Characterization and Electrotransformation of Lactobacillus Crispatus Isolated from Chicken Crop and Intestine

Beasley, Shea; Takala, Timo M.; Reunanen, Justus; Apajalahti, Juha; Saris, Per E. J.

doi:10.1093/ps/83.1.45

Cited by 32 publications

(26 citation statements)

References 25 publications

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“…Enhancing the maturation of the crop microbiota after hatch may provide a less susceptible environment to pathogenic load, and appropriate probiotics (Beasley et al, 2004;Abbas Hilmi et al, 2007), and carbohydrates and other nutrients targeted for lactobacilli may be useful to accomplish that goal (Lebeer et al, 2011). Although in ovo application of probiotics would seem appropriate to ensure earliest possible bacterial colonisation (Edens et al, 1997), the authors are unaware of successful applications.…”

Section: Developing and Maintaining A Healthy Cropmentioning

confidence: 99%

The role of the crop in poultry production

Classen

Apajalahti

Svihus

et al. 2016

World's Poultry Science Journal

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The importance of the crop is often underestimated in poultry production. In addition to storing ingested feed, it also can impact nutrient digestion by digesta softening and the initial activity of feed (endogenous and exogenous) and microbial enzymes. The crop represents the first major defence against poultry pathogens and zoonotic organisms with well established adaptive and innate immune function, and a lactobacilli dominated microbiota capable of reducing the passage of these organisms further along the digestive tract. However, the potential to improve bird productivity and health, as well as affect meat and egg safety, are influenced by the nature of the diet, and in particular feed entry and extended presence in the crop. This is required to promote lactobacilli fermentation, the production of lactic acid and other volatile fatty acids, and the lowering of crop pH. Management practices such as meal feeding and the use of lighting programs with extended dark periods encourage crop utilisation. Further, the use of feed additives such as prebiotics and probiotics may enhance crop function, which in turn contributes to well-being of the entire digestive tract. A healthy and functional crop, along with other segments of digestive tract, has increased importance in an era of reduced antibiotic use in poultry feeds.

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Section: Developing and Maintaining A Healthy Cropmentioning

confidence: 99%

The role of the crop in poultry production

Classen

Apajalahti

Svihus

et al. 2016

World's Poultry Science Journal

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“…Electrocompetent E. coli MC1061 and TOP10 were prepared and transformed according to standard methods (32). Lactobacillus acidophilus and Lactobacillus gasseri were transformed using the method of Luchansky et al (33), while Lactobacillus gallinarum was transformed using the method of Beasley et al (34).…”

Section: Methodsmentioning

confidence: 99%

Evaluation of Phytate-Degrading Lactobacillus Culture Administration to Broiler Chickens

Askelson

Campasino

Lee

et al. 2014

Appl Environ Microbiol

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Probiotics have been demonstrated to promote growth, stimulate immune responses, and improve food safety of poultry. While widely used, their effectiveness is mixed, and the mechanisms through which they contribute to poultry production are not well understood. Microbial phytases are increasingly supplemented in feed to improve digestibility and reduce antinutritive effects of phytate. The microbial origin of these exogenous enzymes suggests a potentially important mechanism of probiotic functionality. We investigated phytate degradation as a novel probiotic mechanism using recombinant Lactobacillus cultures expressing Bacillus subtilis phytase. B. subtilis phyA was codon optimized for expression in Lactobacillus and cloned into the expression vector pTRK882. The resulting plasmid, pTD003, was transformed into Lactobacillus acidophilus, Lactobacillus gallinarum, and Lactobacillus gasseri. SDS-PAGE revealed a protein in the culture supernatants of Lactobacillus pTD003 transformants with a molecular weight similar to that of the B. subtilis phytase. Expression of B. subtilis phytase increased phytate degradation of L. acidophilus, L. gasseri, and L. gallinarum approximately 4-, 10-, and 18-fold over the background activity of empty-vector transformants, respectively. Phytase-expressing L. gallinarum and L. gasseri were administered to broiler chicks fed a phosphorusdeficient diet. Phytase-expressing L. gasseri improved weight gain of broiler chickens to a level comparable to that for chickens fed a control diet adequate in phosphorus, demonstrating proof of principle that administration of phytate-degrading probiotic cultures can improve performance of livestock animals. This will inform future studies investigating whether probiotic cultures are able to provide both the performance benefits of feed enzymes and the animal health and food safety benefits traditionally associated with probiotics. Lactobacillus species are important inhabitants of the gastrointestinal tracts of humans and animals and are increasingly being used as probiotic microorganisms due to their health-promoting properties (1, 2). Probiotics, sometimes called direct-fed microbials (DFM) when used in animals (3), are live microorganisms administered to confer a health benefit upon the host (4). Administration of probiotic Lactobacillus to poultry has been demonstrated to promote growth at levels similar to antibiotics (5, 6) and to reduce gastrointestinal colonization of human foodborne pathogens, including Campylobacter (7, 8), Clostridium (9), and Salmonella (10, 11). Because of concern over antibiotic-resistant pathogens and pressure from both consumers and regulatory agencies, probiotics have received increased interest as potential alternatives to antibiotic growth promoters (12). While probiotics are used widely in livestock production (13), their effectiveness is varied, and the mechanisms responsible for their benefits are not well understood.Phosphorus is an essential nutrient in poultry production (14), with dietary deficiencies leadin...

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“…Bacillus anthracis (85) lux Citrobacter rodentium (23) lux Brucella melitensis (86) lux Bifidobacterium breve (87) lux Escherichia coli (21) lux, luc Listeria monocytogenes (20) lux Mycobacterium tuberculosis (88) lux, luc Pseudomonas aeruginosa (89) lux Pseudomonas fluorescence (90) lux Salmonella enterica Typhimurium (75) lux Staphylococcus aureus (19,29) lux Streptococcus pneumonia (30) lux stresses to host organisms. 32 The mCherry fluorophore is also highly resistant to photo bleaching, which means that the fluorophore will not lose its ability to fluoresce during continuous illumination.…”

Section: Speciesmentioning

confidence: 99%

“…Added to the challenge are the differences in transformation efficiency obtained for strains of the same species, as demonstrated by using the same electroporation protocol on different strains of L. crispatus. 74,75 Reporter genes are usually cloned into plasmids or integrated into the genome. By integration into the genome, gene expression is much more stable and antibiotic selection is not required, which is a big advantage in animal studies.…”

Section: Expression Of Reporter Genes In Lactic Acid Bacteriamentioning

confidence: 99%

Reporter systems forin vivotracking of lactic acid bacteria in animal model studies

2015

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Bioluminescence (BLI) and fluorescence imaging (FI) allow for non-invasive detection of viable microorganisms from within living tissue and are thus ideally suited for in vivo probiotic studies. Highly sensitive optical imaging techniques detect signals from the excitation of fluorescent proteins, or luciferase-catalyzed oxidation reactions. The excellent relation between microbial numbers and photon emission allow for quantification of tagged bacteria in vivo with extreme accuracy. More information is gained over a shorter period compared to traditional pre-clinical animal studies. The review summarizes the latest advances in in vivo bioluminescence and fluorescence imaging and points out the advantages and limitations of different techniques. The practical application of BLI and FI in the tracking of lactic acid bacteria in animal models is addressed.

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Characterization and Electrotransformation of Lactobacillus Crispatus Isolated from Chicken Crop and Intestine

Cited by 32 publications

References 25 publications

The role of the crop in poultry production

The role of the crop in poultry production

Evaluation of Phytate-Degrading Lactobacillus Culture Administration to Broiler Chickens

Reporter systems forin vivotracking of lactic acid bacteria in animal model studies

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