scholarly journals Proteolytic Bacillus spp. associated with tannery industries: Conventional and molecular identification

2018 ◽  
Vol 44 (4) ◽  
pp. 557-564
Author(s):  
Mihir Lal Saha ◽  
Tahsin Khan ◽  
Md Mostafa Kamal ◽  
Mohammad Nurul Islam
Keyword(s):  
Molecular Identification ◽  
Skim Milk ◽  
Bacterial Count ◽  
Subtilis Strain ◽  
Bacillus Spp ◽  
Proteolytic Activities ◽  
Glucose Agar ◽  
Colonial Morphology ◽  
Good Number ◽  
Peptone Yeast Extract Glucose

Samples collected from different stages of the tannery industry were found to be alkaline (pH 7.52 to 12.11). A good number of bacteria were found to be associated with the samples. The bacterial count ranged in between 1.34×105 to 3.44×105 cfu/ml and 1.04×105 to 6×105 cfu/ml on nutrient agar (NA) and peptone yeast extract glucose agar (PYG) medium, respectively. The maximum bacterial count was observed in bating stage while the minimum count was in the deliming stage. Primarily, 70 isolates were selected based on their different colonial morphology. After heat shock test 27 isolates were finally selected for identification and proteolytic potential. All the selected isolates were the members of different species of the genus Bacillus. The conventionally identified species were B. stearothermophilus (9), B. subtilis (4), B. brevis (3), B. pumilus (3), B. alcalophilus (2), B. badius (2), B. firmus (2) and B. lentus (2). Four important proteolytic isolates of Bacillus were selected for molecular identification. The isolates were confirmed as Bacillus subtilis strain B20 (L/P/2/1), B. subtilis strain PB18 (D/P/3/1), Bacillus sp. strain BVC38 (D/P/3′/2) and B. amyloliqefaciens strain Egy25 (B/N/3′/1). Except B/N/3′/1 all the conventional identification was in accordance with the molecular identification as the isolate B/N/3′/1 was conventionally identified as B. pumilus (B/N/3′/1). All the isolates showed positive proteolytic activities on skim milk agar and the zone ratio was in between 2.61 ± 0.44 and 6.42 ± 0.95. These isolates could be commercially utilized in the tannery and detergent industries for their proteolytic activity.

scholarly journals Bacteria Associated with the Tannery Effluent and their Alkaline Protease Activities

2012 ◽  
Vol 21 (1) ◽  
pp. 53-61 ◽  
Author(s):  
Mihir Lal Saha ◽  
K. J. M. Hashina Begum ◽  
Mahbubar Rahman Khan ◽  
Donald James Gomes
Keyword(s):  
Alkaline Protease ◽  
Skim Milk ◽  
Bacterial Count ◽  
Protease Production ◽  
Tannery Effluent ◽  
Leather Processing ◽  
Proteolytic Activities ◽  
Key Words Bacteria ◽  
Good Number ◽  
Number Of Bacteria

Samples collected from different stages of the tannery processing were found to be alkaline. A good number of bacteria were found to be associated with the different stages of leather processing. The aerobic heterotrophic bacterial count ranged in between 11.9 × 106 and 46.7 × 106. The highest count was observed in the soaking stage and the minimum was found with the bating stage. Among 40 isolates, 31 showed positive proteolytic activities on different protein based media. Identified organisms were Bacillus subtilis (9), B. licheniformis (6), B. alcalophilus (2), B. badius (2), B. cereus (2), B. circulans (2), B. pumilus (2), B. alvei (1), B. brevis (1), B. coagulans (1), B. megaterium (1), B. polymyxa (1) and Micrococcus varians (1). Proteolytic activity was measured as zone ratio on skim milk agar which was found to be in between 1.5 and 5.8. Higher zone ratio was observed in B. subtilis (TS/1/E), B. pumilus (TS/1/S1/A3 and TD/S2/C3), B. licheniformis (TS/1/Q) and B.  badius (TD/21-D). The alkaline protease production by the nine selected isolates ranged in between 7.1 and 119.3 U/ml. Two isolates of B. pumilus (TS/1/S1/A3 and TD/S2/C3) were found to be good alkaline protease producers (119.3 and 94.8 U/ml) among the tested organisms. Biotechnologically these two isolates or their enzymes could be utilized in the tannery industry.   Key words: Bacteria, tannery, effluent, alkaline protease   D. O. I. 10.3329/ptcb.v21i1.9563   Plant Tissue Cult. & Biotech. 21(1): 53-61, 2011 (June)

Toxin Production by Psychrotrophic Bacillus spp. Present in Milk

1990 ◽  
Vol 53 (9) ◽  
pp. 790-792 ◽  
Author(s):  
M. W. GRIFFITHS
Keyword(s):  
Bacillus Cereus ◽  
Brain Heart Infusion ◽  
Bacterial Count ◽  
Toxin Production ◽  
Latex Agglutination ◽  
Brain Heart Infusion Broth ◽  
Agglutination Assay ◽  
Bacillus Spp ◽  
Latex Agglutination Assay

Using a reversed passive latex agglutination assay, about 85% of psychrotrophic Bacillus spp. tested were shown to produce diarrhoegenic toxin during growth on brain heart infusion broth at 25°C. The majority of these strains were identified as Bacillus cereus or cereus-related strains. However, a number of other species was capable of synthesizing the toxin. Further investigation of four psychrotrophic Bacilli showed that the toxin was produced during growth in milk at temperatures ranging from 6 to 21°C. Toxin production increased with increasing temperatures and was not synthesized in appreciable quantities until the bacterial count exceeded 1 × 107 cfu/ml.

scholarly journals Antibiogram of Bacteria Isolated from Tympanotonus fuscatus Var. Radula (Prosobranchia:Potamididae) Sold in Markets in Nasarawa State, Nigeria

2020 ◽  
pp. 1-9
Author(s):  
U. K. Asemota ◽  
M. D. Makut ◽  
S. O. Obiekezie ◽  
J. E. Owuna ◽  
M. O. Adamu
Keyword(s):  
Escherichia Coli ◽  
Bacterial Load ◽  
Bacterial Count ◽  
Total Bacterial Count ◽  
Bacterial Isolates ◽  
Salmonella Spp ◽  
Pseudomonas Spp ◽  
Gram Positive Bacteria ◽  
Bacillus Spp ◽  
Gram Negative Organisms

The aim of this study was to determine the antibiogram of bacterial isolates from Tympanotonus fuscatus var. radula sold in markets in Nasarawa State. Nigeria. Samples of Tympanotonus fuscatus var. radula (periwinkles) were bought from soup ingredient sellers at different sale locations in Keffi, Masaka and Orange markets and were analyzed using standard bacteriological methods. The bacterial isolates were identified using morphological, cultural and biochemical techniques. The total bacteria count varied from 1.18–3.20 x 108 CFU/g for the raw samples while the total bacterial count for the boiled samples varied from 0–1.57 x 108 CFU/g. Periwinkle samples with shells from Masaka market had the highest bacterial load with a mean total bacterial count of 2.94 x 10⁸ CFU/g and mean total coliform count of 2.80 x 10⁶ CFU/g. Raw periwinkle samples with shells had a higher bacterial load than samples without shells. There was also a drastic reduction in the bacterial load in the periwinkle samples after boiling under laboratory conditions. The bacteria isolated were Bacillus spp. and Staphylococcus aureus were the Gram-positive bacteria isolated. Enterobacter spp., Escherichia coli, Salmonella spp., Pseudomonas spp., Serratia spp. and Proteus spp. The most frequently occurring gram positive bacteria was Escherichia coli with an isolation frequency of 6(24%), the least frequently occurring was Bacillus spp., 8(32)%. Antibiotic susceptibility test showed that all the gram negative organisms exhibited sensitivity to ciprofloxacin: Escherichia coli (32 mm), Enterobacter spp. (41.5 mm), Proteus spp. (40.0 mm), Salmonella spp. (37.0 mm), Serratia spp. (26.0 mm), Pseudomonas spp. (23.0 mm). All the gram negative organisms showed marked resistance to vancomycin: Escherichia coli (12.0 mm), Enterobacter spp. (10.0 mm), Proteus spp. (11.0 mm), Salmonella spp. (5.0 mm), Serratia spp. (10.0 mm) and Pseudomonas spp. (4.5 mm).

scholarly journals Antagonistic potential of Bacillus spp. isolates against bacterial pathogens of tomato and fungal pathogen of pepper

2018 ◽  
Vol 33 (1) ◽  
pp. 9-18 ◽  
Author(s):  
Svetlana Milijasevic-Marcic ◽  
Vesna Todorovic ◽  
Olja Stanojevic ◽  
Tanja Beric ◽  
Slavisa Stankovic ◽  
...  
Keyword(s):  
Growth Inhibition ◽  
Inhibition Zone ◽  
Inhibitory Effect ◽  
Antagonistic Effect ◽  
Subtilis Strain ◽  
Vegetable Crops ◽  
Clavibacter Michiganensis Subsp ◽  
Bacillus Spp ◽  
Antagonistic Potential

In vitro antagonistic potential of eleven isolates of Bacillus spp. against two phytopathogenic bacteria and one fungus was tested in order to identify potential biocontrol agents in vegetable crops. The Bacillus spp. isolates demonstrated different levels of antagonistic effect against the tested pathogenic microorganisms. Data in the study proved Xanthomonas vesicatoria to be more sensitive to Bacillus spp. strains than Clavibacter michiganensis subsp. michiganensis. Ten B. subtilis strains induced growth inhibition of X. vesicatoria, while a strain of B. pumilus did not affect the growth of that bacterium. The largest inhibition zones against X. vesicatoria were induced by strains B-319, B-325 and B-358. The pathogenic strain C. michiganensis subsp. michiganensis was most inhibited by two B. subtilis strains (B-338 and B-348) with mean inhibition zone diameters of up to 20 mm. B. subtilis strain B-319 which was the best in inhibiting X. vesicatoria, showed the lowest inhibitory effect on C. michiganensis subsp. michiganensis. The largest growth inhibition percentage of Verticillium sp. (PGI approximately 70%) was induced by B. subtilis strains B-310 and B-322. The other B. subtilis strains showed PGI values ranging from 45% to 68%, while B. pumilus strain B-335 had the least antagonistic potential (PGI =34.43%) against the pathogen. This study identified at least one suitable biocontrol candidate, B. subtilis strain B-358, as effective in vitro against all three vegetable pathogens.

scholarly journals Effect of fortification of skim milk with coconut milk on the proximate composition and manufacture of dahi, a traditional sweet curd

2016 ◽  
Vol 2 (2) ◽  
pp. 247-252
Author(s):  
Md Rezaul Hai Rakib ◽  
Morsheda Yesmin ◽  
Md Abu Hemayet ◽  
Md Ahsanul Kabir ◽  
Md Nurul Islam
Keyword(s):  
Nutritional Value ◽  
Skim Milk ◽  
Bacterial Count ◽  
Coconut Milk ◽  
Partial Replacement ◽  
Total Bacterial Count ◽  
Microbiological Analysis ◽  
Alternative Source ◽  
Organoleptic Score ◽  
Different Levels

The experiment was conducted to measure the feasibility of partial replacement of skim milk with different levels of coconut milk in the manufacture of dahi. Skimmed milk was replaced by 0, 5, 10, 15 and 20% of coconut milk to produce Dahi designated as A, B, C, D and E. All the samples were analyzed for organoleptic, chemical and microbiological qualities. Replacement of skim milk up to 10% with coconut milk increased total organoleptic score but score decreased when level of coconut milk was 15% and 20%. Dahi manufactured by incorporating 5% coconut milk gave superior results for body and consistency and also for colour and texture. In all levels of replacement increased fat, carbohydrates, ash and total solids were obtained, while protein and pH content were significantly decreased in Dahi samples. Total bacterial count was higher in the dahi manufactured by replacement of coconut milk than control. It could be concluded that 5 and 10% replacement of skim milk with coconut milk for the manufacturing of Dahi was acceptable and reduced the production cost and they were better in compare to others, according to organoleptic, chemical and microbiological analysis. The work showed the potential of coconut as an alternative source of skim milk in dahi manufacturing with improved nutritional value and consumer acceptability.Asian J. Med. Biol. Res. June 2016, 2(2): 247-252

An Assessment of Pasteurization Treatment of Water, Media, and Milk with Respect to Bacillus Spores†

2005 ◽  
Vol 68 (4) ◽  
pp. 751-757 ◽  
Author(s):  
JOHN S. NOVAK ◽  
JEFFREY CALL ◽  
PEGGY TOMASULA ◽  
JOHN B. LUCHANSKY
Keyword(s):  
Thermal Resistance ◽  
Skim Milk ◽  
Brain Heart Infusion ◽  
Brain Heart Infusion Broth ◽  
Complete Inactivation ◽  
Milk Pasteurization ◽  
Water Media ◽  
Bacillus Spp ◽  
Bacillus Spores ◽  
Synthetic Media

This study evaluated the ability of spore-forming Bacillus spp. to resist milk pasteurization conditions from 72 to 150°C. Spores from the avirulent surrogate Sterne strain of Bacillus anthracis, as well as a representative strain of a common milk contaminant that is also a pathogen, Bacillus cereus ATCC 9818, were heated at test temperatures for up to 90 min in dH2O, brain heart infusion broth, or skim milk. In skim milk, characteristic log reductions (log CFU per milliliter) for B. anthracis spores were 0.45 after 90 min at 72°C, 0.39 after 90 min at 78°C, 8.10 after 60 min at 100°C, 7.74 after 2 min at 130°C, and 7.43 after 0.5 min at 150°C. Likewise, log reductions (log CFU per milliliter) for viable spores of B. cereus ATCC 9818 in skim milk were 0.39 after 90 min at 72°C, 0.21 after 60 min at 78°C, 7.62 after 60 min at 100°C, 7.37 after 2 min at 130°C, and 7.53 after 0.5 min at 150°C. No significant differences (P < 0.05) in thermal resistance were observed for comparisons of spores heated in dH2O or brain heart infusion broth compared with results observed in skim milk for either strain tested. However, spores from both strains were highly resistant (P < 0.05) to the pasteurization temperatures tested. As such, pasteurization alone would not ensure complete inactivation of these spore-forming pathogens in dH2O, synthetic media, or skim milk.

scholarly journals Homofermentative lactic acid bacteria of a traditional cheese, Comlek peyniri from Cappadocia region

2005 ◽  
Vol 72 (1) ◽  
pp. 19-24 ◽  
Author(s):  
Cisem Bulut ◽  
Hatice Gunes ◽  
Burcu Okuklu ◽  
Sebnem Harsa ◽  
Sevda Kilic ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Molecular Identification ◽  
Lactic Acid Production ◽  
Skim Milk ◽  
Gas Production ◽  
Central Anatolia ◽  
Rrna Gene ◽  
Ph Change ◽  
Gram Staining

Comlek peyniri is a typical artisanal cheese in Central Anatolia. This type of cheese was made by using the indigenous lactic acid bacteria (LAB) flora of cow or ewes' milk. Majority of the samples were taken from fresh cheese because the aim was to isolate homofermentative LAB. Initially 661 microbial isolates were obtained from 17 cheese samples. Only 107 were found to be homofermentative LAB. These isolates were selected and identified by using both phenotypic and molecular methods. Phenotypic identification included curd formation from skim milk, catalase test, Gram staining and light microscopy, growth at different temperatures and salt concentrations, arginine hydrolysis, gas production from glucose, and carbohydrate fermentation. Molecular identification was based on the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the 16S rRNA gene-ITS (internally transcribed spacer) region. By combining the phenotypic and molecular identification results, isolates belonging to each of the following genera were determined at species or subspecies level: 54 Lactococcus lactis subsp. lactis, 21 Enterococcus faecium, 3 Ec. faecalis, 2 Ec. durans, 10 Ec. sp., 15 Lactobacillus paracasei subsp. paracasei, and 2 Lb. casei strains. Technological characterisation was also performed by culturing each of the strains in UHT skim milk, and by monitoring pH change and lactic acid production at certain time intervals through the 24 h incubation. Results of the technological characterisation indicated that 33% of the isolates (35 strains) were capable of lowering the pH of UHT milk below 5·3 after 6 h incubation at 30 °C. Thirty four of these strains were Lc. lactis subsp. lactis, and only one was an Ec. faecium strain.

scholarly journals Isolation, characterization and molecular identification of protease producing bacteria from Tashkooh mountain located in Ahvaz, Iran

2015 ◽  
Vol 9 (2) ◽  
pp. 39-42 ◽  
Author(s):  
Arezoo Ebrahimpour ◽  
Ashraf Kariminik
Keyword(s):  
Proteolytic Activity ◽  
Thermophilic Bacteria ◽  
Skim Milk ◽  
Subtilis Strain ◽  
Industrial Enzymes ◽  
Thermophilic Microorganisms ◽  
Rrna Sequencing ◽  
Extracellular Protease Activity ◽  
Phenotypic Tests ◽  
G 13

Thermophilic microorganisms have gained worldwide importance due to their remarkable potential to produce thermostable and thermoactive enzymes that have wide applications in pharmaceuticals and industries. Therefore, the isolation of thermophilic bacteria from natural sources and their identification are very important in terms of discovering new industrial enzymes. The aim of this research was therefore the isolation of protease producing thermophilic bacteria from Tashkooh or firing mount located in Ahvaz, Iran. 8 bacterial isolates were screened. These strains were examined for the existence of extracellular protease activity. All 8 isolated bacteria showed proteolytic activity on nutrient agar with skim-milk. 3 bacteria showed their optimum growth at alkaline pH and grew maximally at different temperature in the thermophilic range and had proteolytic activity at pH 11 in 70?C and pH 9 in 55?C.The best carbon source for proteolytic activity was starch. After performing some phenotypic tests determined that all the isolates were Gram positive, endospore forming rods, aerobic, capable to produce catalase, amylase and gelatinase enzymes and they were identified as Bacillus sp. The best isolated bacteria was identified molecularly with the aid of 16S rRNA sequencing and data revealed that the Bacillus subtilis strain G-13 (GenBank accession No. KJ139434.1).The investigation confirmed that the isolate to be a true thermophile and could be a source of thermostable protease which can be exploited for pharmaceutical and industrials applications.DOI: http://dx.doi.org/10.3126/ijls.v9i2.12054 International Journal of Life Sciences 9 (2) : 2015; 39-42

scholarly journals TOTAL BACTERIA AND IDENTIFICATION OF PROTEOLYTIC RUMINAL AND RETICULUM BACTERIA FROM LOCAL KACANG GOAT

2018 ◽  
Vol 12 (1) ◽  
Author(s):  
Safika Safika ◽  
Darmawi Darmawi ◽  
Fitria Sari Ramadhani ◽  
Nurhaspika Nurhaspika ◽  
Moliwati Moliwati
Keyword(s):  
Rumen Fluid ◽  
Sequence Similarity ◽  
Skim Milk ◽  
Bacterial Count ◽  
Total Bacterial Count ◽  
Plate Method ◽  
Phylogenetic Tree Analysis ◽  
Proteolytic Bacteria ◽  
A New Species ◽  
Total Bacteria

This study aimed to calculate the total bacteria and to identify the proteolytic bacteria in rumen and reticulum of local Kacang goat. The samples used were the rumen and reticulum fluids of five goats at abattoirs in Banda Aceh. Isolation of proteolytic bacteria was carried out using skim milk agar with pour plate method and incubated at 39° C for 48 hours. The bacterial colonies morphology was observed and the total bacterial count was recorded. DNA of the widest proteolytic index colony was isolated, amplified, and sequenced. The results showed that the dominant colonies morphology was white with position inside the agar. The average of total proteolytic bacteria in goat rumen fluid and reticulum fluid were 6.416x106 CFU/mL and 2,382x107 CFU/mL, respectively. Isolates with the widest proteolytic index was Ru3 (2.5 mm) in the rumen which homology and phylogenetic tree analysis of 16S rRNA showed 93% sequence similarity with Bacillus subtilis, while in reticulum was Re1 (2.0 mm) which has 92% sequence similarity to Tatumella. It is concluded that the number of proteolytic bacteria in reticulum is greater than the number of proteolytic bacteria in the rumen of local kacang goat. Homology analysis in this study proved that the Ru3 and Re1 isolate were probably either a new species or unconfirmed species.

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