degradabilidade ruminal in situ associada à técnica do saco de náilon móvel. As amostras dos alimentos foram moídas e colocadas em duplicata em sacos de náilon de 10x5 cm (48 micras) nas quantidades de 15 e 5 mg de MS/cm² para os alimentos concentrados e feno de alfafa, respectivamente. Os sacos de náilon permaneceram incubados no rúmen de bois holandeses por 0; 2; 6; 8; 24 e 48 h; e 0; 8; 12; 24; 48; 72 e 96 horas, respectivamente, sendo depois retirados e sua duplicata inserida no duodeno através de uma cânula. Posteriormente, os sacos foram coletados junto com as fezes. Os valores de degradabilidade efetiva da PB para uma velocidade de passagem de 5%/hora, para o melaço em pó, grão de milho, farelo de soja, farinha de peixe, farinha de penas e feno de alfafa, foram de 100,00; 62,50; 57,90; 39,30; 34,20 e 60,90%, respectivamente; a digestibilidade intestinal de 100,00; 96,05; 99,79; 98,19; 96,07 e 94,64%, respectivamente; e a digestibilidade total de 100,00; 97,86; 99,87; 98,88; 97,35 e 98,09%, respectivamente. Verificou-se que as proteínas do melaço foram totalmente solúveis no rúmen, sendo as do milho, feno e farelo de soja bastante degradadas, além de possuírem um aproveitamento quase total no intestino. As proteínas das farinhas de peixe e de penas apresentaram baixa solubilidade ruminal e alta digestibilidade intestinal, sendo a farinha de peixe levemente mais digerida no intestino do que a farinha de penas.

Evaluation of a novel kit (TF-test) for the diagnosis of intestinal parasitic infections

Intestinal parasitic infections are currently a source of concern for Public Health agencies in developing and developed countries. Since three ovum-and-parasite stool examinations have been demonstrated to provide sensitive results, we designed a practical and economical kit (TF-Test) that is now commercially available (Immunoassay Com. Ind. Ltda., S (a) over tildeo Paulo, Brazil). This kit allows the separate collection of three fecal specimens into a preservative solution. The specimens are then pooled, double-filtered, and concentrated by a single rapid centrifugation process. The TF-Test was evaluated in four different laboratories in a study using 1,102 outpatients and individuals living in an endemic area for enteroparasitosis. The overall sensitivity found using the TF-Test (86.2-97.8%) was significantly higher (P<0.01) than the sensitivity of conventional techniques such as the Coprotest (NIL Comercio Exterior Ltda, São Paulo, Brazil) and the combination of Lutz/Hoffman, Faust, and Rugai techniques (De Carli, Diagnostico Laboratorial das Parasitoses Humanas. Metodos e Tecnicas, 1994), which ranged from 48.3% to 75.9%. When the above combined three specimen technique was repeated with three specimens collected on different days, its sensitivity became similar (P > 0.01) to that of the TF-Test. The kappa index values of agreement for the TF-Test were consistent (P < 0.01), being higher and ranking in a better position than conventional techniques. The high sensitivity, cost/benefit ratio, and practical aspects demonstrate that the TF-Test is suitable for individual diagnosis, epidemiological inquiries, or evaluation of chemotherapy in treated communities. (C) 2004 Wiley-Liss, Inc.

Molecular and biochemical characterization of the Neurospora crassa glycogen synthase encoded by the gsn cDNA

Glycogen synthases catalyze the transfer of a glucosyl moiety from a nucleotide phosphosugar to a nascent glycogen chain via an alpha1-->4 linkage. Although many genes coding for glycogen synthases have been described, the enzymes from rabbit and yeast are the best characterized. The fungus Neurospora crassa accumulates glycogen during exponential growth, and mobilizes it at the onset of stationary phase, or when placed at high temperature or starved for carbon. Through a PCR methodology, the gsn cDNA coding for the N. crassa glycogen synthase was isolated, and the amino acid sequence of the protein was deduced. The product of the cDNA seems to be the only glycogen synthase present in N. crassa. Characterization of the gsn cDNA revealed that it codes for a 706-amino acids protein, which is very similar to mammalian and yeast glycogen synthases. Gene expression increased during exponential growth, reaching its maximal level at the end of the exponential growth phase, which is consistent with the pattern of glycogen synthase activity and glycogen level. Expression of the gsn is highly regulated at the transcriptional level. Under culture conditions that induce heat shock, conidiation, and carbon starvation, expression of the gsn gene was decreased, and glycogen synthase activity and glycogen content behaved similarly.

cDNA cloning and 1.75 angstrom crystal structure determination of PPL2, an endochitinase and N-acetylglucosamine-binding hemagglutinin from Parkia platycephala seeds

Parkia platycephala lectin 2 was purified from Parkia platycephala (Leguminosae, Mimosoideae) seeds by affinity chromatography and RP-HPLC. Equilibrium sedimentation and MS showed that Parkia platycephala lectin 2 is a nonglycosylated monomeric protein of molecular mass 29 407 +/- 15 Da, which contains six cysteine residues engaged in the formation of three intramolecular disulfide bonds. Parkia platycephala lectin 2 agglutinated rabbit erythrocytes, and this activity was specifically inhibited by N-acetylglucosamine. In addition, Parkia platycephala lectin 2 hydrolyzed beta(1-4) glycosidic bonds linking 2-acetoamido-2-deoxy-beta-D-glucopyranose units in chitin. The full-lengthamino acid sequence of Parkia platycephala lectin 2, determined by N-terminal sequencing and cDNA cloning, and its three-dimensional structure, established by X-ray crystallography at 1.75 angstrom resolution, showed that Parkia platycephala lectin 2 is homologous to endochitinases of the glycosyl hydrolase family 18, which share the (beta alpha)(8) barrel topology harboring the catalytic residues Asp125, Glu127, and Tyr182.

Aneuploidies, deletion, and overexpression of TP53 gene in intestinal metaplasia of patients without gastric cancer

Gastric carcinogenesis is attributable to interacting environmental and genetic factors, through a sequence of events including intestinal metaplasia. Using a fluorescence in situ hybridization technique, we investigated the occurrence of ancuploidies of chromosomes 3, 7, 8, 9, and 17, TP53 gene deletion, and expression of p53 in 21 intestinal metaplasia (IM) samples from cancer-free patients and in 20 gastric adenocarcinoma samples. Aneuploidies were found in 71% (15/21) of the IM samples. Trisomy of chromosomes 7 and 9 occurred mainly in complete-type IM; in the incomplete type, trisomy of chromosomes 7 and 8 were more commonly found. The TP53 gene deletion was observed in 60% (3/5) of the IM cases, and immunohistochemistry revealed p53 overexpression in 12% (2/17) of the analyzed IM cases. All gastric adenocarcinoma cases presented higher frequencies of trisomy or tetrasomy of chromosomes 3, 7, 8, 9, and 17. The TP53 deletion was found in all three of the gastric adenocarcinoma analyzed for it, and immunohistochemistry detected overexpression of protein p53 in 80% (12/15) of the analyzed cases. Our study revealed for the first time the presence of aneuploidies of chromosomes 7, 8, 9, and 17 and of TP53 gene deletion and overexpression in IM samples from cancer-free patients. These results suggest that IM and gastric adenocarcinoma may share the same genetic alterations. (C) 2004 Elsevier B.V. All rights reserved.

Intestinal starch disappearance increased in steers abomasally infused with starch and protein

Steers (379 +/- 10 kg) with ruminal, duodenal, and ileal cannulas were used in a 5 x 5 Latin square digestion trial to quantify and evaluate the relationship between intestinal protein supply and intestinal starch disappearance. Treatments were infusions of 0, 50, 100, 150, or 200 g/d of casein along with 1,042 g/d of raw cornstarch. Abomasal infusions were accomplished by passing tubing and a pliable retaining washer through the reticular-omasal orifice into the abomasum. Steers were fed a 93% corn silage, 7% supplement diet that contained 12% crude protein at 1.65% body weight in 12 equal portions/d. Periods lasted 17 d (12 d for adaptation, 2 d of collections, and 3 d of rest). The quantity and percentage of organic matter and protein disappearance from the small intestine increased linearly (P < 0.03) with infused casein. Greater quantities of starch disappeared with increased casein infusion (P < 0.01). The infusion of 200 g/d of casein increased small intestinal starch disappearance by 226 g/d over the control. Casein infusion did not affect the quantity or percent of organic matter, starch, or protein disappearance in the large intestine. Treatments did not change ruminal ammonia N, ruminal pH, or plasma glucose concentrations. Starch disappearance from the small intestine was increased with greater protein flow to the duodenum of steers.

Effects of leucine supplemented diet on intestinal absorption in tumor bearing pregnant rats

Background: It is known that amino acid oxidation is increased in tumor-bearing rat muscles and that leucine is an important ketogenic amino acid that provides energy to the skeletal muscle.Methods: To evaluate the effects of a leucine supplemented diet on the intestinal absorption alterations produced by Walker 256, growing pregnant rats were distributed into six groups. Three pregnant groups received a normal protein diet (18% protein):pregnant (N), tumor-bearing (WN), pair-fed rats (Np). Three other pregnant groups were fed a diet supplemented with 3% leucine (15% protein plus 3% leucine):leucine (L), tumor-bearing (WL) and pair-fed with leucine (Lp). Non pregnant rats (C), which received a normal protein diet, were used as a control group. After 20 days, the animals were submitted to intestinal perfusion to measure leucine, methionine and glucose absorption.Results: Tumor-bearing pregnant rats showed impairment in food intake, body weight gain and muscle protein content, which were less accentuated in WL than in WN rats. These metabolic changes led to reduction in both fetal and tumor development. Leucine absorption slightly increased in WN group. In spite of having a significant decrease in leucine and methionine absorption compared to L, the WL group has shown a higher absorption rate of methionine than WN group, probably due to the ingestion of the leucine supplemented diet inducing this amino acid uptake. Glucose absorption was reduced in both tumor-bearing groups.Conclusions: Leucine supplementation during pregnancy in tumor-bearing rats promoted high leucine absorption, increasing the availability of the amino acid for neoplasic cells and, mainly, for fetus and host utilization. This may have contributed to the better preservation of body weight gain, food intake and muscle protein observed in the supplemented rats in relation to the non-supplemented ones.

EFFECTS OF PROTEIN-DEFICIENCY AND NATURAL INTESTINAL INFECTION WITH GIARDIA-LAMBLIA ON JEJUNAL INTRAEPITHELIAL LYMPHOCYTES IN RATS OF DIFFERENT AGES

1. The interaction between experimental protein deprivation and natural intestinal infection by Giardia lamblia was studied in terms of its effects on the intraepithelial lymphocyte (IEL) population and morphology of the jejunal mucosa of rats of different ages.2. Young, adult and old male Wistar rats received a protein-deficient diet (2% casein) or a control diet (20% casein) for 42 days. Mucosal height and the number of lymphocytes located among 500 consecutive epithelial cells (EC) along the villi or crossing the basement membrane were determined in PAS-stained jejunal fragments.3. The number of IEL increased progressively with animal age, from 14 to 25 per 100 epithelial cells, with significant differences between age ranges. However, the number of IEL did not differ between control and protein-deficient rats in any of the age groups. The proportion of lymphocytes crossing the basement membrane was approximately two-fold greater in young (2.8/100 EC) and adult (5.8/100 EC) protein-deficient animals than in their respective controls (1.6 and 2.8/100 EC). The intensity of parasite colonization was moderate, from 3 to 5/100 EC and did not differ between groups. The pattern of morphologic changes of jejunal mucosa in protozoal infection did not differ between control and protein-deficient animals in any of the three age groups.4. We conclude that intestinal infection with Giardia lamblia probably stimulated the local immune response, masking the reduction of the IEL population induced by protein deficiency. The increase in lymphocyte numbers with age may be related to prolonged antigenic stimulation promoted by infection.

INTERFERENCE BETWEEN SALMONELLA SEROTYPES IN INTESTINAL COLONIZATION OF CHICKENS - CORRELATION WITH IN-VITRO BEHAVIOR

The effect of colonisation of the alimentary tract of newly hatched chicks by different Salmonella serotypes on the establishment in the gut by other Salmonella strains inoculated afterwards was assessed. Although profound inhibition of colonisation had been found previously to be genus-specific, considerable variation was found within the Salmonella genus. Some strains were found to be much more inhibitory than others and some were more easily inhibited than were others. There was not an absolute relationship between inhibitory activity and colonisation ability. No relationship was seen between inhibition and serotype or phage types within serotypes. There was no correlation between in vivo inhibition and the extent of inhibition that occurred in early stationary phase cultures in rich, undefined broth cultures.

Intestinal antiinflammatory activity of a lyophilized infusion of Turnera ulmifolia in TNBS rat colitis

Turnera ulmifolia is a plant popularly known in Brazil and South America as chanana. Some species of Turnera are widely used in folk medicine for different types of inflammatory diseases. In this study, the preventive intestinal antiinflammatory activity of a lyophilized infusion obtained from the aerial parts of T. ulmifolia was tested in the trinitrobenzenesulphonic acid (TNBS) model of rat colitis. The results obtained revealed that pretreatment to colitic rats with the extract, at 250 and 500 mg/kg, significantly attenuated the colonic damage induced by TNBS. This beneficial effect was associated with an improvement in the colonic oxidative status, since the infusion prevented the glutathione depletion that occurred as a consequence of the colonic inflammation. on the other hand, this antioxidant activity was confirmed in in vitro studies. In conclusion, the preventive effect exerted by the lyophilized infusion of T. ulmifolia in the TNBS model of rat colitis is probably related to its antioxidant properties, due to its flavonoids content. (c) 2006 Elsevier B.V. All rights reserved.

SEQUENCE OF A CDNA-ENCODING BOTHROPSTOXIN-I, A MYOTOXIN FROM THE VENOM OF BOTHROPS-JARARACUSSU

With the aim of further understanding the structure/function relationships in the membrane-damaging activity of the Lys(49) phospholipase A(2) (Lys(49)-PLA(2)) sub-family, we used PCR (polymerase chain reaction) on total venom gland cDNAs from Bothrops jararacussu with degenerate oligodeoxyribonucleotides encoding the N- and C-termini of myotoxin II, a Lys(49)-PLA(2) from Bothrops asper. A 350-bp cDNA coding for bothropstoxin I (BtxtxI) was amplified. Sequencing of the amplified fragment shows that BtxtxI has a Lys(49), and comparison with the known structure of myotoxin II showed that the amino acids involved in the formation of a novel dimeric structure in this protein were also conserved.

Posthatching water and feed deprivation affect the gastrointestinal tract and intestinal mucosa development of broiler chicks

We studied the effect of feed and water deprivation on gastrointestinal tract and intestinal mucosa development of chicks at 24, 48, and 72 h posthatching. The treatments were water and feed ad libitum, water ad libitum and no feed, no water but feed ad libitum, and no water and no feed. The relative weight of the yolk sac was not influenced by the treatments. However, at 48 and 72 h posthatching, the relative weight of the liver increased, and the gizzard + proventriculus weight decreased in birds receiving feed ad libitum. An increase in jejunum and ileum relative weights and lengths was observed when the birds were supplied with feed and water. The lack of water produced the same effect as the lack of feed, both causing a higher number of villi per area with reduction in villus size, when compared with feed and water ad libitum treatments. The results of this study revealed that feed and water are able to affect intestinal villus development after hatching, indicating that both feed and water must be supplied to the chicks immediately after hatching.

Anti-inflammatory intestinal activity of Abarema cochliacarpos (Gomes) Barneby & Grimes in TNBS colitis model

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Morphometric study of the small intestinal mucosa in young, adult, and old rats submitted to protein deficiency and rehabilitation

Linear and stereological morphometric methods were applied to the jejunal and ileal mucosa of young, adult, and old male Wistar rats submitted to protein deficiency and rehabilitation. The animals were fed ad libitum a 2% casein diet during 42 days and then received a 20% casein diet for 30 days. Food intake, body weights, and plasma protein concentrations were recorded. In the young protein deficient rats values of mucosal height, surface area, and volume of the lamina propria were significantly lower than those of their age controls in both jejunum and ileum. In adults the differences were less marked and in the old rats all parameters were found to be unaltered by the protein deficient diet. The surface-to-volume ratio showed no significant differences between control and protein deficient in all three age groups, meaning that villus pattern did not change with protein deficiency. On rehabilitation, a striking difference between jejunum and ileum was observed in the young rats; all parameters returned to control levels in the jejunum, while they remained lower than those of their controls in the ileum.