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March 28, 2022
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Article

GVPC Medium Manufactured without Oxygen Improves the Growth of Legionella spp

Publicated to: Microbiology Spectrum. 10 (2): e0240121-e0240121 - 2022-04-01 10(2), DOI: 10.1128/spectrum.02401-21

Authors:

Casino, P; López, A; Peiró, S; Ríos, M; Ríos, S; Porta, A; Agustí, G; Asensio, D; Marqués, AM; Piqué, N
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Affiliations

Department of Biochemistry and Biothecnology, Human Nutrition Unit, Universitat Rovira i Virgili, Reus, Catalonia, Spain. - Author
Department of Quality Control, Reactivos para Diagnóstico, S.L., Barcelona, Catalonia, Spain. - Author
Department of Statistics, Biology Faculty, Universitat de Barcelona, Barcelona, Catalonia, Spain. - Author
Institut de Recerca en Nutrició i Seguretat Alimentària de la UB (INSA-UB), Universitat de Barcelona, Barcelona, Catalonia, Spain. - Author
Microbiology Section, Department of Biology, Healthcare and Environment, Faculty of Pharmacy and Food Sciences, Universitat de Barcelona, Barcelona, Catalonia, Spain. - Author
Reactivos Diagnost SL, Dept Qual Control, Barcelona, Catalonia, Spain - Author
Univ Barcelona, Biol Fac, Dept Genet Microbiol & Stat, Barcelona, Catalonia, Spain - Author
Univ Barcelona, Fac Pharm & Food Sci, Dept Biol Healthcare & Environm, Microbiol Sect, Barcelona, Catalonia, Spain - Author
Univ Barcelona, Inst Recerca Nutricio & Seguretat Alimentaria UB, Barcelona, Catalonia, Spain - Author
Univ Rovira & Virgili, Dept Biochem & Biothecnol, Human Nutr Unit, Reus, Catalonia, Spain - Author
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Abstract

Glycine-vancomycin-polymyxin-cycloheximide agar (GVPC) is a recommended medium for the detection of Legionella spp. in water samples. However, its quality could be improved in terms of recovery of Legionella spp. and selectivity properties. Modifications were introduced in GVPC manufacture: autoclaving conditions (115°C, 15 min) and atmosphere during component-stirring (removal of oxygen and N2 injection). The use of softer autoclaving conditions (115°C, 15 min) improved the growth of Legionella anisa by the spiral method and Legionella pneumophila after membrane filtration. The medium manufactured with O2 removal and autoclaving for 15 min at 115°C allowed a faster growth of L. pneumophila (colonies visible at day 2) and a notable increase of L. anisa growth (colonies appearing at day 3, and statistically significant numbers of CFU at day 5). After 3 to 5 days of incubation, the improved media showed higher selectivity properties, particularly for Enterococcus faecalis ATCC 29212 and Pseudomonas aeruginosa ATCC 9027. A further improvement was achieved by the addition of N2 during ingredient stirring, leading to a statistically significant faster growth of L. pneumophila at days 2 and 3 and L. anisa at day 3. Selectivity properties were also enhanced, resulting in the complete inhibition of both E. faecalis strains and Escherichia coli and complete-partial inhibition of P. aeruginosa. Oxygen removal during GVPC manufacture using a vacuum pump system promotes the growth of L. pneumophila and L. anisa, and markedly inhibits the growth of E. coli, P. aeruginosa, and E. faecalis. IMPORTANCE Currently, GVPC is a recommended medium for the detection of Legionella spp. in water samples. However, recovery of Legionella spp. and selectivity properties can be improved. GVPC medium manufactured without oxygen improved the growth of Legionella pneumophila and Legionella anisa. Oxygen removal during GVPC manufacture also improved selectivity properties. A further improvement was achieved by the addition of N2 during ingredient stirring, leading to a faster growth of L. pneumophila at days 2 and 3 and L. anisa at day 3 and enhancement of selectivity properties. The introduction of the modified GVPC medium in routine practice can allow a better detection of Legionella spp. in water samples.
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Keywords

adsorptionanisacatalaseenterococcus faecalisgvpclegionella anisaoxygenpneumophilapseudomonas aeruginosarecoveryselectivitysuperoxidesystemCulture mediaCycloheximideEnterococcus faecalisEscherichia coliGlycineGvpcLegionellaLegionella anisaLegionella pneumophilaOutbreakOxygenPolymyxinsPseudomonas aeruginosaRecoverySelectivityVancomycinWaterWater microbiology

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal Microbiology Spectrum due to its progression and the good impact it has achieved in recent years, according to the agency Scopus (SJR), it has become a reference in its field. In the year of publication of the work, 2022, it was in position , thus managing to position itself as a Q1 (Primer Cuartil), in the category Ecology.

Independientemente del impacto esperado determinado por el canal de difusión, es importante destacar el impacto real observado de la propia aportación.

Según las diferentes agencias de indexación, el número de citas acumuladas por esta publicación hasta la fecha 2026-04-04:

  • WoS: 4
  • Scopus: 5
  • Europe PMC: 3
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Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2026-04-04:

  • The use, from an academic perspective evidenced by the Altmetric agency indicator referring to aggregations made by the personal bibliographic manager Mendeley, gives us a total of: 15.
  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 15 (PlumX).

With a more dissemination-oriented intent and targeting more general audiences, we can observe other more global scores such as:

  • The Total Score from Altmetric: 2.
  • The number of mentions on the social network X (formerly Twitter): 2 (Altmetric).

It is essential to present evidence supporting full alignment with institutional principles and guidelines on Open Science and the Conservation and Dissemination of Intellectual Heritage. A clear example of this is:

  • The work has been submitted to a journal whose editorial policy allows open Open Access publication.
  • Assignment of a Handle/URN as an identifier within the deposit in the Institutional Repository: http://hdl.handle.net/20.500.11797/imarina9247881
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Awards linked to the item

This project was possible thanks to the grant from Doctorats Industrials (2019 DI 92) from Generalitat de Catalunya.
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