Overview of cloning in lactic acid bacteria: Expression and its application of probiotic potential in inflammatory bowel diseases.

Inflammatory bowel disease (IBD) imposes a significant impact on the quality of life for affected individuals. However, there was a current lack of a systematic summary regarding the latest epidemic trends and the underlying pathogenesis of IBD. This highlights the need for a thorough examination of both the epidemiological aspects of IBD and the specific mechanisms by which lactic acid bacteria (LAB) contribute to mitigating this condition. In developed countries, higher incidences and death rates of IBD have been observed, influenced by a combination of environmental and genetic factors. LAB offer significant advantages and substantial potential for enhancing IBD treatment. LAB's capabilities include the production of bioactive metabolites, regulation of gut immunity, protection of intestinal mechanical barriers, inhibition of oxidative damage, and restoration of imbalanced gut microbiota. The review suggests that screening effective LAB using cell models and metabolites, optimizing LAB intake through dose-effect studies, enhancing utilization through nanoencapsulation and microencapsulation, investigating mechanisms to deepen the understanding of LAB, and refining clinical study designs. These efforts aim to contribute to comprehending the epidemic trend, pathogenesis, and treatment of IBD, ultimately fostering the development of targeted therapeutic products, such as LAB-based interventions.

Antitoxin activity of aqueous extract of Cyclea peltata root against Naja naja venom.

OBJECTIVES: Snakebites are a significant and severe global health problem. Till date, anti-snake venom serum is the only beneficial remedy existing on treating the snakebite victims. As antivenom was reported to induce early or late adverse reactions to human beings, snake venom neutralizing potential for Cyclea peltata root extract was tested for the present research by ex vivo and in vivo approaches on Naja naja toxin. MATERIALS AND METHODS: Ex vivo evaluation of venom toxicity and neutralization assays was carried out. The root extracts from C. peltata were used to evaluate the Ex vivo neutralization tests such as acetylcholinesterase, protease, direct hemolysis assay, phospholipase activity, and procoagulant activity. Gas chromatography-mass spectrometry (GC-MS) analysis from root extracts of C. peltata was done to investigate the bioactive compounds. RESULTS: The in vivo calculation of venom toxicity (LD50) of N. naja venom remained to be 0.301 µg. C. peltata root extracts were efficiently deactivated the venom lethality, and effective dose (ED50) remained to be 7.24 mg/3LD50 of N. naja venom. C. peltata root extract was found effective in counteracting all the lethal effects of venom. GC-MS analysis of the plant extract revealed the presence of antivenom compounds such as tetradecanoic and octadecadienoic acid which have neutralizing properties on N. naja venom. CONCLUSION: The result from the ex vivo and in vivo analysis indicates that C. peltata plant root extract possesses significant compounds such as tetradecanoic acid hexadecanoic acid, heptadecanoic acid, and octadecadienoic acid which can counteract the toxins present in N. naja.

Development of Chicken Egg Yolk Antibodies against Streptococcus mitis - Purification and Neutralizing Efficacy.

Chicken Egg Yolk antibodies (IgY) were raised in 24 week old white leg horn chickens against Streptococcus mitis (MTCC 2696). The chickens received Booster injections of increasing concentrations of antigen to raise the antibody level in egg yolk. The antibodies were purified from immunized chicken egg yolk by Poly ethylene Glycol (PEG) and Ammonium sulphate precipitation method and further purified by DEAE cellulose ion exchange column chromatography. High titre of more than 1:10000 antibodies were detected by Indirect antigen capture ELISA at 150(th) day of observation. IgY concentration varied in the range of 0.85 - 7.6mg/ml of yolk throughout the immunization period. Growth inhibition assay showed the absence of growth when the specific egg yolk antibodies was added to the Streptococcus mitis culture. Inhibition ELISA shows decrease in absorbance with increasing concentration of IgY. The results indicate that antibodies generated in chicken could be used for diagnosis and therapeutic purposes in case of Streptococcus mitis.

Neutralization of the pharmacological effects of Cobra and Krait venoms by chicken egg yolk antibodies.

Five-month-old white leghorn chickens were immunized with 50 microg of Common Cobra (Naja naja) and 30 microg of Krait venoms (Bungarus caeruleus) to generate antivenom antibodies against the venom antigen. Chickens received booster doses of increasing concentrations of venom at 14 days time intervals to raise the antivenom level in egg yolk. The antivenom from immunized chicken egg yolk was extracted by polyethylene glycol (PEG) and ammonium sulphate precipitation method which was further purified by DEAE cellulose ion exchange column chromatography. A high molecular weight protein of 180 kDa was detected by electrophoretic analysis which shows the purity of antivenom generated in chicken. Antibodies generated were specific and sensitive to the venom antigen. Various pharmacological activities of Cobra and Krait venoms were carried out by both in-vivo and in-vitro methods. The neutralization of lethality, hemorrhagic, edema, PLA(2) and procoagulant activity was evaluated in assays involving pre-incubation of venom and antivenom prior to testing. The antivenom was effective in neutralizing the toxic and enzymatic activities of venom. The LD(50) of venom for 18 g of mice was found to be 10 microg for Cobra and 3 microg for Krait venoms. The median effective dose (ED(50)) of anti-Cobra venom was 4.48 mg/5LD(50) and 1.0 ml neutralized 0.127 mg of Cobra venom and the median effective dose (ED(50)) of anti-Krait venom was 3.18 mg/5LD(50) and 1.0 ml neutralized 0.051 mg of Krait venom. The results indicate that antivenom generated in chicken could be used for therapeutic purposes in case of snakebite envenomation.

Studies on pharmacological effects of Russell's viper and Saw-scaled viper venom and its neutralization by chicken egg yolk antibodies.

Antivenom antibodies were raised in 24-week-old white leghorn chickens against hemotoxic venoms of Russell's viper and Saw-scaled viper snakes. Booster injections of increasing concentrations of venom were given at 14days of time interval to raise the antivenom level in egg yolk. Antibodies were extracted from immunized chicken egg yolk by Polson et al. (Polson A., Von Wechmar M.B., Van Regenmortel M.H.V. Isolation of viral IgY antibodies from yolks of immunized hens. Immunological Communications 1980; 9:475-493.) and further purified by DEAE cellulose ion exchange column chromatography, which gave pure (180-200kDa) specific antibodies against venom. High titre of more than 1:10,000 antibodies were detected by ELISA at the 135th day of observation. The lethal toxicity and various pharmacological activities like hemorrhagic activity, phospholipase activity, edema and procoagulant activities of venom were carried out by both in vivo and in vitro methods. The effectiveness of antivenom in neutralizing these effects was carried out involving pre-incubation type experiments. The median effective dose (ED50) for Russell's viper venom was 0.96mg/2LD50/18g mice and for Saw-scaled viper venom it was 1.28mg/2LD50/18g mice. One millilitre of specific antivenom was effective in neutralizing 0.110mg of Russell's viper and 0.137mg of Saw-scaled viper venoms respectively (PD50). Antivenom was effective in neutralization assays in a dose dependent manner. The results indicate that antibodies raised in chicken could effectively neutralize the pharmacological effects induced by venoms and chickens therefore present an alternative and cheaper source of specific antibody generation.