Manipulation of Lactobacillus Probiotic Strains to Produce Heterologous Beta-Glucanase for Chickens

Application of enzymes as feed additives is common in the livestock industry, especially in poultry, to eliminate the antinutritional factors present in the diets of chickens. However, the efficiency of enzymes seldom achieves their desired effects because of destruction during feed processing and unsuitable conditions in the gastrointestinal tract. Thus, in the present study, investigations were carried out to evaluate the potential of 12 Lactobacillus strains as delivery vehicles for a heterologous β-glucanase enzyme in poultry. The 12 Lactobacillus strains used were L. crispatus I12, L. acidophilus I16 and I26, L. fermentum I24, I25, C16 and C17, and L. brevis I23, I211, I218, C1 and C10. The strains were found to exhibit resistance to chloromphenicol, erythromycin and tetracycline in varying degrees. The erythromycin resistance of L. acidophilus I16 and I26, and L. fermentum I24 and C17 could be cured by using novobiocin, and L. brevis C10 cured by using acriflavin. The chloromphenicol and tetracycline resistances of all the resistant strains were not eliminated even after prolonged curing in sublethal concentrations of individual or mixtures of curing agents such as novobiocin, ethidium bromide, acriflavin or SDS. Electrotransformation efficiency of the Lactobacillus strains was affected by growth phase, growth and recovery medium, cell density, electroporation buffer, buffer strength, plasmid concentration and electrical pulse. At optimized conditions, the strains were transformed at 103-104 transformants/μg plasmid DNA. The erythromycin susceptible wild-type strains (L. crispatus I12, L. brevis I23, I211 and I218, and L. fermentum I25) and cured derivatives (L. acidophilus I16C and I26C, L. brevis v C10C, and L. fermentum I24C and C17C) were then transformed at optimized conditions with plasmid pSA3b6, which carried a β-glucanase gene from Bacillus amyloliquefaciens. Five wild-type Lactobacillus strains, namely, L. crispatus I12, L. fermentum I25, L. brevis I23, I211 and I218 and a cured derivative, L. brevis C10C, which could retain the plasmid at a comparatively higher rate, were used for subsequent studies. The Lactobacillus transformants were found to secrete 32-52 U/ml of β- glucanase. Optimum activity of the enzyme was at 39 oC and pH 5-6. A loss of 0.4-1.6 U/generation of β-glucanase was observed when the strains were grown under nonselective pressure. PCR analyses of gastrointestinal samples of chickens fed transformed Lactobacillus strains revealed that the strains could not persist for more than 24 h in the gut. The β- glucanase activity detected in the jejunum and ileum of chickens fed transformed Lactobacillus strains was found to be 2-9.4 folds higher than those obtained from other intestinal sites. In the feeding trial, supplementation of transformed Lactobacillus strains to chickens significantly (P<0.05) improved the body weight by 2.5 %, and the feed conversion ratio by 1.0-2.6 %. In addition, the apparent metabolizable energy, digestibilities of crude protein and dry matter of feed were improved by 3.4 %, 5.9 % and 3.5 %, respectively. The intestinal fluid viscosity was reduced by 21-46 %. The relative weights of organs and intestinal segments (pancrease, liver, duodenum, jejunum, ileum, cecum and colon) were also reduced by 6-27 %, and the relative length of intestinal segments (duodenum, jejunum, ileum and cecum) was reduced by 8-15 %. Histological examination of the intestinal tissues showed that the jejunal villus height of chickens fed diet supplemented with transformed Lactobacillus strains was significantly (P<0.05) higher than those of chickens fed other dietary treatments. The transformed Lactobacillus strains were also found to reduce the time of feed passage rate by 2.2 h. vi The results of the present study showed that the Lactobacillus strains have the potential to be used as delivery vehicles for a heterologous β-glucanase enzyme in poultry

Estimation of 16S RNA gene copy number in several probiotic Lactobacillus strains isolated from the gastrointestinal tract of chicken.

The copy numbers of 16S rRNA genes in 12 probiotic Lactobacillus strains of poultry origin were analyzed. Genomic DNA of the strains was digested with restriction endonucleases that do not cut within the 16S rRNA gene of the strains. This was followed by Southern hybridization with a biotinylated probe complementary to the 16S rRNA gene. The copy number of the 16S rRNA gene within a Lactobacillus species was found to be conserved. From the hybridization results, Lactobacillus salivarius I 24 was estimated to have seven copies of the 16S rRNA gene, Lactobacillus panis C 17 to have five copies and Lactobacillus gallinarum strains I 16 and I 26 four copies. The 16S rRNA gene copy numbers of L. gallinarum and L. panis reported in the present study are the first record. Lactobacillus brevis strains I 12, I 23, I 25, I 211, I 218 and Lactobacillus reuteri strains C 1, C 10, C 16 were estimated to have at least four copies of the 16S rRNA gene. In addition, distinct rRNA restriction patterns which could discriminate the strains of L. reuteri and L. gallinarum were also detected. Information on 16S rRNA gene copy number is important for physiological, evolutionary and population studies of the bacteria

L\u27Apres Midi d\u27une...

A picnic was spread on the bank and they surrounded it in positions of spiritual abandon. One, a long and somewhat undulant girl, lay among them gazing into the river that rolled peacefully yet dense with clay after the spring flood. The opposite bank, mysterious as an arabesque, hid in its shadow-work of trees the lives that like theirs, had moved back from afternoon; lives bound to the waterway by probing beak and wading leg, by a diet of fish or of insects Whose larval time is passed in mud and water. Look closely! implied the arabesque: a leaf may become a bird

Numb Chin Syndrome: Atypical presentation of metastatic breast cancer

Introduction/Background: Numb chin syndrome (NCS) is an uncommon but known complication of cancer. Prompt recognition is imperative to directed treatment. In some instances, numb chin may be complicated by other findings. &nbsp; Case Report: A 53-year-old female with past medical history of estrogen receptor positive invasive ductal breast carcinoma underwent partial mastectomy, chemotherapy, and radiation. After 6 years remission, new lung and liver metastases occurred, and chemotherapy resumed. &nbsp;Imaging was negative.&nbsp; Eight weeks later, she developed left chin numbness followed by progressive difficulty retaining food and drink.&nbsp; Her left lower lip weakened. &nbsp; Neurological examination revealed left lower lip weakness (showing a “droop” with attempted smile). &nbsp;The area of decreased sensation to sharp touch had expanded from the left chin to the nasolabial fold.&nbsp; The remainder of her neurological examination was unremarkable. Contrast-enhanced MRI was repeated which revealed a 1.9 cm enhancing left parotid mass. &nbsp; Summary/Conclusion: NCS is defined as an ipsilateral loss of chin sensation. If there is no history of trauma or dental injury and especially if the presentation is atypical, then cancer is the most likely etiology. This case began as an uncomplicated NCS that rapidly expanded to involve a greater portion of the trigeminal nerve distribution, in addition to a branch of the facial nerve.&nbsp; Thus, each focal deficit was not the result of separate distal nerve lesions, but rather partial proximal cranial neuropathies at their parotid gland propinquity

The Chin as a Domain Widener in American Sign Language (ASL)

In this paper, we investigate a grammaticalized facial expression in American Sign Language (ASL) called flat chin, which we propose functions as a general-purpose domain widener, targeting both quantificational domains as well as the scales used by gradable predicates. Our analysis allows for flat chin to target nearly any expression involving a domain, and is based on Morzycki's (2012) analysis of extreme degree modifiers. Such an analysis both expands our understanding of ASL as well Language more broadly, as few general-purpose domain wideners have yet been reported and fewer still are reported to occur as a non-manual (non-hand) markers in a sign language

Usability of Food and Beverage Packs in Hospital - Experiences from the Renal Ward

Abstract presented at The 21st IAPRI World Conference on Packaging, 19-22 June 2018, Zhuhai, Chin

Flavor Asymmetry of Nucleon Sea from Detailed Balance

In this study, the proton is taken as an ensemble of quark-gluon Fock states. Using the principle of detailed balance, we find $\bar{d}-\bar{u} \approx 0.124$, which is in surprisingly agreement with the experimental observation.Comment: 4 pages and no figures. Talk presented at the Third Circum-Pan-Pacific Symposium on "High Energy Spin Physics", Oct. 8-13, 2001, Beijing, Chin

Nonsurgical genioplasty

Chin contributes to facial balance and harmony. Appropriate treatment of aesthetic deformities of the will often improves the appearance of the mouth, lips, and nose. Augmentation of the chin can be performed with injectable fillers or autologous fat, placement of an alloplastic chin implant, or with chin's bony osteotomy. Determining the best procedure for a patient requires careful consideration of his anatomy, as well as the risks and benefits of each treatment. Fillers offer a nonsurgical, nonpermanent method of correction of chin retrusion in the appropriate patient