Characterization of Lactic Bacteria Isolated from Raw Milk and Their Antibacterial Activity against Bacteria as the Cause of Clinical Bovine Mastitis

The objectives of this study were the selection of lactic acid bacteria (LAB) isolated from raw milk and studying their technological properties and antibacterial activities against bacteria as the cause of cattle mastitis. Biochemical and molecular identification using 16S–23S rRNA gene spacer analysis and 16S rRNA gene sequencing highlighted the presence of three species: Lactiplantibacillus plantarum, Lactococcus lactis, and Levilactobacillus brevis. The enzymatic characterization followed by the determination of technofunctional properties showed that LAB strains did not exhibit any hemolytic effect and were able to produce protease and lipase enzymes. Isolates showed very high antagonistic activity against Gram-positive and Gram-negative bacteria by producing H2O2, bacteriocin(s), and organic acid(s). APIZYM micromethod demonstrated that all selected strains are capable of producing valine arylamidase, cystine arylamidase, N-acetyl-β-glucosaminidase, and ᾳ-mannosidase. The antibiotic susceptibility assay showed that all selected strains were sensible to the majority of tested antibiotics. Based on these results, it can be concluded that the technological properties of the selected LAB allow considering their industrial use in order to formulate bioactive functional foods or drug(s).

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Diversity of Domain V of 23S rRNA Gene Sequence in Different Enterococcus Species

Journal of Clinical Microbiology ◽

10.1128/jcm.38.11.3991-3993.2000 ◽

2000 ◽

Vol 38 (11) ◽

pp. 3991-3993 ◽

Cited By ~ 9

Author(s):

Sotirios Tsiodras ◽

Howard S. Gold ◽

Eoin P. G. Coakley ◽

Christine Wennersten ◽

Robert C. Moellering ◽

...

Keyword(s):

16S Rrna Gene Sequencing ◽

23S Rrna ◽

Rrna Gene ◽

Dna Encoding ◽

Enterococcus Casseliflavus ◽

23S Rrna Gene ◽

Rrna Gene Sequencing ◽

Biochemical Testing ◽

Enterococcus Gallinarum ◽

Domain V

The highly conserved central loop of domain V of 23S RNA (nucleotides 2042 to 2628; Escherichia coli numbering) is implicated in peptidyltransferase activity and represents one of the target sites for macrolide, lincosamide, and streptogramin B antibiotics. DNA encoding domain V (590 bp) of several species ofEnterococcus was amplified by PCR. Twenty enterococcal isolates were tested, including Enterococcus faecium (six isolates), Enterococcus faecalis, Enterococcus avium, Enterococcus durans, Enterococcus gallinarum, Enterococcus casseliflavus (two isolates of each), and Enterococcus raffinosus, Enterococcus mundtii, Enterococcus malodoratus, andEnterococcus hirae (one isolate of each). For all isolates, species identification by biochemical testing was corroborated by 16S rRNA gene sequencing. The sequence of domain V of the 23S rRNA gene from E. faecium and E. faecalis differed from those of all other enterococci. The domain V sequences of E. durans and E. hirae were identical. This was also true for E. gallinarum and E. casseliflavus. E. avium differed from E. casseliflavus by 23 bases, from E. durans by 16 bases, and from E. malodoratus by 2 bases. E. avium differed fromE. raffinosus by one base. Despite the fact that domain V is considered to be highly conserved, substantial differences were identified between several enterococcal species.

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Paenibacillus lactis sp. nov., isolated from raw and heat-treated milk

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijs.0.02822-0 ◽

2004 ◽

Vol 54 (3) ◽

pp. 885-891 ◽

Cited By ~ 80

Author(s):

Patsy Scheldeman ◽

Karen Goossens ◽

Marina Rodriguez-Diaz ◽

Annelies Pil ◽

Johan Goris ◽

...

Keyword(s):

16S Rrna ◽

16S Rrna Gene ◽

Dairy Farm ◽

16S Rrna Gene Sequencing ◽

Raw Milk ◽

Rrna Gene ◽

Heat Treated ◽

Dairy Plant ◽

Rrna Gene Sequencing ◽

High Degree

Endospore-forming bacteria were recovered from individual packages from different processing lines in a dairy plant during a tenacious periodical contamination of their UHT-milk production. Two colony types were seen, one of which was identified as Bacillus sporothermodurans. Analysis of the 16S rRNA gene of the second colony type placed these isolates within the genus Paenibacillus, with Paenibacillus lautus as the closest known relative. Moreover, over 99 % similarity was observed to the 16S rDNA sequence of MB 2035, a strain isolated previously from raw milk during a survey at dairy farms for very heat-resistant spore-forming bacteria. Nine other potentially closely related strains among the dairy farm isolates were found using rep-PCR typing. The taxonomic positions of these 19 isolates were further investigated using 16S rRNA gene sequencing and DNA–DNA hybridizations of representative strains. All 19 isolates shared a high degree of phenotypic similarity and were easily distinguished from closely related members of the genus. Anteiso-C15 : 0, C16 : 0 and iso-C15 : 0 were among the major fatty acids and the genomic DNA G+C content was 51·6–51·7 mol%. Therefore, based on their phenotypic, phylogenetic and genomic distinctiveness, these 19 strains, isolated from both raw and heat-treated milk, are placed in the genus Paenibacillus as Paenibacillus lactis sp. nov. The type strain is MB 1871T (=LMG 21940T=DSM 15596T).

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Infective Arthritis: Bacterial 23S rRNA Gene Sequencing as a Supplementary Diagnostic Method

The Open Microbiology Journal ◽

10.2174/1874285800802010085 ◽

2008 ◽

Vol 2 (1) ◽

pp. 85-88 ◽

Cited By ~ 5

Author(s):

Claus Moser ◽

Keld Andresen ◽

Anne Kjerulf ◽

Suheil Salamon ◽

Michael Kemp ◽

...

Keyword(s):

Diagnostic Method ◽

Gene Sequencing ◽

23S Rrna ◽

Rrna Gene ◽

23S Rrna Gene ◽

Rrna Gene Sequencing ◽

Infective Arthritis

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Characterization of Biosurfactant From Pseudomonas Stutzeri SJ3 for Remediation of Crude Oil-Contaminated Soil

10.21203/rs.3.rs-497731/v1 ◽

2021 ◽

Author(s):

Sekar Harikrishnan ◽

Singaram Jayalakshmi ◽

Mohamad S. Alsalhi ◽

Alager Kartick ◽

Sandhanasamy Devanesan ◽

...

Keyword(s):

Pseudomonas Stutzeri ◽

16S Rrna Gene Sequencing ◽

Antibacterial Activities ◽

Biosurfactant Production ◽

Screening Methods ◽

Rrna Gene ◽

Bacterial Strains ◽

Emulsification Index ◽

Work Production ◽

Rrna Gene Sequencing

Abstract In the present work, production of biosurfactant was studied from the bacterial strains isolated from the soil samples collected from oil contaminated sites in Karaikal ONGC, Puducherry, India. Six morphologically different hydrocarbonoclastic bacterial strains (SJ1-SJ6) isolated on oil agar plates were further screened for biosurfactant production. Based on the screening methods results of 26 mm oil displacement zone, positive results of drop collapse test, 68.14% emulsification index (E24) and 79.2% of bacterial adherence percentage, the isolate SJ3 was selected as the most potent strain and it was identified as P. stutzeri using standard biochemical and 16S rRNA gene sequencing-based methods. Optimization of the P. stutzeri strain showed 36 h incubation, 150 rpm agitation, pH 7.5, 37oC, 1% salinity, 2% glucose as carbon source and 1% yeast extract as nitrogen source were the ideal conditions for growth and the biosurfactant production was found to be growth dependent. The crude biosurfactant showed broad range of antibacterial activity against the bacterial pathogens tested. The P. stutzeri isolated from oil spill site showed biosurfactant with antibacterial activities.

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Potentials of Indigenous Bacillus thuringiensis Isolates from the soil in controlling Fusarium wilt of Cucumber cause by Fusarium oxysporum f.sp cucumerinum

Nigerian Journal of Biotechnology ◽

10.4314/njb.v37i1.14 ◽

2020 ◽

Vol 37 (1) ◽

pp. 129-137

Author(s):

P.O. Akintokun ◽

A.O. Okuwa ◽

A.R. Oloyede ◽

S.O. Adebajo ◽

A.K. Akintokun

Keyword(s):

Bacillus Thuringiensis ◽

16S Rrna ◽

16S Rrna Gene ◽

Fusarium Wilt ◽

16S Rrna Gene Sequencing ◽

Antagonistic Activity ◽

Rrna Gene ◽

Rrna Gene Sequencing ◽

Screen House

Cucumber (Cucumis sativus L.) production is generally low in Nigeria due to continuous soil nutrient limitation and diseases. However, the persistence in the use of agrochemicals for cucumber production in Nigeria is associated with high cost and deleterious effects on man, animal and the environment. This study was conducted to investigate the potentials of indigenous Bacillus thuringiensis (Bt), a spore-forming bacterium known for its insecticidal properties in controlling Fusarium wilt of cucumber. Bacillus thuringiensis strains were isolated from soil samples collected from different farm sites in Abeokuta, Nigeria, and identified phenotypically and molecularly. The in-vitro antagonistic activity of B. thuringiensis strains on F. oxysporum f.sp. cucumerinum was evaluated by dual culture method, followed by pot experiment in the screen house. 16S rRNA gene sequencing was performed on the antagonistic B. thuringiensis to confirm Bt species. The results of the in-vitro antagonistic activity revealed that most indigenous B. thuringiensis strains showed significant growth inhibition of Fusarium oxysporium f. sp. cucumerinum. Similarly, application of B. thuringiensis A and C isolates significantly suppressed the incidence of Fusarium wilt of cucumber in the screen house when compared to the control. The 16S rRNA gene sequencing technique identified the isolates A and C as Bacillus thuringiensis strain LTS-209 and Bacillus thuringiensis strain VITSJ-01, respectively. Hence, indigenous B. thuringiensis A and C isolates should be incorporated into cucumber cultivation for controlling Fusarium wilt disease of cucumber. Keywords: Cucumber, Bacillus thuringiensis, Fusarium wilt, 16S rRNA gene

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Molecular Detection of Multidrug Resistant Staphylococcus aureus Isolated from Bovine Mastitis Milk in Bangladesh

Veterinary Sciences ◽

10.3390/vetsci7020036 ◽

2020 ◽

Vol 7 (2) ◽

pp. 36

Author(s):

Md. Salauddin ◽

Mir Rowshan Akter ◽

Md. Khaled Hossain ◽

K. H. M. Nazmul Hussain Nazir ◽

Ayman Noreddin ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Bovine Mastitis ◽

Multidrug Resistant ◽

Health Concern ◽

Diffusion Method ◽

23S Rrna ◽

Nucleotide Sequence Analysis ◽

Rrna Gene ◽

Milk Samples ◽

23S Rrna Gene

The current study was conducted to isolate and identify multidrug-resistant Staphylococcus aureus (MDR-SA) from mastitis milk samples and to determine their antimicrobial susceptibility pattern. A total of 48 bovine mastitis (BM) milk samples were collected from different parts of the Rangpur division, Bangladesh. After the collection of milk samples, mastitis was confirmed using the California mastitis test. Isolation and identification of Staphylococcus aureus were performed using conventional cultural and biochemical tests as well as using molecular methods of PCR. Nucleotide sequence analysis of the 23S rRNA gene of Staphylococcus aureus was determined. The antibiogram of the isolated bacteria was conducted using the disc diffusion method. Phylogenetic analysis of 23S rRNA was done using MEGA 7, ClustalW multiple sequence alignment, and NCBI-BLAST tools, where the sequence of the isolate showed 98% to 99% identity. Antibiogram test using 15 antimicrobial agents showed that all of the Staphylococcus aureus isolates were classified as multidrug-resistant (MDR). It was found that the isolates were resistant to tetracycline, novobiocin, methicillin, vancomycin, and cephradine, and the isolates were sensitive to ciprofloxacin, azithromycin, norfloxacin, levofloxacin, gentamicin, and amoxicillin. The detection of MDR-SA in mastitis milk is alarming and represents a great public health concern. The findings of the present study help identify Staphylococcus aureus at the molecular level using 23S rRNA gene sequencing and will help select the appropriate and effective antimicrobial agent to control BM in the northern part of Bangladesh.

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Matrine-chitosan hydrogels for treating subclinical bovine mastitis by intramammary infusion—effect on milk microbiome and metabolites

10.21203/rs.3.rs-26655/v1 ◽

2020 ◽

Author(s):

Fuxin Wu ◽

Hua Zhang ◽

Hui Niu ◽

Benhai Xiong ◽

Jinjin Tong ◽

...

Keyword(s):

Molecular Mechanisms ◽

Bovine Mastitis ◽

Principal Component ◽

16S Rrna Gene Sequencing ◽

Subclinical Mastitis ◽

Rrna Gene ◽

Anticancer Properties ◽

Milk Samples ◽

Rrna Gene Sequencing ◽

Chitosan Hydrogels

Abstract BackgroundThe metabolic processes of cows undergo significant changes during subclinical mastitis, but their molecular mechanisms have not been clearly elucidated. This study investigated the changes in milk metabolites after intramammary infusion of matrine, a plant alkaloid with anticancer properties, in the form of a chitosan hydrogel into bovine mammary glands with subclinical mastitis. Infusions were continued for seven days, and milk samples were collected on day 1 and day 7 for analysis of the microbiome by 16S rRNA gene sequencing and metabolites by liquid chromatography-mass spectrometry.ResultsMatrine-chitosan hydrogels (MCHs) significantly decreased the somatic cell count on day 7 and the Simpson index indicated that microbial diversity was significantly lower on day 7 than on day 1. On day 7, the numbers of Aerococcus, Corynebacterium_1 and Staphylococcus were significantly lower, while the abundance of Firmicutes was very significantly decreased. The numbers of Probacteria increased, however. In milk samples, we identified 74 differentially expressed metabolites and the MCH infusion group had the most significantly upregulated metabolites including sphingolipids, glycerophospholipids, flavonoids and fatty acyls. Principal component analysis and the orthogonal partial least squares discriminant test confirmed good separation of the milk metabolites. The identification of active milk metabolic pathways after MCH treatment supported the known antimicrobial and anti-inflammatory properties of matrine that are associated with glycerophospholipid metabolism and the sphingolipid metabolic signaling pathways.ConciusionsThese insights into the mechanisms and the corresponding biological responses to matrine demonstrate its potential immunoregulatory activity and emphasize the need for continued investigation.

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Bacterial isolates from bryozoan Pleurocodonellina sp.: Diversity and antimicrobial potential against pathogenic bacteria

Biodiversitas Journal of Biological Diversity ◽

10.13057/biodiv/d200914 ◽

2019 ◽

Vol 20 (9) ◽

Author(s):

Meezan Ardhanu Asagabaldan ◽

Gilles Bedoux ◽

Nathalie Bourgougnon ◽

Rhesi Kristiana ◽

Diah Ayuningrum ◽

...

Keyword(s):

Pathogenic Bacteria ◽

16S Rrna Gene Sequencing ◽

Antibacterial Activities ◽

Multidrug Resistant ◽

Rrna Gene ◽

Bacterial Isolates ◽

Bacterial Strains ◽

Associated Bacteria ◽

Antimicrobial Potential ◽

Rrna Gene Sequencing

Abstract. Asagabaldan MA, Bedoux G, Bourgougnon N, Kristiana R, Ayuningrum D, Sabdono A, Trianto A, Radjasa OK. 2019. Bacterial isolates from bryozoan Pleurocodonellina sp.: Diversity and antimicrobial potential against pathogenic bacteria. Biodiversitas 20: 2528-2535. There is an urgent need to discover new compounds with antibacterial activity, which can be developed into lead structures for the treatment of human disease caused by multidrug-resistant (MDR) bacteria. In this study, we focussed on bryozoan-associated bacteria to screen them toward antibacterial activities, since the microbiome of these organisms can still be regarded as under-investigated. Most of the few publications about bryozoan-associated bacteria focused on taxonomy and the potential as producers of antibacterial natural products were neglected. Four specimens of bryozoan Pleurocodonellina sp. were collected from Teluk Awur, Jepara in Java Sea, Indonesia. Therefrom, 56 bacterial strains were isolated, and 17 displayed antibacterial activities against MDR bacteria Pseudomonas aruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Enterobacter cloacae, and methicillin-resistant Staphylococcus aureus (MRSA). Taxonomic identification of the bacteria by 16S rRNA gene sequencing revealed them belonging to the genera Virgibacillus, Pseudoalteromonas, Halomonas, and Bacillus. Most interestingly, the genus Virgibacillus was dominantly obtained from the Pleurocodonellina sp. specimens, i.e., 12 active isolates. Nevertheless, the best activities against MDR bacteria (both Gram-positive and Gram-negative) were contributed to isolates showing >99% identity to Pseudoalteromonas. The results further suggest adding the genus Virgibacillus as bacteria associated with bryozoan, since to the best of our knowledge there were no reports of this genus isolated from bryozoan.

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Discrimination of major and minor streptococci incriminated in bovine mastitis by MALDI-TOF MS fingerprinting and 16S rRNA gene sequencing

Research in Veterinary Science ◽

10.1016/j.rvsc.2020.07.027 ◽

2020 ◽

Vol 132 ◽

pp. 426-438 ◽

Cited By ~ 2

Author(s):

Mohamed E.A. Alnakip ◽

Nasreddin R. Rhouma ◽

Eman N. Abd-Elfatah ◽

Marcos Quintela-Baluja ◽

Karola Böhme ◽

...

Keyword(s):

16S Rrna ◽

16S Rrna Gene ◽

Bovine Mastitis ◽

Gene Sequencing ◽

16S Rrna Gene Sequencing ◽

Rrna Gene ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Rrna Gene Sequencing ◽

Tof Ms

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Investigation of Linezolid Resistance in Staphylococci and Enterococci

Journal of Clinical Microbiology ◽

10.1128/jcm.01929-15 ◽

2016 ◽

Vol 54 (5) ◽

pp. 1289-1294 ◽

Cited By ~ 8

Author(s):

Christopher D. Doern ◽

Jason Y. Park ◽

Michael Gallegos ◽

Debbie Alspaugh ◽

Carey-Ann D. Burnham

Keyword(s):

Susceptibility Testing ◽

Gene Sequencing ◽

Disk Diffusion ◽

23S Rrna ◽

Poor Correlation ◽

Rrna Gene ◽

Broth Microdilution ◽

Linezolid Resistance ◽

23S Rrna Gene ◽

Rrna Gene Sequencing

The objective of this study was to investigate an apparent increase in linezolid-nonsusceptible staphylococci and enterococci following a laboratory change in antimicrobial susceptibility testing from disk diffusion to an automated susceptibility testing system. Isolates with nonsusceptible results (n= 27) from Vitek2 were subjected to a battery of confirmatory testing which included disk diffusion, Microscan broth microdilution, Clinical and Laboratory Standards Institute (CLSI) reference broth microdilution, gradient diffusion (Etest), 23S rRNA gene sequencing, andcfrPCR. Our results show that there is poor correlation between methods and that only 70 to 75% of isolates were confirmed as linezolid resistant with alternative phenotypic testing methods (disk diffusion, Microscan broth microdilution, CLSI broth microdilution, and Etest). 23S rRNA gene sequencing identified mutations previously associated with linezolid resistance in 16 (59.3%) isolates, and thecfrgene was detected in 3 (11.1%) isolates. Mutations located at positions 2576 and 2534 of the 23S rRNA gene were most common. In addition, two previously undescribed variants (at positions 2083 and 2345 of the 23S rRNA gene) were also identified and may contribute to linezolid resistance.

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