High-throughput farming of animals for an essential purpose such as large scale health and production of hogs is a challenge for the food industry in the modern world. The problem is that the breeding of livestock for fast growth or high yields of meat is often associated with illness and microbial infection that develop under the breeding conditions. Piglet diarrhea is most common pig disease, leading to heavy mortality and thereby economic loss. We proved that chemical drugs can relieve the symptoms of diarrhea in ill piglets, but they do not treat the underlying cause, i.e. significantly altered bacterial gut flora. Using Illumina sequencing of fecal DNA, we showed that the bacterial gut flora of piglets treated with antibiotics remain close to the ill conditions. However, using Illumina sequencing of fecal DNA from piglets treated with a specific Bacillus (Bacillus subtilis Y-15, B. amyloliquefaciens DN6502 and B. licheniformis SDZD02) demonstrated the efficiency of natural bioproducts not only on curing diarrhea, but also on beneficial bacteria to re-establish in the piglet gut. We therefore propose a new natural “medicine” to be explored by the world farm animal agriculture industry, particularly for sustainable improvement of swine livestock production and health.
We parsed the microbial genome database using Bombyx mori chemosensory proteins (BmorCSPs) as templates. We extracted eleven bacterial CSPs (B-CSPs) from various microorganisms such as Kitasatospora griseola, K. purpeofusca, K. CB01950, K. MBT66, Escherichia coli, Macrococcus caseolyticus and Acinetobacter baumannii, a known infectious prokaryotic symbiont of various insects, particularly the human body louse. We then parsed the body louse Pediculus humanis corporis genomic database for CSPs. We found six P. humanis corporis (Phum) CSPs all grouped in the same gene cluster. Sequence alignment, structure modeling and phylogenetic analysis of CSP proteins in bacteria and insects reveal duplication, conservation, gene loss, but also diversification and neofunctionalization that took place at a common stage in this ancestral gene family. Phylogenetic analysis of the amino acid sequences also reveals association of CSPs with other prokaryotic gene families, mainly enzymes and secondary metabolites transporters. Their ability to bind lipids and their proved existence and diversity in infectious bacterial prokaryote systems strongly argue for some important general functions in the cellular metabolism process.
The relationship between salt bridges and stability/enzymatic activity is unclear. We studied this relationship by systematic alanine-scanning mutation analysis using the typical M4 family metalloprotease Pseudomonas aeruginosa elastase (PAE, also known as pseudolysin) as a model. Structural analysis revealed seven salt bridges in the PAE structure. We constructed ten mutants for six salt bridges. Among these mutants, six (Asp189Ala, Arg179Ala, Asp201Ala, Arg205Ala, Arg245Ala and Glu249Ala) were active and four (Asp168Ala, Arg198Ala, Arg253Ala, and Arg279Ala) were inactive. Five mutants were purified, and their catalytic efficiencies (k cat/K m), half-lives (t 1/2) and thermal unfolding curves were compared with those of PAE. Mutants Asp189Ala and Arg179Ala both showed decreased thermal stabilities and increased activities, suggesting that the salt bridge Asp189-Arg179 stabilizes the protein at the expense of catalytic efficiency. In contrast, mutants Asp201Ala and Arg205Ala both showed slightly increased thermal stability and slightly decreased activity, suggesting that the salt bridge Asp201-Arg205 destabilizes the protein. Mutant Glu249Ala is related to a C-terminal salt bridge network and showed both decreased thermal stability and decreased activity. Furthermore, Glu249Ala showed a thermal unfolding curve with three discernable states [the native state (N), the partially unfolded state (I) and the unfolded state (U)]. In comparison, there were only two discernable states (N and U) in the thermal unfolding curve of PAE. These results suggest that Glu249 is important for catalytic efficiency, stability and unfolding cooperativity. This study represents a systematic mutational analyses of salt bridges in the model metalloprotease PAE and provides important insights into the structure-function relationship of enzymes.
We deal with various strains of Lactobacillus that can maintain the intestinal microbiome of rats treated with cyclophosphamide, an anticancer agent (chemotherapy). We use MiSeq and various types of statistical tests to prove that cyclophosphamide in rats alters the intestinal microbiome, favoring the growth of various fungi that are extremely harmful to intestinal metabolism. On the contrary, when Lactobacillus 3L is administered together with cyclophosphamide, we prove that the microbiome is preserved by having a much better intestinal metabolism.
Background We examined the impact of using a probiotic containing three different Lactobacilli (3L) on the gut microbiome of rats following cyclophosphamide (CTX) treatment. CTX corresponded to chemotherapy which is used for human cancer treatment and known to have adversive effects on the immune system. Methods We conducted our experiment with ten rats in five different experimental groups which included control, CTX treated, and then low, medium, and high probiotic treatment with CTX treatment. Of these ten rats in each group, we sequenced the stool of three of them using both ITS and 16S sequencing. We then went on to examine the taxonomic composition of these samples to determine whether probiotic treatment helped the rat’s microbiome return to similar structure as the control rats. Results We used Illumina MiSeq sequencing to generate sequencing data from microbial genomic DNA libraries, which is useful for testing the effects of 3L on bacteria and fungi. Microbiome analysis, phylogenetic and classification reports, and community data have all backed up the experiments and findings that 3L had a significant positive impact on the microbiome. Furthermore, the effect on specific metabolic pathways aids in deriving the study’s conclusion (use of 3L in chemotherapy) to the mode of action, mechanistically by correcting microbiota composition and enhancing specific gut metabolic functions. Conclusions Through experimental results using an in vivo model, we suggested the role of novel natural probiotics 3L, 3 Lactobacilli in the establishment of a strong and sustainable beneficial healthy gut flora, after CTX chemotherapy. We suggested some new adjuvants to chemotherapy as drugs + lactobacillus treament using the rat CTX model (immunosuppression caused by cyclophosphamide). Furthermore, in numerous studies that reported the use of probiotics involving Lactobacillus in post-chemo or post-surgical procedures, we proposed a new probiotic formulation (L. acidophilus + L. casei + L. plantarum) to be further studied and explored in the prevention of health condition loss by alteration of the general immune system.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.