scholarly journals Isolation from Sewage Influent and Characterization of Novel Staphylococcus aureus Bacteriophages with Wide Host Ranges and Potent Lytic Capabilities

2009 ◽  
Vol 75 (13) ◽  
pp. 4483-4490 ◽  
Author(s):  
Aidan J. Synnott ◽  
Ying Kuang ◽  
Miki Kurimoto ◽  
Keiko Yamamichi ◽  
Hidetomo Iwano ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Host Range ◽  
Protein A ◽  
Bovine Mastitis ◽  
Economic Consequences ◽  
Resistant Bacteria ◽  
Bacteriophage Therapy ◽  
Transmission Electron ◽  
65 H

ABSTRACT Staphylococcus aureus is a gram-positive pathogen that causes a variety of diseases, including bovine mastitis, which has severe economic consequences. Standard antibiotic treatment results in selection of resistant strains, leading to a need for alternative treatments, such as bacteriophage therapy. Forty-nine S. aureus isolates were obtained from the milk of mastitic cows for use in screening of staphylococcal phages. Fifteen isolates which were positive for both coagulase and hemolysin were assayed by PCR for variation in the X region and the immunoglobulin G-binding region of the protein A gene (spa) and in the carboxy terminus of the coagulase gene (coa) and for the presence of enterotoxin C, G, H, and I genes. The host ranges of 52 phages isolated from sewage influent were determined by performing spot tests with the 15 S. aureus isolates, and two phages were subsequently chosen for further analysis. ΦSA039 had the widest host range, producing clear plaques on 13 of the 15 isolates (87%), while ΦSA012 produced clear plaques on 8 isolates (53%) and was the only phage that produced a clear plaque on a nonmastitic S. aureus strain. Transmission electron microscopy revealed that the phages were similar sizes and belonged to the Myoviridae family. Measurement of optical densities during coculture with S. aureus isolates confirmed the breadth of the ΦSA039 host range and showed that ΦSA012 had potent lytic capability. ΦSA012-resistant bacteria did not appear for three of seven isolates tested (43%) after 65 h of incubation. These two phages are proposed as candidates for phage therapy of bovine mastitis.

scholarly journals Proteomic Characterization of Lytic Bacteriophages of Staphylococcus aureus Isolated from Sewage Affluent of India

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Kamalpreet Kaur Sangha ◽  
B. V. Sunil Kumar ◽  
Ravi Kant Agrawal ◽  
Dipak Deka ◽  
Ramneek Verma
Keyword(s):  
Staphylococcus Aureus ◽  
Bovine Mastitis ◽  
Soft Agar ◽  
Culture Collection ◽  
Economic Consequences ◽  
Icosahedral Symmetry ◽  
Bacteriophage Therapy ◽  
Tryptone Soya Agar ◽  
Isolation And Characterization

Staphylococcus aureus is a Gram-positive bacterium that causes a variety of diseases, including bovine mastitis, which has severe economic consequences. Standard antibiotic treatment results in selection of resistant strains, leading to need for an alternative treatment such as bacteriophage therapy. Present study describes isolation and characterization of a staphylococcal phage from sewage samples. S. aureus isolates obtained from microbial type culture collection (MTCC), Chandigarh, India, were used to screen staphylococcal phages. A phage designated as ΦMSP was isolated from sewage samples by soft agar overlay method. It produced clear plaques on tryptone soya agar overlaid with S. aureus. Transmission electron microscopy revealed that the phage had an icosahedral symmetry. It had 5 major proteins and possessed a peptidoglycan hydrolase corresponding to 70 kDa. ΦMSP infection induced 26 proteins to be uniquely expressed in S. aureus. This phage can be proposed as a candidate phage to treat staphylococcal infections.

scholarly journals JD419, a Staphylococcus aureus Phage With a Unique Morphology and Broad Host Range

2021 ◽  
Vol 12 ◽  
Author(s):  
Tingting Feng ◽  
Sebastian Leptihn ◽  
Ke Dong ◽  
Belinda Loh ◽  
Yan Zhang ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Host Range ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
Complete Characterization ◽  
Resistant Bacteria ◽  
Broad Host Range ◽  
Gene Engineering ◽  
Mrsa Strains

Phage therapy represents a possible treatment option to cure infections caused by multidrug-resistant bacteria, including methicillin and vancomycin-resistant Staphylococcus aureus, to which most antibiotics have become ineffective. In the present study, we report the isolation and complete characterization of a novel phage named JD219 exhibiting a broad host range able to infect 61 of 138 clinical strains of S. aureus tested, which included MRSA strains as well. The phage JD419 exhibits a unique morphology with an elongated capsid and a flexible tail. To evaluate the potential of JD419 to be used as a therapeutic phage, we tested the ability of the phage particles to remain infectious after treatment exceeding physiological pH or temperature. The activity was retained at pH values of 6.0–8.0 and below 50°C. As phages can contain virulence genes, JD419’s complete genome was sequenced. The 45509 bp genome is predicted to contain 65 ORFs, none of which show homology to any known virulence or antibiotic resistance genes. Genome analysis indicates that JD419 is a temperate phage, despite observing rapid replication and lysis of host strains. Following the recent advances in synthetic biology, JD419 can be modified by gene engineering to remove prophage-related genes, preventing potential lysogeny, in order to be deployed as a therapeutic phage.

Comparative sequence analysis of spa gene of Staphylococcus aureus isolated from bovine mastitis: characterization of an unusual spa gene variant

2006 ◽  
Vol 73 (3) ◽  
pp. 322-327 ◽  
Author(s):  
Amr El-Sayed ◽  
Jörg Alber ◽  
Christoph Lämmler ◽  
Amir Abdulmawjood ◽  
Michael Zschöck ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Protein A ◽  
Bovine Mastitis ◽  
Clinical Mastitis ◽  
Comparative Sequence Analysis ◽  
Border Region ◽  
Binding Domains ◽  
Igg Binding ◽  
Spa Gene

The protein A encoding gene spa of four Staphylococcus aureus strains isolated from bovine clinical mastitis was amplified by PCR and sequenced. The four strains were selected after an initial screening of spa gene of 41 strains isolated from mastitic cows and were subjected to detailed investigations. According to the sequencing results the spa gene of three strains (M1, M2, M3) appeared with gene segments encoding five (E, D, A, B and C) and four (E, A, B and C) IgG binding domains for two (M1, M3) and one (M2) strain, respectively and with gene segments encoding four, two and two repeats of the octapeptide Xr-repeats for the strains M1, M2 and M3, respectively. For the remaining Staph. aureus strain (M4) gene segments encoding IgG binding domains E, D and A and a new domain BC with a size of 219 bp could be observed. The BC domain appears, with a deletion of a 123 bp segment from the border region between both domains, as fused domain of both previously characterized domains. The Xr-region of this strain had 11 octapeptide repeats.

scholarly journals Characterization of Passage-Selected Vancomycin-Resistant Staphylococcus aureus Strains of Diverse Parental Backgrounds

2000 ◽  
Vol 44 (2) ◽  
pp. 294-303 ◽  
Author(s):  
Richard F. Pfeltz ◽  
Vineet K. Singh ◽  
Jennifer L. Schmidt ◽  
Michael A. Batten ◽  
Christopher S. Baranyk ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Walls ◽  
Methicillin Resistance ◽  
Surface Hydrophobicity ◽  
Whole Cell ◽  
Transmission Electron ◽  
Vancomycin Mics ◽  
Vancomycin Resistant ◽  
Doubling Times

ABSTRACT A series of 12 Staphylococcus aureus strains of various genetic backgrounds, methicillin resistance levels, and autolytic activities were subjected to selection for the glycopeptide-intermediate S. aureus (GISA) susceptibility phenotype on increasing concentrations of vancomycin. Six strains acquired the phenotype rapidly, two did so slowly, and four failed to do so. The vancomycin MICs for the GISA strains ranged from 4 to 16 μg/ml, were stable to 20 nonselective passages, and expressed resistance homogeneously. Neither ease of acquisition of the GISA phenotype nor the MIC attained correlated with methicillin resistance hetero- versus homogeneity or autolytic deficiency or sufficiency. Oxacillin MICs were generally unchanged between parent and GISA strains, although the mec members of both isogenic methicillin-susceptible and methicillin-resistant pairs acquired the GISA phenotype more rapidly and to higher MICs than did their susceptible counterparts. Transmission electron microscopy revealed that the GISA strains appeared normal in the absence of vancomycin but had thickened and diffuse cell walls when grown with vancomycin at one-half the MIC. Common features among GISAs were reduced doubling times, decreased lysostaphin susceptibilities, and reduced whole-cell and zymographic autolytic activities in the absence of vancomycin. This, with surface hydrophobicity differences, indicated that even in the absence of vancomycin the GISA cell walls differed from those of the parents. Autolytic activities were further reduced by the inclusion of vancomycin in whole-cell and zymographic studies. The six least vancomycin-susceptible GISA strains exhibited an increased capacity to remove vancomycin from the medium versus their parent lines. This study suggests that while some elements of the GISA phenotype are strain specific, many are common to the phenotype although their expression is influenced by genetic background. GISA strains with similar glycopeptide MICs may express individual components of the phenotype to different extents.

scholarly journals Whole-Genome Epidemiology and Characterization of Methicillin-Susceptible Staphylococcus aureus ST398 From Retail Pork and Bulk Tank Milk in Shandong, China

2021 ◽  
Vol 12 ◽  
Author(s):  
Xiaonan Zhao ◽  
Ming Hu ◽  
Cui Zhao ◽  
Qing Zhang ◽  
Lulu Li ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Protein A ◽  
Whole Genome ◽  
Staphylococcal Protein A ◽  
Sequencing Analysis ◽  
Bulk Tank Milk ◽  
Predominant Type ◽  
Bulk Tank ◽  
The One

Staphylococcus aureus (S. aureus) is now regarded as a zoonotic agent. Methicillin-susceptible S. aureus (MSSA) ST398 is a livestock-associated bacterium that is most prevalent in China, but there are currently no data available for Shandong. Therefore, the aim of this study was to investigate the epidemiology and characterization of MSSA ST398 from retail pork and bulk tank milk (BTM) in Shandong. A total of 67 S. aureus isolates were collected from retail pork between November 2017 and June 2018. Among the isolates, high antimicrobial resistance rates were observed for penicillin (97.0%), and 92.5% of the isolates were multi-drug resistant (MDR). Eight sequence types (STs) were identified in the retail pork isolates, and the predominant type was ST15 (n=26), which was followed by ST398 (n=14). Staphylococcal protein A gene (spa) typing identified spa types t034 and t1255 in MSSA ST398 from retail pork. Using whole-genome sequencing analysis, we described the phylogeny of 29 MSSA ST398 isolates that were obtained from retail pork (n=14) and BTM (n=15). The phylogenetic tree showed that the MSSA ST398 isolates from different sources had the same lineage. Among the 29 MSSA ST398 isolates, five resistance genes were detected, and all isolates carried DHA-1. Fifteen toxin genes were detected, and all isolates carried eta, hla, and hlb. In conclusion, this study found that a high risk for MSSA ST398 was present in retail pork and BTM. These findings have major implications for how investigations of MSSA ST398 outbreaks should be conducted in the One-Health context.

Isolation and characterization of two new Staphylococcus aureus bacteriophages with potential to infect distinct bacteria from bovine mastitis

2021 ◽  
Author(s):  
Bibiana Martins Barasuol ◽  
Juliana Felipetto Cargnelutti ◽  
Luis Antônio Sangioni ◽  
Daniela Isabel Brayer Pereira ◽  
Ana Paula Muterle Varela ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Bovine Milk ◽  
Bovine Mastitis ◽  
Gc Content ◽  
Etiologic Agent ◽  
Cure Rate ◽  
Long Tail ◽  
Isolation And Characterization ◽  
Double Stranded Dna

Abstract Bovine mastitis is an important disease of dairy cows, and Staphylococcus aureus is the etiologic agent most prevalent among the microorganisms. Mastitis caused by S. aureus present low cure rate with antimicrobials treatment and low vaccines efficacy. Bacteriophages or phages have been considered as an alternative for treating this disease. This study, we isolated and characterized two new S. aureus phages, namely B_UFSM4 and B_UFSM5, from bovine milk of cows with mastitis. The adsorptions rates were 10–20 min for B_UFSM4 and 20–30 min for B_UFSM5. Phages activities were relatively stable at pH 3–11; however, at temperatures of 50 °C-60ºC-70ºC/60 min, the phages were completely inactivated. These viruses presented infectivity in various bacteria isolated from bovine mastitis, where the lytic activity of phages B_UFSM4 and B_UFSM5 were 34.2%(13/38) and 42.1%(16/38), respectively, including isolates from S. aureus, Pseudomonas aeruginosa, Staphylococcus sciuri, and Rothia terrae. The complete genomes of B_UFSM4 and B_UFSM5 have 41.396 bp and 41.829 bp, with GC-content 33.97% and 33.98%, respectively. Both phages comprise 61 putative ORFs. The viruses have double stranded DNA and linear architecture. Phylogenic similarity was observed by proteome with Staphylococcus prophage phiPV83 (45,536 nt), Staphylococcus phage CN125 (44,492 nt) and Staphylococcus phage JS01 (43,458 nt). Based on the morphology, the phages belong to Siphoviridae family, presenting icosahedral head with a long tail, Caudovirales order and Biseptimavirus genus. Thus, two S. aureus phages (B_UFSM4 and B_UFSM5) were isolated and characterized, and these phages can be used as therapeutic or prophylactic candidates against S. aureus infections in cattle mastitis.

scholarly journals A Novel Genotype and First Record of Trypanosoma lainsoni in Argentina

Pathogens ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 731
Author(s):  
Anahí G. Díaz ◽  
Paula G. Ragone ◽  
Fanny Rusman ◽  
Noelia Floridia-Yapur ◽  
Rubén M. Barquez ◽  
...  
Keyword(s):  
Host Range ◽  
First Record ◽  
Morphological Characterization ◽  
Pairwise Identity ◽  
Wild Mammals ◽  
Transmission Electron ◽  
Chaco Region ◽  
Veterinary Importance ◽  
First Time

Trypanosomes are a group of parasitic flagellates with medical and veterinary importance. Despite many species having been described in this genus, little is known about many of them. Here, we report a genetic and morphological characterization of trypanosomatids isolated from wild mammals from the Argentine Chaco region. Parasites were morphologically and ultrastructurally characterized by light microscopy and transmission electron microscopy. Additionally, 18s rRNA and gGAPDH genes were sequenced and analyzed using maximum likelihood and Bayesian inference. Morphological characterization showed clear characteristics associated with the Trypanosoma genus. The genetic characterization demonstrates that the studied isolates have identical sequences and a pairwise identity of 99% with Trypanosoma lainsoni, which belongs to the clade of lizards and snakes/rodents and marsupials. To date, this species had only been found in the Amazon region. Our finding represents the second report of T. lainsoni and the first record for the Chaco region. Furthermore, we ultrastructurally described for the first time the species. Finally, the host range of T. lainsoni was expanded (Leopardus geoffroyi, Carenivora, Felidae; and Calomys sp., Rodentia, Cricetidae), showing a wide host range for this species.

scholarly journals Characterization of two newly isolated Staphylococcus aureus bacteriophages from Japan belonging to the genus Silviavirus

2020 ◽  
Vol 165 (10) ◽  
pp. 2355-2359
Author(s):  
Naoya Kitamura ◽  
Eri Sasabe ◽  
Shigenobu Matsuzaki ◽  
Masanori Daibata ◽  
Tetsuya Yamamoto
Keyword(s):  
Staphylococcus Aureus ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Dna Sequences ◽  
Related Gene ◽  
Bacteriophage Therapy ◽  
Dna Rearrangements ◽  
Site Specific

Abstract Two Staphylococcus aureus bacteriophages, KSAP7 and KSAP11, were isolated from sewage and characterized. Based on morphology and DNA sequences, they were assigned to the genus Silviavirus, subfamily Twortvirinae, family Herelleviridae, whose members are hypothesized to be suitable for bacteriophage therapy. The KSAP7 and KSAP11 genomes were 137,950 and 138,307 bp in size, respectively. Although their DNA sequences were almost identical, evidence of site-specific DNA rearrangements was found in two regions. Changes in the number of PIEPEK amino acid sequence repeats encoded by orf10 and the insertion/deletion of a 541-bp sequence that includes a possible tail-related gene were identified.

scholarly journals Efficacy of Bacteriophages Against Staphylococcus aureus Isolates from Bovine Mastitis

2020 ◽  
Vol 13 (3) ◽  
pp. 35 ◽  
Author(s):  
Isabel Titze ◽  
Tatiana Lehnherr ◽  
Hansjörg Lehnherr ◽  
Volker Krömker
Keyword(s):  
Staphylococcus Aureus ◽  
Host Range ◽  
Bacterial Infections ◽  
Plaque Assay ◽  
Bovine Milk ◽  
Bovine Mastitis ◽  
Raw Milk ◽  
Pasteurized Milk

The lytic efficacy of bacteriophages against Staphylococcus aureus isolates from bovine milk was investigated in vitro, regarding possible applications in the therapy of udder inflammation caused by bacterial infections (mastitis). The host range of sequenced, lytic bacteriophages was determined against a collection of 92 Staphylococcus (S.) aureus isolates. The isolates originated from quarter foremilk samples of clinical and subclinical mastitis cases. A spot test and a subsequent plaque assay were used to determine the phage host range. According to their host range, propagation and storage properties, three phages, STA1.ST29, EB1.ST11, and EB1.ST27, were selected for preparing a bacteriophage mixture (1:1:1), which was examined for its lytic activity against S. aureus in pasteurized and raw milk. It was found that almost two thirds of the isolates could be lysed by at least one of the tested phages. The bacteriophage mixture was able to reduce the S. aureus germ density in pasteurized milk and its reduction ability was maintained in raw milk, with only a moderate decrease compared to the results in pasteurized milk. The significant reduction ability of the phage mixture in raw milk promotes further in vivo investigation.

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