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Article ; Online: A neglected and emerging antimicrobial resistance gene encodes for a serine-dependent macrolide esterase.

Dhindwal, Poonam / Thompson, Charis / Kos, Daniel / Planedin, Koa / Jain, Richa / Jelinski, Murray / Ruzzini, Antonio

Proceedings of the National Academy of Sciences of the United States of America

2023 Volume 120, Issue 8, Page(s) e2219827120

Abstract: The discovery of unreported antimicrobial resistance genes (ARGs) remains essential. Here, we report the identification and preliminary characterization of an α/β-hydrolase that inactivates macrolides. This serine-dependent macrolide esterase co-occurs ... ...

Abstract	The discovery of unreported antimicrobial resistance genes (ARGs) remains essential. Here, we report the identification and preliminary characterization of an α/β-hydrolase that inactivates macrolides. This serine-dependent macrolide esterase co-occurs with emerging ARGs in the environment, animal microbiomes, and pathogens.
MeSH term(s)	Animals ; Anti-Bacterial Agents/pharmacology ; Macrolides/pharmacology ; Drug Resistance, Bacterial/genetics ; Esterases/genetics ; Serine/genetics ; Genes, Bacterial
Chemical Substances	Anti-Bacterial Agents ; Macrolides ; Esterases (EC 3.1.-) ; Serine (452VLY9402)
Language	English
Publishing date	2023-02-15
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	209104-5
ISSN	1091-6490 ; 0027-8424
ISSN (online)	1091-6490
ISSN	0027-8424
DOI	10.1073/pnas.2219827120
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Article ; Online: Gene Flow and Molecular Innovation in Bacteria.

Ruzzini, Antonio C / Clardy, Jon

Current biology : CB

2016 Volume 26, Issue 18, Page(s) R859–R864

Abstract: The small molecules produced by environmental bacteria have been mainstays of both chemical and biological research for decades, and some have led to important therapeutic interventions. These small molecules have been shaped by natural selection as they ...

Abstract	The small molecules produced by environmental bacteria have been mainstays of both chemical and biological research for decades, and some have led to important therapeutic interventions. These small molecules have been shaped by natural selection as they evolved to fulfill changing functional roles in their native environments. This minireview describes some recent systematic studies providing illustrative examples that involve the acquisition and alteration of genetic information for molecular innovation by bacteria in well-defined environments. Two different bacterial genera are featured, Pseudonocardia and Salinispora, and, although the small-molecule repertoires of both have benefited from horizontal gene transfer, Pseudonocardia spp. have relied on plasmid-based tactics while Salinispora spp. have relied on chromosomally integrated genomic islands.
Language	English
Publishing date	2016-09-26
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	1071731-6
ISSN	1879-0445 ; 0960-9822
ISSN (online)	1879-0445
ISSN	0960-9822
DOI	10.1016/j.cub.2016.08.004
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Article ; Online: Evaluating approved and alternative treatments against an oxytetracycline-resistant bacterium responsible for European foulbrood disease in honey bees.

Masood, Fatima / Thebeau, Jenna M / Cloet, Allyssa / Kozii, Ivanna V / Zabrodski, Michael W / Biganski, Sarah / Liang, Jenny / Marta Guarna, M / Simko, Elemir / Ruzzini, Antonio / Wood, Sarah C

Scientific reports

2022 Volume 12, Issue 1, Page(s) 5906

Abstract: European foulbrood (EFB) is a disease of honey bee larvae caused by Melissococcus plutonius. In North America, oxytetracycline (OTC) is approved to combat EFB disease though tylosin (TYL) and lincomycin (LMC) are also registered for use against American ... ...

Abstract	European foulbrood (EFB) is a disease of honey bee larvae caused by Melissococcus plutonius. In North America, oxytetracycline (OTC) is approved to combat EFB disease though tylosin (TYL) and lincomycin (LMC) are also registered for use against American foulbrood disease. Herein, we report and characterize an OTC-resistant M. plutonius isolate from British Columbia, Canada, providing an antimicrobial sensitivity to the three approved antibiotics and studying their abilities to alter larval survival in an in vitro infection model. Specifically, we investigated OTC, TYL, and LMC as potential treatment options for EFB disease using laboratory-reared larvae infected with M. plutonius. The utility of the three antibiotics were compared through an experimental design that either mimicked metaphylaxis or antimicrobial intervention. At varying concentrations, all three antibiotics prevented clinical signs of EFB disease following infection with M. plutonius 2019BC1 in vitro. This included treatment with 100 μg/mL of OTC, a concentration that was ~ 3× the minimum inhibitory concentration measured to inhibit the strain in nutrient broth. Additionally, we noted high larval mortality in groups treated with doses of OTC corresponding to ~ 30× the dose required to eliminate bacterial growth in vitro. In contrast, TYL and LMC were not toxic to larvae at concentrations that exceed field use. As we continue to investigate antimicrobial resistance (AMR) profiles of M. plutonius from known EFB outbreaks, we expect a range of AMR phenotypes, reiterating the importance of expanding current therapeutic options along with alternative management practices to suppress this disease.
MeSH term(s)	Animals ; Anti-Bacterial Agents/pharmacology ; Anti-Bacterial Agents/therapeutic use ; Bacteria ; Bees ; British Columbia ; Larva ; Oxytetracycline/pharmacology ; Oxytetracycline/therapeutic use ; United States
Chemical Substances	Anti-Bacterial Agents ; Oxytetracycline (X20I9EN955)
Language	English
Publishing date	2022-04-07
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-022-09796-4
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Article ; Online: Establishment of apiary-level risk of American foulbrood through the detection of Paenibacillus larvae spores in pooled, extracted honey in Saskatchewan.

Zabrodski, Michael W / Epp, Tasha / Wilson, Geoff / Moshynskyy, Igor / Sharafi, Mohsen / Reitsma, Lara / Castano Ospina, Mateo / DeBruyne, Jessica E / Wentzell, Alexandra / Wood, Sarah C / Kozii, Ivanna V / Klein, Colby D / Thebeau, Jenna / Sobchishin, LaRhonda / Ruzzini, Antonio C / Simko, Elemir

Scientific reports

2022 Volume 12, Issue 1, Page(s) 8848

Abstract: Paenibacillus larvae, the causative agent of American foulbrood (AFB), produces spores that may be detectable within honey. We analyzed the spore content of pooled, extracted honey from 52 large-scale (L) and 64 small-scale (S) Saskatchewan beekeepers ... ...

Abstract	Paenibacillus larvae, the causative agent of American foulbrood (AFB), produces spores that may be detectable within honey. We analyzed the spore content of pooled, extracted honey from 52 large-scale (L) and 64 small-scale (S) Saskatchewan beekeepers over a two-year period (2019-2020). Our objectives were: (i) establish reliable prognostic reference ranges for spore concentrations in extracted honey to determine future AFB risk at the apiary level; (ii) identify management practices as targets for mitigation of risk. P. larvae spores were detected in 753 of 1476 samples (51%). Beekeepers were stratified into low (< 2 spores/gram), moderate (2- < 100 spores/gram), and high (≥ 100 spores/gram) risk categories. Of forty-nine L beekeepers sampled in 2019, those that reported AFB in 2020 included 0/26 low, 3/18 moderate, and 3/5 high risk. Of twenty-seven L beekeepers sampled in 2020, those that reported AFB in 2021 included 0/11 low, 2/14 moderate, and 1/2 high risk. Predictive modelling included indoor overwintering of hives, purchase of used equipment, movement of honey-producing colonies between apiaries, beekeeper demographic, and antimicrobial use as risk category predictors. Saskatchewan beekeepers with fewer than 2 spores/gram in extracted honey that avoid high risk activities may be considered at low risk of AFB the following year.
MeSH term(s)	Animals ; Bees ; Honey ; Larva ; Paenibacillus ; Paenibacillus larvae ; Saskatchewan ; Spores, Bacterial ; United States
Language	English
Publishing date	2022-05-25
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-022-12856-4
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Article ; Online: Comparison of individual hive and apiary-level sample types for spores of Paenibacillus larvae in Saskatchewan honey bee operations.

Zabrodski, Michael W / DeBruyne, Jessica E / Wilson, Geoff / Moshynskyy, Igor / Sharafi, Mohsen / Wood, Sarah C / Kozii, Ivanna V / Thebeau, Jenna / Klein, Colby D / Medici de Mattos, Igor / Sobchishin, LaRhonda / Epp, Tasha / Ruzzini, Antonio C / Simko, Elemir

PloS one

2022 Volume 17, Issue 2, Page(s) e0263602

Abstract: Three commercial honey bee operations in Saskatchewan, Canada, with outbreaks of American foulbrood (AFB) and recent or ongoing metaphylactic antibiotic use were intensively sampled to detect spores of Paenibacillus larvae during the summer of 2019. Here, ...

Abstract	Three commercial honey bee operations in Saskatchewan, Canada, with outbreaks of American foulbrood (AFB) and recent or ongoing metaphylactic antibiotic use were intensively sampled to detect spores of Paenibacillus larvae during the summer of 2019. Here, we compared spore concentrations in different sample types within individual hives, assessed the surrogacy potential of honey collected from honey supers in place of brood chamber honey or adult bees within hives, and evaluated the ability of pooled, extracted honey to predict the degree of spore contamination identified through individual hive testing. Samples of honey and bees from hives within apiaries with a recent, confirmed case of AFB in a single hive (index apiaries) and apiaries without clinical evidence of AFB (unaffected apiaries), as well as pooled, apiary-level honey samples from end-of-season extraction, were collected and cultured to detect and enumerate spores. Only a few hives were heavily contaminated by spores in any given apiary. All operations were different from one another with regard to both the overall degree of spore contamination across apiaries and the distribution of spores between index apiaries and unaffected apiaries. Within operations, individual hive spore concentrations in unaffected apiaries were significantly different from index apiaries in the brood chamber (BC) honey, honey super (HS) honey, and BC bees of one of three operations. Across all operations, BC honey was best for discriminating index apiaries from unaffected apiaries (p = 0.001), followed by HS honey (p = 0.06), and BC bees (p = 0.398). HS honey positively correlated with both BC honey (rs = 0.76, p < 0.0001) and bees (rs = 0.50, p < 0.0001) and may be useful as a surrogate for either. Spore concentrations in pooled, extracted honey seem to have predictive potential for overall spore contamination within each operation and may have prognostic value in assessing the risk of future AFB outbreaks at the apiary (or operation) level.
MeSH term(s)	Animal Diseases/diagnosis ; Animal Diseases/epidemiology ; Animal Diseases/prevention & control ; Animals ; Anti-Bacterial Agents/therapeutic use ; Beekeeping/statistics & numerical data ; Bees/microbiology ; Colony Collapse/microbiology ; Colony Collapse/prevention & control ; Disease Outbreaks ; Food Analysis ; Gram-Positive Bacterial Infections/diagnosis ; Gram-Positive Bacterial Infections/epidemiology ; Gram-Positive Bacterial Infections/prevention & control ; Honey/analysis ; Honey/microbiology ; Paenibacillus larvae/isolation & purification ; Paenibacillus larvae/physiology ; Saskatchewan/epidemiology ; Seasons ; Spores, Bacterial/isolation & purification
Chemical Substances	Anti-Bacterial Agents
Language	English
Publishing date	2022-02-07
Publishing country	United States
Document type	Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0263602
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Article ; Online: Are fungicides a driver of European foulbrood disease in honey bee colonies pollinating blueberries?

Jenna M. Thebeau / Allyssa Cloet / Dana Liebe / Fatima Masood / Ivanna V. Kozii / Colby D. Klein / Michael W. Zabrodski / Sarah Biganski / Igor Moshynskyy / Larhonda Sobchishin / Geoff Wilson / Maria Marta Guarna / Eric M. Gerbrandt / Antonio Ruzzini / Elemir Simko / Sarah C. Wood

Frontiers in Ecology and Evolution, Vol

2023 Volume 11

Abstract: IntroductionBlueberry producers in Canada depend heavily on pollination services provided by honey bees (Apis mellifera L.). Anecdotal reports indicate an increased incidence of European foulbrood (EFB), a bacterial disease caused by Melissococcus ... ...

Abstract	IntroductionBlueberry producers in Canada depend heavily on pollination services provided by honey bees (Apis mellifera L.). Anecdotal reports indicate an increased incidence of European foulbrood (EFB), a bacterial disease caused by Melissococcus plutonius, is compromising pollination services and colony health. Fungicidal products are commonly used in blueberry production to prevent fungal diseases such as anthracnose and botrytis fruit rot. Pesticide exposure has been implicated in honey bee immunosuppression; however, the effects of commercial fungicidal products, commonly used during blueberry pollination, on honey bee larval susceptibility to EFB have not been investigated.MethodsUsing an in vitro infection model of EFB, we infected first instar honey bee larvae with M. plutonius 2019 BC1, a strain isolated from an EFB outbreak in British Columbia, Canada, and chronically exposed larvae to environmentally relevant concentrations of fungicide products over 6 days. Survival was monitored until pupation or eclosion.ResultsWe found that larvae chronically exposed to one, two, or three fungicidal products [Supra® Captan 80WDG (Captan), low concentration of Kenja™ 400SC (Kenja), Luna® Tranquility (Luna), and/or Switch® 62.5 WG (Switch)], did not significantly reduce survival from EFB relative to infected controls. When larvae were exposed to four fungicide products concurrently, we observed a significant 24.2% decrease in survival from M. plutonius infection (p = 0.0038). Similarly, higher concentrations of Kenja significantly reduced larval survival by 24.7–33.0% from EFB (p < 0.0001).DiscussionThese in vitro results suggest that fungicides may contribute to larval susceptibility and response to M. plutonius infections. Further testing of other pesticide combinations is warranted as well as continued surveillance of pesticide residues in blueberry-pollinating colonies.
Keywords	pesticides ; fungicides ; European foulbrood ; honey bees (Apis mellifera) ; blueberries ; Evolution ; QH359-425 ; Ecology ; QH540-549.5
Language	English
Publishing date	2023-01-01T00:00:00Z
Publisher	Frontiers Media S.A.
Document type	Article ; Online
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Article ; Online: Tryptorubin A: A Polycyclic Peptide from a Fungus-Derived Streptomycete.

Wyche, Thomas P / Ruzzini, Antonio C / Schwab, Laura / Currie, Cameron R / Clardy, Jon

Journal of the American Chemical Society

2017 Volume 139, Issue 37, Page(s) 12899–12902

Abstract: Fungus-growing ants engage in complex symbiotic relationships with their fungal crop, specialized fungal pathogens, and bacteria that provide chemical defenses. In an effort to understand the evolutionary origins of this multilateral system, we ... ...

Abstract	Fungus-growing ants engage in complex symbiotic relationships with their fungal crop, specialized fungal pathogens, and bacteria that provide chemical defenses. In an effort to understand the evolutionary origins of this multilateral system, we investigated bacteria isolated from fungi. One bacterial strain (Streptomyces sp. CLI2509) from the bracket fungus Hymenochaete rubiginosa, produced an unusual peptide, tryptorubin A, which contains heteroaromatic links between side chains that give it a rigid polycyclic globular structure. The three-dimensional structure was determined by NMR and MS, including a
MeSH term(s)	Basidiomycota/chemistry ; Basidiomycota/metabolism ; Molecular Conformation ; Peptides, Cyclic/biosynthesis ; Peptides, Cyclic/chemistry ; Streptomyces/chemistry ; Streptomyces/metabolism
Chemical Substances	Peptides, Cyclic ; tryptorubin A
Language	English
Publishing date	2017-09-06
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	3155-0
ISSN	1520-5126 ; 0002-7863
ISSN (online)	1520-5126
ISSN	0002-7863
DOI	10.1021/jacs.7b06176
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Article: Investigation of Macrolide Resistance Genotypes in

Kinnear, Andrea / McAllister, Tim A / Zaheer, Rahat / Waldner, Matthew / Ruzzini, Antonio C / Andrés-Lasheras, Sara / Parker, Sarah / Hill, Janet E / Jelinski, Murray D

Pathogens (Basel, Switzerland)

2020 Volume 9, Issue 8

Abstract: Mycoplasma ... ...

Abstract	Mycoplasma bovis
Language	English
Publishing date	2020-07-30
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2695572-6
ISSN	2076-0817
ISSN	2076-0817
DOI	10.3390/pathogens9080622
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Article ; Online: Evaluating approved and alternative treatments against an oxytetracycline-resistant bacterium responsible for European foulbrood disease in honey bees

Fatima Masood / Jenna M. Thebeau / Allyssa Cloet / Ivanna V. Kozii / Michael W. Zabrodski / Sarah Biganski / Jenny Liang / M. Marta Guarna / Elemir Simko / Antonio Ruzzini / Sarah C. Wood

Scientific Reports, Vol 12, Iss 1, Pp 1-

2022 Volume 9

Abstract: Abstract European foulbrood (EFB) is a disease of honey bee larvae caused by Melissococcus plutonius. In North America, oxytetracycline (OTC) is approved to combat EFB disease though tylosin (TYL) and lincomycin (LMC) are also registered for use against ... ...

Abstract	Abstract European foulbrood (EFB) is a disease of honey bee larvae caused by Melissococcus plutonius. In North America, oxytetracycline (OTC) is approved to combat EFB disease though tylosin (TYL) and lincomycin (LMC) are also registered for use against American foulbrood disease. Herein, we report and characterize an OTC-resistant M. plutonius isolate from British Columbia, Canada, providing an antimicrobial sensitivity to the three approved antibiotics and studying their abilities to alter larval survival in an in vitro infection model. Specifically, we investigated OTC, TYL, and LMC as potential treatment options for EFB disease using laboratory-reared larvae infected with M. plutonius. The utility of the three antibiotics were compared through an experimental design that either mimicked metaphylaxis or antimicrobial intervention. At varying concentrations, all three antibiotics prevented clinical signs of EFB disease following infection with M. plutonius 2019BC1 in vitro. This included treatment with 100 μg/mL of OTC, a concentration that was ~ 3× the minimum inhibitory concentration measured to inhibit the strain in nutrient broth. Additionally, we noted high larval mortality in groups treated with doses of OTC corresponding to ~ 30× the dose required to eliminate bacterial growth in vitro. In contrast, TYL and LMC were not toxic to larvae at concentrations that exceed field use. As we continue to investigate antimicrobial resistance (AMR) profiles of M. plutonius from known EFB outbreaks, we expect a range of AMR phenotypes, reiterating the importance of expanding current therapeutic options along with alternative management practices to suppress this disease.
Keywords	Medicine ; R ; Science ; Q
Subject code	610
Language	English
Publishing date	2022-04-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: The catalytic serine of meta-cleavage product hydrolases is activated differently for C-O bond cleavage than for C-C bond cleavage.

Ruzzini, Antonio C / Horsman, Geoff P / Eltis, Lindsay D

Biochemistry

2012 Volume 51, Issue 29, Page(s) 5831–5840

Abstract: meta-Cleavage product (MCP) hydrolases catalyze C-C bond fission in the aerobic catabolism ... activated by the His-Asp dyad. In contrast, rapid acylation of the H265Q variant during C-C bond cleavage ... with a k(cat) value (6.3 ± 0.5 s(-1)) similar to that of HOPDA (6.5 ± 0.5 s(-1)). Consistent ...

Abstract	meta-Cleavage product (MCP) hydrolases catalyze C-C bond fission in the aerobic catabolism of aromatic compounds by bacteria. These enzymes utilize a Ser-His-Asp triad to catalyze hydrolysis via an acyl-enzyme intermediate. BphD, which catalyzes the hydrolysis of 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid (HOPDA) in biphenyl degradation, catalyzed the hydrolysis of an ester analogue, p-nitrophenyl benzoate (pNPB), with a k(cat) value (6.3 ± 0.5 s(-1)) similar to that of HOPDA (6.5 ± 0.5 s(-1)). Consistent with the breakdown of a shared intermediate, product analyses revealed that BphD catalyzed the methanolysis of both HOPDA and pNPB, partitioning the products to benzoic acid and methyl benzoate in similar ratios. Turnover of HOPDA was accelerated up to 4-fold in the presence of short, primary alcohols (methanol > ethanol > n-propanol), suggesting that deacylation is rate-limiting during catalysis. In the steady-state hydrolysis of HOPDA, k(cat)/K(m) values were independent of methanol concentration, while both k(cat) and K(m) values increased with methanol concentration. This result was consistent with a simple model of nucleophilic catalysis. Although the enzyme could not be saturated with pNPB at methanol concentrations of >250 mM, k(obs) values from the steady-state turnover of pNPB at low methanol concentrations were also consistent with a nucleophilic mechanism of catalysis. Finally, transient-state kinetic analysis of pNPB hydrolysis by BphD variants established that substitution of the catalytic His reduced the rate of acylation by more than 3 orders of magnitude. This suggests that for pNPB hydrolysis, the serine nucleophile is activated by the His-Asp dyad. In contrast, rapid acylation of the H265Q variant during C-C bond cleavage suggests that the serinate forms via a substrate-assisted mechanism. Overall, the data indicate that ester hydrolysis proceeds via the same acyl-enzyme intermediate as that of the physiological substrate but that the serine nucleophile is activated via a different mechanism.
MeSH term(s)	Benzoates/chemistry ; Benzoates/metabolism ; Burkholderia/chemistry ; Burkholderia/enzymology ; Burkholderia/metabolism ; Fatty Acids, Unsaturated/metabolism ; Hydrolases/chemistry ; Hydrolases/metabolism ; Hydrolysis ; Methanol/metabolism ; Serine/chemistry ; Serine/metabolism ; Substrate Specificity
Chemical Substances	Benzoates ; Fatty Acids, Unsaturated ; Serine (452VLY9402) ; 2-hydroxy-6-oxo-6-phenyl-2,4-hexadienoic acid (50480-67-6) ; Hydrolases (EC 3.-) ; 2,6-dioxo-6-phenylhexa-3-enoate hydrolase (EC 3.7.1.8) ; Methanol (Y4S76JWI15)
Language	English
Publishing date	2012-07-24
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1108-3
ISSN	1520-4995 ; 0006-2960
ISSN (online)	1520-4995
ISSN	0006-2960
DOI	10.1021/bi300663r
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