不同盐度对弓斑东方鲀消化酶活性的影响

本研究探讨了不同水体盐度对弓斑东方鲀α-淀粉酶、胃蛋白酶和胰蛋白酶活性的影响,同时得出最适于该鱼种消化吸收养分的盐度值,以发挥其生长潜力.试验用360尾鱼,分为4个处理(淡水对照,盐度8‰、18‰和35‰),每处理3个重复,每个重复30尾,通过15 d饲养试验,采样时间分别为:第1天、第4天、第7天和第15天.结果显示:随着盐度的增加,α-淀粉酶、胃蛋白酶和胰蛋白酶的活性有显著变化(P<0.05).在试验后期,α-淀粉酶活性在盐度18‰时显著高于盐度8‰和35‰的情况(P<0.05);胃蛋白酶活性则在8‰和18‰2种盐度下有显著差异(P<0.05);胰蛋白酶活性在8‰时最大,显著高于35‰和18‰处理(P<0.05).试验结果表明弓斑东方鲍饲养的水体盐度为8‰～18‰时能更有效的促进其消化酶的活性.     

Effect of predigestion factors on the apparent digestibility of protein for swine determined by the Mobile Nylon Bag Technique

Five Yorkshire x Lacombe barrows (45 kg initial wt) fitted with duodenal cannulas approximately 10 cm from the pyloric sphincter were used to determine the influence of the pH of the predigestion solution (pH 1.0, 1.5, 2.0 or 2.5), pepsin activity (189 vs 377 IU/liter) and duration of predigestion (0, 1.5, 2.5 or 4.0 h) on the apparent digestibility of the protein in soybean meal (SBM), meal and bone meal (MBM) or canola meal (CM) determined with the Mobile Nylon Bag Technique (MNBT). An additional six barrows were used to determine protein digestibility by conventional methods. Protein digestibilities determined using conventional digestibility techniques were 93.1 +/- .7, 79.1 +/- 1.8 and 79.3 +/- 1.4% for the SBM, MBM and CM diets, respectively. Protein digestibilities determined with the MNBT were highest at pH 2.0 for all three protein sources. Increasing pepsin activity from 189 to 377 IU/liter resulted in a slight increase in the digestibility of SBM and CM, but not of MBM. The absence of predigestion (0 h) resulted in a dramatic reduction in protein digestibility; predigestion times of 1.5 or 2.5 h usually resulted in lower protein digestibilities than did a predigestion time of 4.0 h. The closest agreement between results obtained by the MNBT and conventional digestibility studies occurred with a pH of 2.0, a predigestion time of 4.0 h and a pepsin activity of 377 IU/liter.     

Effect of the consumption of Lysiloma latisiliquum on the larval establishment of gastrointestinal nematodes in goats

The consumption of tannin-rich (TR) forages has been associated with negative effects against gastrointestinal nematodes and with an improved host resilience. It has been hypothesized that tannins affect the capacity of infective larvae to establish in the mucosae of the host. In this study, we aimed at testing this hypothesis using Lysiloma latisiliquum, a tropical TR tree. The objectives were: (i) to evaluate the effect of the consumption of L. latisiliquum on the establishment of nematode third-stage larvae (L3) in goats; (ii) to define the role of tannins in these effects in vivo by using an inhibitor (polyethylene glycol, PEG); and (iii) to examine a possible indirect effect of tannins on the inflammatory response in the digestive mucosa. Eighteen Criollo goats composed three experimental groups. The control group received fresh leaves of Brosimum alicastrum, a plant with a low level of tannins. Two groups received L. latisiliquum leaves either with (L.L.+PEG) or without (L.L.) daily addition of 25g PEG. After a 7-day adaptation period, each goat was infected with both Haemonchus contortus and Trichostrongylus colubriformis (3000 L3 per species). The goats were slaughtered 5 days after infection and worm counts and histological analyses were performed. No difference in the voluntary feed intake of foliage was observed between the 3 groups. The consumption of L. latisiliquum significantly reduced the larval establishment of both nematode species compared to the control (P<0.01). For both worm species, the effects were totally alleviated with PEG (L.L.+PEG group), suggesting a major role of tannins in the observed effects. Only minor differences in the mucosal cellular response were observed between the 3 groups. These results confirm that the consumption of TR plants reduces the establishment of nematode larvae in the host and that a direct effect is principally involved.     

胃蛋白酶和胰蛋白酶对大豆凝集素的稳定性研究

本研究以纯化的大豆凝集素(SBA)为研究对象,体外研究了动物体内两大主要蛋白消化酶即胃蛋白酶和胰蛋白酶对其酶解作用。试验通过SDS-PAGE方法分析了SBA经蛋白酶酶解后的降解程度;利用间接竞争ELISA方法测定其在酶解过程中的活性变化。结果表明:SBA可缓慢地被胃蛋白酶降解,SBA浓度越高,完全降解需要的时间越长;随着SBA的降解,SBA的活性也逐渐减小,随着酶解时间的延长活性消失速度也逐渐变慢,并且在本研究选取的酶浓度作用下,SBA浓度越高,活性减小的速度越快,但活性完全消失的时间延长。然而,SBA不能被胰蛋白酶降解,并且酶解过程中SBA的活性也保持完好,说明SBA对胰蛋白酶具有很好的稳定性。因本研究结果将为进一步揭示大豆凝集素在动物体内的消化动力学及其对机体的抗营养作用机理研究奠定理论基础。     

Comparative studies of versatile extracellular proteolytic activities of lactic acid bacteria and their potential for extracellular amino acid productions as feed supplements

Background: Increasing understanding on the functions of amino acids(AA) has led to new commercial applications and expansion of the worldwide markets. However, the current technologies rely heavily on non-food grade microorganism and chemical synthesis for the production of AA. Several studies reported that lactic acid bacteria(LAB) have the capability of producing AA owing to their well-established proteolytic system and amino acid biosynthesis genes. Hence, the objectives of this study were to explore the extracellular proteolytic activity of LAB isolated from various Malaysian fermented foods and their potential to produce AA extracellularly as feed supplements.Results: Al the studied LAB isolates were versatile extracel ular protease producers, whereby extracel ular protease activities were detected from acidic to alkaline pH(pH 5, pH 6.5, pH 8) using qualitative and quantitative proteolytic assays.The highest proteolytic activity at pH 5(15.76 U/mg) and pH 8(19.42 U/mg) was achieved by Lactobacil us plantarum RG14, while Lactobacil us plantarum RS5 exhibited the highest proteolytic activity of 17.22 U/mg at pH 6.5. As for the results of AA production conducted in de Man, Rogosa and Sharpe medium and analysed by high pressure liquid chromatography system, al LAB isolates were capable of producing an array of AA. General y, Pediococcus sp. showed greater ability for AA production as compared to Lactobacil us sp. Moreover, the studied LAB were able to produce a few major feed supplement AA such as methionine, lysine, threonine and tryptophan. P. pentosaceus TL-3 recorded the highest methionine and threonine productivity of 3.72 mg/L/h and 5.58 mg/L/h respectively. However, L. plantarum I-UL4 demonstrated a lysine productivity of 1.24 mg/L/h, while P. acidilactici TP-6 achieved up to 1.73 mg/L/h of tryptophan productivity.Conclusion: Al the 17 studied LAB isolates possessed versatile extracel ular proteolytic system and have vast capability of producing various amino acids including a few major feed supplement AA such as methionine, lysine, threonine and tryptophan. Despite AA production was strain dependent, the studied LAB isolates possessed vast potential and can be exploited further as a bio-agent or an alternative amino acids and bioactive peptide producers.     

Parasitism between Anisakis simplex (Nematoda: Anisakidae) third-stage larvae and the spotted mackerel Scomber australasicus with regard to the application of stock identification

The nematode fauna of 369 spotted mackerel of the species Scomber australasicus, collected off the northeastern Taiwanese coast of the northwestern Pacific, was investigated monthly from April 2004 to March 2005. The following nematode species were recorded: Anisakis simplex complex, Hysterothylacium aduncum, Porrocaecum decipiens and Raphidascaris trichiuri. The seasonal variation in the infection with A. simplex third stage larva (L3) was studied throughout the 12 months. The prevalence of A. simplex L3 recorded for total fish samples was 93.6%, varying between 86.7 and 100%. There was an increase in the abundance of this nematode in spring, with the peak occurring in April. To reveal whether intrinsic factors of the spotted mackerel host contributed to infection with this nematode, fish were grouped according to their body weight, age and gonad development (reported as gonadoosomatic index, GSI), respectively, and infection parameters (i.e., prevalence, abundance and intensity) were analyzed. Results showed that abundance was significantly higher in both larger (>450 g) and older (>3 years old) fish. The gonad development of the host fish was not correlated with the intensity of the larval infection in both female and male fish. Two distinct Anisakis species were identified by PCR-RFLP, namely A. pegreffii and a recombinant genotype of A. pegreffii and A. simplex sensu stricto. These species occurred with frequencies of 97% and 3%, respectively. The usefulness of using parasites as biomarkers for spotted mackerel stock identification around Taiwanese waters was confirmed herein. A second group of 58 spotted mackerel were obtained from the coastal waters off southwestern Taiwan. In addition to the two species, A. pegreffii and the recombinant one, which were found with frequencies of 63% and 9%, respectively, an additional Anisakis species A. typica was identified with a frequency of 28% from these fish. Two spotted mackerel stocks could thus be identified based on their infrastructure of Anisakis species community and their frequency. To the best of our knowledge, this is the first report of stock identification of spotted mackerel using endoparasite biomarkers.     

Proteolytic activity in salivary gland products of sheep bot fly (Oestrus ovis) larvae

This study identified and characterized hydrolytic enzymes in salivary gland products of Oestrus ovis larvae. Third instars were collected from the heads of slaughtered goats. Salivary glands were extracted, their products obtained by centrifugation and the enzymatic profile determined. Optimum pH, temperature of maximum proteolytic activity, thermal stability, and resistance of salivary gland products were determined on collagen and subclasses of proteases were identified using protease inhibitors. Zymograms were used to determine the molecular weight of proteases. Antigenic protein bands were revealed by immunoblotting using sera obtained from experimentally infested goats. Seven positive enzymatic activities were detected in salivary gland products: acid phosphatase, naphthol-AS-BI-phosphohydrolase, esterase (C4), esterase lipase (C8), leucine arylamidase, alpha-glucosidase and N-acetyl-beta-glucosaminidase. Optimum pH for proteolytic activity was 8.0; proteolytic activity increased with temperature (10-50 degrees C) then drastically decreased at 60 degrees C. Proteases in O. ovis salivary gland products belong to the serine subclass. In Zymograms, bands of proteolytic activity were detected in the 20-63 kDa range; the immunoblot showed three antigenic bands, one of them related to a protease band (63 kDa). Serine proteases in O. ovis salivary gland products are most likely involved in larval nutrition and host immuno-modulation.     

Production of eosinophil chemoattractant activity by ovine gastrointestinal nematodes

Eosinophilia is a well documented feature of helminth infections but the precise nature of the interaction between parasite and eosinophil remains an enigma. This paper describes experiments demonstrating that ruminant gastrointestinal trichostrongyles produce potent chemoattractant activity for ovine bone marrow-derived eosinophils in vitro. This activity was initially identified as a constituent of whole worm extracts of third and fourth larval (L3, L4), and adult stages of Teladorsagia circumcincta, and adult Haemonchus contortus. Similar activity was detected in excretory/secretory (E/S) material derived from live T. circumcincta L3. Subsequently, by adapting the assay technique to incorporate live worms directly into the system, it was shown that L3 of both T. circumcincta and H. contortus produced eosinophil chemoattractant activity. In contrast, neither whole worm extracts, or E/S preparations from mixed stages of the free-living nematode Caenorhabditis elegans contained eosinophil chemoattractant activity, and there was no evidence of chemoattractant production by live C. elegans. The results described are challenging to the traditional dogma that eosinophils are host-protective effector cells, and raise the intriguing possibility that ovine nematodes actively encourage recruitment of eosinophils. Local eosinophil-mediated mucosal damage, comparable to that seen in the asthmatic lung, may then provide a permissive local microenvironment for the parasite. Moreover, if they prove important for pathogenicity, nematode chemoattractants could offer future potential as novel therapeutic targets.     

Preliminary characterisation of extracellular serine proteases of Dermatophilus congolensis isolates from cattle,sheep and horses

Dermatophilus congolensis is a filamentous branching actinomycete that causes dermatophilosis, an exudative dermatitis in ruminants. The pathogenesis of this disease is poorly understood and virulence factors of D. congolensis have not been characterised. Culture filtrate (CF) of 14 D. congolensis isolates from cattle, 15 from sheep and four from horses were examined for proteolytic activity using azocasein as a non-specific substrate. The isolates were from a variety of geographical locations. All the isolates examined produced extracellular proteolytic activity. CF from 24 and 48 h cultures and from first and third passages contained proteases. Proteolytic activity was greatest in neutral to alkaline pH (pH 7–10). CF of bovine isolates contained more proteolytic activity than that of ovine isolates. Furthermore, in substrate SDS-PAGE gels containing azocasein the number of proteolytic bands and their molecular weights in CF of bovine, ovine and equine isolates were different, giving distinctive band patterns for isolates from each host species. Three out of four bovine isolates from Antigua gave a fourth band pattern. Bands of equivalent molecular weights to the proteases could not be identified in silver stained SDS-PAGE gels of CF. Serine protease inhibitors had a concentration-dependent inhibitory effect on proteolytic activity in CF and inhibited activity of all proteolytic bands in substrate gels. With the exception of EDTA which had a variable-enhancing effect on activity, inhibitors of other classes of protease had no effect on activity. We conclude that D. congolensis produces a number of extracellular alkaline serine proteases, our results suggest the presence of host-specific variation between isolates and to a lesser extent between isolates from the same host species.     

鸡、鸭消化道pH和消化酶活的比较研究

为阐明鸡、鸭对营养物质利用率差异的原因，比较了8只公鸡和8只公鸭消化道内容物酸碱性及体内主要蛋白质消化酶、碳水化合物水解酶和脂肪水解酶活性的差异。结果表明：（1）鸭消化道前段除口腔pH比鸡的高（P〈0.01），食管膨大部、腺胃和肌胃的pH均比鸡低（P〈0.05或P〈0.01），而后段消化道从空肠开始到直肠，鸭的pH均比鸡高（P〈0.05）。（2）鸡和鸭消化道内主要蛋白酶活性存在差异。鸭肌胃内容物中胃蛋白酶的相对活性和总量都极显著高于鸡的（P〈0.01）；鸭十二指肠、空回肠和盲肠内容物以及胰腺组织中胰蛋白酶的相对活性和总量高于鸡（P〈0.05或P〈0.01）；鸭空回肠内容物中糜蛋白酶的活性显著高于鸡（P〈0.05）。（3）鸭胰腺组织中脂肪酶相对活性极显著高于鸡（P〈0.01），脂肪酶总量显著高于鸡（P〈0.05）；肠道内容物中脂肪酶的活性也大多数高于鸡。（4）鸡和鸭消化道内可溶性碳水化合物水解酶活性差异没有很强的规律性，但鸭空回肠内容物中纤维素酶的相对活性高于鸡（P〈0.01），盲肠内容物中纤维素酶的总量显著高于鸡（P〈0.05）。消化道中消化酶活性差异是鸡鸭对饲料养分消化利用差异的原因之一。     

Survival of dietary antigens in the digestive tract of calves intolerant to soybean products

Preruminant calves were given a series of feeds containing heated soybean flour. Digesta collected from the small intestine were analysed by haemagglutination inhibition assay for soluble soybean antigens. The presence of undenatured glycinin and beta-conglycinin in ileal contents was associated with digestive disturbances. In vitro experiments showed that, under optimal conditions of pH for protease activity, beta-conglycinin but not glycinin, was unaffected by pepsin and both antigens were resistant to rennin and trypsin. The solubility of glycinin and beta-conglycinin in saline extracts remained high over pH ranges of 2 to 3 and 6 to 10 but under conditions of intermediate pH, about 4.5, solubility was minimal. It was concluded that preruminant calves suffer from gastrointestinal hypersensitive responses to certain soybean products because major proteases of the digestive tract fail to denature soluble antigenic constituents of the soybean protein.     

Digestive enzymes along digestive tract of a carnivorous fish Glyptosternum maculatum (Sisoridae,Siluriformes)

This study attempted to determine the presence, distribution and levels of digestive proteases, pepsin, amylase and lipase in the homogenates of digestive tract of a freshwater, cold‐adapted, carnivorous teleost of Tibet, Glyptosternum maculatum (n = 10, mean weight: 129.6 ± 28.9 g, mean length: 195.1 ± 16.6 mm, approximately 6 years old), at different temperatures and pH levels. The descending order of protease activity was as follows: stomach (pepsin, 16.16 ± 0.96 U/mg protein), anterior intestine (3.18 ± 0.25 U/mg protein), posterior intestine (1.76 ± 0.21 U/mg protein) and middle intestine (1.52 ± 0.23 U/mg protein). Amylase activity levels in descending order were as follows: anterior intestine (0.0062 ± 0.0007 U/mg protein), stomach (0.0032 ± 0.0009 U/mg protein), middle intestine (0.0023 ± 0.0005 U/mg protein) and posterior intestine (0.0023 ± 0.0004 U/mg protein). The highest lipase activity was found in the anterior intestine (0.83 ± 0.25 U/mg protein), followed by posterior intestine (0.51 ± 0.19 U/mg protein), middle intestine (0.48 ± 0.09 U/mg protein) and stomach (0.39 ± 0.10 U/mg protein). The optimal temperature and pH level for protease were 50 °C and 9.0–10.0, respectively, along the intestine. The pepsin optima in the stomach were 30 °C and 2.0 respectively. The optimal temperature for amylase was found to be 30 °C along the digestive tract and the optimal pH was 7.0 in the intestine and 6.0 in the stomach. The optimal temperature and pH levels of lipase were 30 °C, pH 6.0 for the stomach and 40–50 °C, pH 8.0 for the intestine, respectively. Hepato‐somatic index was 1.35 ± 0.2% and relative intestine length was 0.90 ± 0.19. The weight ratio of the stomach and intestine to body weight was 1.63 ± 0.43% and 1.57 ± 0.24% respectively.     

The Effects of Peptide-Precipitating Lignosulphonic Acids on the Proteolytic Activity of Pepsin in Vitro and on the Response of Pigs to an Ulcer Inducing Diet

The inhibitory effect of peptide-precipitating lignosulphonic acids on the proteolytic activity of pepsin in vitro has been studied. When an unfractionated lignosulphonic acid preparation was used, total inhibition of the activity of 5 µg crystalline pepsin was obtained by addition of about 15 mg of lignosulphonic acids.The greatest inhibitory effect was seen with the high-molecular weight lignosulphonic acids, but an effect was also seen with middle-and low-molecular weight lignosulphonic acids.A reduction in the inhibitory effect was seen when the experiments were performed in the presence of arginine. The inhibition mechanism is discussed in relation to the peptide-precipitating ability of the lignosulphonic acids.No obvious protection against the appearance of stomach ulcers in four pigs receiving an ulcerogenic diet was seen.     

Proteolytic enzymes from Trichinella spiralis larvae.

Trichinella spiralis larvae infect their hosts by the penetration of small intestine enterocytes. The exact mechanism of penetration is unknown, but the presence of proteolytic enzymes is suspected. In this study, whole worm extracts and excretory-secretory (ES) components were obtained and their proteolytic enzymes examined. Enzymes from worm extracts were capable of hydrolysing azocoll, a general protease substrate in a wide range of pH (2-8), with maximal activity at pH 5. Trichinella spiralis larval enzymes were sensitive to metalloprotease and serine protease inhibitors. Three proteases were identified in worm extracts at molecular weight (MW) 48, 54 and 62 kDa by incorporating a gelatine substrate into a standard or a modified sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) set-up, in which we used low SDS concentration in gel and electrophoresis buffer (0.01%). Intact larvae incubated in a medium containing azocoll showed azocollytic activity. Subsequent analysis of ES products by modified SDS-PAGE in gels containing gelatine demonstrated the presence of three protease of apparent MW 33, 62 and 230 kDa.     

Toxocara canis: proteinases in perivitelline fluid from hatching eggs

Developing larvae of Toxocara canis may secrete several kinds of enzymes within the egg perivitelline fluid (EPF) prior to and during hatching. In particular, proteinases in EPF could play a role in larval emergence within the host gastrointestinal lumen but its presence and nature is unknown. In this work, proteolytic activities in hatching fluid of T. canis were identified and analysed by substrate gel electrophoresis at different pH values and by using type specific protease inhibitors. Three bands of 91, 68 and 38 kDa showed gelatinolytic activity and all proteinase activity from EPF was of the aspartic-type since it was inhibited by pepstatin A. Interestingly, a significantly higher proteolytic activity was observed at acidic pH (< or =5.5). These data suggest that T. canis developmentally secretes and accumulates in EPF aspartic proteinases with a pH-dependent activity that might help the parasite to take advantage of conditions in the host gastrointestinal microenvironment where egg hatching is induced and executed.     

Differences in the digestive tract characteristics of broiler chickens fed on complete pelleted diet or on whole wheat added to pelleted protein concentrate

1. The effects of whole grains of wheat on the digestive tract of broiler chickens was studied. A complete pelleted feed was compared with free choice feeding of whole wheat and a pelleted protein concentrate, given from 7 to 29 d of age. 2. Pepsin activity in proventriculus tissue was lower in whole wheat-fed birds than in complete diet-fed birds. The weight (g/kg body weight) of the gizzard was higher in whole wheat-fed birds and its contents had a lower pH. 3. In the intestine, there were no differences in deoxyribonucleic acid (DNA) concentration, protein/DNA, ribonucleic acid (RNA)/DNA, RNA/protein ratios or alkaline phosphatase activity expressed per tissue weight. The weight (g/kg body weight) of the duodenum was lower in whole wheat-fed birds and its contents had a higher pH. Also the activities of alkaline phosphatase and leucine aminopeptidase in the duodenum, and maltase in the ileum, expressed per unit of bird weight, were lower in whole wheat-fed birds. 4. These results suggest that whole grain feeding increases the chemical (pepsin in proventriculus) and physical (gizzard muscle) functionality of the upper part of the digestive tract but decreases the digestive capacity of the intestine. Higher gizzard functionality may play a positive role in the control of bacterial populations. The lower digestive enzyme activities in the intestine may be detrimental in situations of mucosal deterioration caused by intestinal disease.     

复合营养舔砖对瘤胃酶活性及消化代谢的影响

为了研究复合营养舔砖对瘤胃酶活性及消化代谢的影响,选用4头装有永久瘤胃瘘管的公牛(丹麦红牛公与闽南黄牛母的杂种一代),采用反转试验设计,对瘤胃内pH值、氨态氮(NH3-N)和瘤胃液中纤维素酶、蛋白水解酶的活性进行测定。研究表明:(1)饲喂舔砖后,瘤胃液pH值在短时间内下降,随后又缓慢升高。pH值波动幅度较大,这与复合营养舔砖的蛋白降解率较高有关。(2)舔砖对瘤胃内NH3-N浓度有较大影响,舔砖的粗蛋白含量高达38.7%,产生的NH3-N浓度比对照组高且差异显著。(3)舔砖对瘤胃液蛋白水解酶和纤维素酶的活性没有显著影响,这表明每头饲喂500g/d舔砖对牛的消化代谢功能不会产生不良影响。     

细胞培养基MLM-45及培养条件对不同发育时期柞蚕卵巢细胞生长的影响

选择柞蚕5龄幼虫以及前滞育期、滞育期和解除滞育后发育前期、中期、后期的柞蚕蛹分离卵巢细胞,利用新研制的柞蚕细胞培养基MLM-45,在不同温度、pH值及渗透压条件下进行柞蚕卵巢细胞的体外培养,探究最佳培养条件及卵巢细胞取材的最佳发育时期。用台盼蓝染色活细胞计数方法,测定不同条件下用MLM-45培养基培养的不同发育时期柞蚕卵巢细胞的活力,当在27℃、pH 6.3、渗透压325 mOsm/kg的条件下培养30 d时,上述各发育时期柞蚕卵巢细胞的存活率最高,并且5龄幼虫、前滞育期和解除滞育后发育前期的卵巢细胞出现了增殖,其增殖率分别为54%、64%和2%,滞育期及解除滞育后发育中期和后期的卵巢细胞存活率也分别达到99.6%、96.6%和92.4%。在此最佳培养条件下,各发育时期柞蚕卵巢细胞培养2个月后其形态以圆形和椭圆形为主,尤以5龄幼虫和前滞育期蛹卵巢细胞的培养效果最好,细胞增殖7～8倍,是适于以MLM-45培养基培养的最佳发育时期柞蚕卵巢细胞。     

Chicken egg antibodies for prophylaxis and therapy of infectious intestinal diseases. II. In vitro studies on gastric and enteric digestion of egg yolk antibodies specific against pathogenic Escherichia coli strains

Specific antibodies directed against enterotoxic E. coli strains from the yolk of immunized hens were exposed in vitro to the influence of varying digestive phenomena like reduction of pH and proteolytic digestion. The remaining antibody activity was tested in a specific ELISA system. It could be shown that already within the stomach a considerable loss in antibody activity caused by a lowering of pH and peptic cleavage can be expected. A further loss in antibody activity is due to the proteolytic effect of the pancreas proteases trypsin and chymotrypsin. It was found that antibodies in protein rich solutions like egg or yolk suspensions were more resistant than mere globulin fractions or antibodies isolated by affinity chromatography. Prospects for further in vivo tests are discussed.     

The effect of microbial and animal proteinases on peptide- and protein-lignosulphonic acid complexes in agar gel

It is shown that various microbial and animal proteinases, with the exception of pepsin, have a lytic effect on peptide- and protein-lignosulphonic acid complexes in agar gel, and that this effect is characteristically inhibited by specific antiproteinases and naturally occurring serum inhibitors. The activity of the enzymes is studied at various concentrations of peptides and proteins, and at various pH values in the agar. The lysis phenomenon is seen in relation to the precipitation theory of peptide- and protein-lignosulphonic acid complexes, and to biochemical reactions taking place in certain natural ecosystems. Activity caused by different microbial proteinases is demonstrated in agar plates at pH 3.6. The possibility of using the method for studies on proteolytic enzymes at low pH values is suggested.