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Benefits of Soleris® over the Conventional Method for Enumeration of Microbial Load in Salacia Herbal Extract

SADANAND YEWALE

SADANAND YEWALE

Herbal Division, Sava Healthcare Limited, Research CenterChinchwad, Pune, India
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YEWALE, SADANAND
, 
ZEBA FARASH

ZEBA FARASH

Analytical Development Laboratory, Sava Healthcare Limited, Research CenterChinchwad, Pune, India
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FARASH, ZEBA
, 
SANMAN KOLHE

SANMAN KOLHE

Herbal Division, Sava Healthcare Limited, Research CenterChinchwad, Pune, India
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KOLHE, SANMAN
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SASIDHARAN SAKKAN

SASIDHARAN SAKKAN

QC, Microbiology Division, Sava Healthcare Limited, KIADB Industrial AreaMalur, India
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SAKKAN, SASIDHARAN
, 
SHRINIVAS BHOPE

SHRINIVAS BHOPE

Analytical Development Laboratory, Sava Healthcare Limited, Research CenterChinchwad, Pune, India
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BHOPE, SHRINIVAS
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PRADNYA AMBEKAR

PRADNYA AMBEKAR

Neogen Food and Animal Security (India) Pvt. Ltd.Kochi, India
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AMBEKAR, PRADNYA
 y 
SRIRAM PADMANABHAN

SRIRAM PADMANABHAN

Herbal Division, Sava Healthcare Limited, Research CenterChinchwad, Pune, India
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PADMANABHAN, SRIRAM
  
27 dic 2020
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Categoría del artículo: original-paper
Publicado en línea: 27 dic 2020
Páginas: 453 - 462
Recibido: 18 jul 2020
Aceptado: 08 oct 2020
DOI: https://doi.org/10.33073/pjm-2020-048
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© 2020 Sadanand Yewale et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Schematic representation of the Soleris® system. In Step 1, an aliquot (10 g) of the herbal extract powder is weighed in a sterile container and re-suspended in 90 ml of sterile peptone water. The solution was mixed by vigorous vortexing for 2 min and further dilutions up to 10–3 were prepared by adding 1 ml of the diluted solution to 9 ml of sterile peptone and 1 ml of the dilutions were placed in the Soleris® vials in a laminar biosafety hood under sterile conditions (Step 2). In Step 3, the inoculated vials are placed in the selected drawer location in Soleris® instrument and the vials incubated at an appropriate temperature based on type of assay chosen (TVC, yeast and molds or coliforms). As organisms grow in the broth medium, the carbon dioxide (CO2) produced diffuses through a membrane layer into a soft agar plug containing a dye indicator and the change in the color of the dye is read by the Soleris® instrument (Step 4). A detection curve is generated in real time (Step 5) and the Soleris® software indicates a positive test result in less than 24 h for a sample with microbial contamination and samples producing no detection curve within 24 h are considered negative at the test threshold selected.
Schematic representation of the Soleris® system. In Step 1, an aliquot (10 g) of the herbal extract powder is weighed in a sterile container and re-suspended in 90 ml of sterile peptone water. The solution was mixed by vigorous vortexing for 2 min and further dilutions up to 10–3 were prepared by adding 1 ml of the diluted solution to 9 ml of sterile peptone and 1 ml of the dilutions were placed in the Soleris® vials in a laminar biosafety hood under sterile conditions (Step 2). In Step 3, the inoculated vials are placed in the selected drawer location in Soleris® instrument and the vials incubated at an appropriate temperature based on type of assay chosen (TVC, yeast and molds or coliforms). As organisms grow in the broth medium, the carbon dioxide (CO2) produced diffuses through a membrane layer into a soft agar plug containing a dye indicator and the change in the color of the dye is read by the Soleris® instrument (Step 4). A detection curve is generated in real time (Step 5) and the Soleris® software indicates a positive test result in less than 24 h for a sample with microbial contamination and samples producing no detection curve within 24 h are considered negative at the test threshold selected.

Fig. 2. a)

Microbial growth curve in Soleris® NF-TVC vials of various dilutions. Red curve denotes growth with sample SR011903 of10–3 dilution while blue curve denotes 10–5 dilution of the sample. Microbial detection was seen in both the dilutions.
Microbial growth curve in Soleris® NF-TVC vials of various dilutions. Red curve denotes growth with sample SR011903 of10–3 dilution while blue curve denotes 10–5 dilution of the sample. Microbial detection was seen in both the dilutions.

Fig. 2. b)

Sample RDP/SR/070/SS01 showed positive results for NF-TVC in Soleris® at 10–2 dilution (red curve) while the blue curve denotes 10–3 dilution.
Sample RDP/SR/070/SS01 showed positive results for NF-TVC in Soleris® at 10–2 dilution (red curve) while the blue curve denotes 10–3 dilution.

Fig. 2. c)

Sample RDP/SR/070/ES02 showed detection of microbes in all the dilutions in Soleris® NF-TVC vials. Blue curve denotes 10–3 dilution while the 10–2 dilution is represented by the red curve.
Sample RDP/SR/070/ES02 showed detection of microbes in all the dilutions in Soleris® NF-TVC vials. Blue curve denotes 10–3 dilution while the 10–2 dilution is represented by the red curve.

Fig. 2. d)

Sample RDP/SR/070/GR03 showed detection in all the dilutions in Soleris® NF TVC vials. Blue curve denotes 10–3 dilution while the red curve denotes 10–2 dilution.
Sample RDP/SR/070/GR03 showed detection in all the dilutions in Soleris® NF TVC vials. Blue curve denotes 10–3 dilution while the red curve denotes 10–2 dilution.

Fig. 2. e)

Sample RDP/SR/068 showed detection in all the dilutions in Soleris® NF-TVC vials. Red curve denotes 10–3 dilution while the blue curve shows growth with 10–2 dilution of the sample.
Sample RDP/SR/068 showed detection in all the dilutions in Soleris® NF-TVC vials. Red curve denotes 10–3 dilution while the blue curve shows growth with 10–2 dilution of the sample.

Fig. 2. f)

RDP/SR/134 sample showed no bacterial growth in any of the dilutions (10–3 and 10–5) tested. This is represented by blue and black flat lines respectively.
RDP/SR/134 sample showed no bacterial growth in any of the dilutions (10–3 and 10–5) tested. This is represented by blue and black flat lines respectively.

Fig. 2. g)

RDP/SR/136 sample showed microbial detection only in 10–3 dilution in NF-TVC Soleris® vials denoted by the red curve. The blue flat line denotes no growth seen with 10–5 dilution of this sample.
RDP/SR/136 sample showed microbial detection only in 10–3 dilution in NF-TVC Soleris® vials denoted by the red curve. The blue flat line denotes no growth seen with 10–5 dilution of this sample.

Fig. 3. a)

Sample SR011903 showed microbial detection in 10–1 dilution in Soleris® DYM 109-C vials represented by blue curve. Black curve denotes 10–3 dilution while the red curve denotes 10–2 dilution, both of which did not show any growth.
Sample SR011903 showed microbial detection in 10–1 dilution in Soleris® DYM 109-C vials represented by blue curve. Black curve denotes 10–3 dilution while the red curve denotes 10–2 dilution, both of which did not show any growth.

Fig. 3. b)

Sample RDP/SR/136 showed microbial detection in 10–1 dilution for DYM 109-C Soleris® vials represented by the red curve.
Sample RDP/SR/136 showed microbial detection in 10–1 dilution for DYM 109-C Soleris® vials represented by the red curve.

Soleris® results for the total yeast and mold counts_

S.N.Sample IDDilutions inoculatedGrowth/No growthDetection time (h)
1SR011903101Growth34.4
102No Growth–
103No Growth–
2RDP/SR/070/SS01 (Steam sterilized SR011903)101No Growth–
102No Growth–
103No Growth–
3RDP/SR/070/ES02 (ETO treated SR011903)101No Growth–
102No Growth–
103No Growth–
4RDP/SR/070/GR03 (Gamma irradiated SR011903)101No Growth–
102No Growth–
103No Growth–
5RDP/SR/068 (anti-fungal agent treated SR011903)101No Growth–
102No Growth–
103No Growth–
6RDP/SR/134 (anti-bacterial agent treated SR011903)101No Growth–
102No Growth–
103No Growth–
7RDP/SR/136 (commercial Salacia extract)101Growth26.2
102No growth–
103No growth–

Soleris® results for total aerobic microbial count_

S.N.Sample IDDilutions inoculatedGrowth/No growthDetection time (h)
1SR011903103105GrowthGrowth2.74.8
2RDP/SR/070/SS01102103GrowthNo growth11.4ND
3RDP/SR/070/ES02102103GrowthGrowth12.211.9
4RDP/SR/070/GR03102103GrowthGrowth15.119.9
5RDP/SR/068102103GrowthGrowth10.612.3
6RDP/SR/134103105No growthNo growthNo detectionNo detection
7RDP/SR/136102103GrowthNo growth13.1No detection

Comparison of a total aerobic count between Soleris® (NF-TVC) and the conventional method (Salacinol content)_

S.N.Sample IDTotal aerobic count by Soleris® (CFU/g)*Total aerobic count by plating in Soleris® medium (CFU/g)**Conventional plating method***RLOD#Salacinol content (%)
1SR011903 (Control)> 100,000109,000> 1051.090.52
2RDP/SR/070/SS01 (Steam sterilized SR011903)< 1,0004903501.40ND
3RDP/SR/070/ES02 (ETO treated SR011903)> 1,0001,1002,7000.41ND
4RDP/SR/070/GR03 (Gamma irradiated SR011903)< 1,0002808900.31ND
5RDP/SR/068 (anti-fungal agent treated SR011903)> 1,0001,6001,5001.060.506
6RDP/SR/134 (anti-bacterial agent treated SR011903)> 1010200.500.48
7RDP/SR/136 (commercial Salacia extract)> 1,0001,5002,3000.650.059

Comparison between Soleris® (DYM-109) and the conventional method for the total yeasts and molds_

S.N.Sample IDSoleris® (CFU/g)Conventional method (CFU/g)Acceptable/Not acceptable
1SR011903> 10380  Not acceptable
2RDP/SR/070/SS01< 10< 10  Acceptable
3RDP/SR/070/ES02< 10< 10  Acceptable
4RDP/SR/070/GR03< 10< 10  Acceptable
5RDP/SR/068< 10< 10  Acceptable
6RDP/SR/134< 10< 10  Acceptable
7RDP/SR/136> 10290  Not acceptable
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