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New methods for isolation of keratolytic bacteria inducing intractable hoof wall cavity ( Gidoh ) in a horse; double screening procedures of the horn powder agar-translucency test and horn zymography

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ABSTRACT

To establish a new system to isolate keratolytic bacteria from the hoof wall cavity (Gidoh) of a racehorse, we invented the horn powder agar-translucency(HoPAT) test and horn zymography (HZ). Using routine bacteriological techniques and these methods, we isolated five strains of keratolytic soil bacteria, whichwere then identified by means of 16S ribosomal RNA (rRNA) gene sequencing analysis. The findings from the study on the horse suggested that Brevibacteriumluteolum played the main role in the local fragility of the hoof, eventually forming a Gidoh in coordination with four other strains of keratolytic bacteria.The double screening procedures of the HoPAT test and HZ were useful and easy techniques for isolating the keratolytic bacteria from the horn lesions.

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In the horn zymography, the keratolytic activity of each isolated bacterium (from No. 1 to No. 5) was visible as one to four white bands in thewater-soluble horn-combined acrylamide gel stained with Coomassie Brilliant Blue (CBB) in the secondary screening examination.
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fig_004: In the horn zymography, the keratolytic activity of each isolated bacterium (from No. 1 to No. 5) was visible as one to four white bands in thewater-soluble horn-combined acrylamide gel stained with Coomassie Brilliant Blue (CBB) in the secondary screening examination.

Mentions: One of the culture results from the horn powder agar-translucency test: The strain in (corresponding to No. 2) appeared as a yellowish colony and presented a stronger and wider translucency characteristic around the colony on the horn powderagar than other strains. The colony in on the right side of No. 2 showed weak translucency,whereas had no translucency characteristic.


New methods for isolation of keratolytic bacteria inducing intractable hoof wall cavity ( Gidoh ) in a horse; double screening procedures of the horn powder agar-translucency test and horn zymography
In the horn zymography, the keratolytic activity of each isolated bacterium (from No. 1 to No. 5) was visible as one to four white bands in thewater-soluble horn-combined acrylamide gel stained with Coomassie Brilliant Blue (CBB) in the secondary screening examination.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5383627&req=5

fig_004: In the horn zymography, the keratolytic activity of each isolated bacterium (from No. 1 to No. 5) was visible as one to four white bands in thewater-soluble horn-combined acrylamide gel stained with Coomassie Brilliant Blue (CBB) in the secondary screening examination.
Mentions: One of the culture results from the horn powder agar-translucency test: The strain in (corresponding to No. 2) appeared as a yellowish colony and presented a stronger and wider translucency characteristic around the colony on the horn powderagar than other strains. The colony in on the right side of No. 2 showed weak translucency,whereas had no translucency characteristic.

View Article: PubMed Central - PubMed

ABSTRACT

To establish a new system to isolate keratolytic bacteria from the hoof wall cavity (Gidoh) of a racehorse, we invented the horn powder agar-translucency(HoPAT) test and horn zymography (HZ). Using routine bacteriological techniques and these methods, we isolated five strains of keratolytic soil bacteria, whichwere then identified by means of 16S ribosomal RNA (rRNA) gene sequencing analysis. The findings from the study on the horse suggested that Brevibacteriumluteolum played the main role in the local fragility of the hoof, eventually forming a Gidoh in coordination with four other strains of keratolytic bacteria.The double screening procedures of the HoPAT test and HZ were useful and easy techniques for isolating the keratolytic bacteria from the horn lesions.

No MeSH data available.