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Methods in
Molecular Biology 1918

Arnaud Bridier Editor

Foodborne
Bacterial
Pathogens
Methods and Protocols
METHODS IN MOLECULAR BIOLOGY

Series Editor
John M. Walker
School of Life and Medical Sciences
University of Hertfordshire
Hatfield, Hertfordshire, AL10 9AB, UK

For further volumes:

http://www.springer.com/series/7651
Foodborne Bacterial Pathogens

Methods and Protocols

Edited by

Arnaud Bridier
Fougères Laboratory, ANSES, Fougères, France
Editor
Arnaud Bridier
Fougères Laboratory
ANSES
Fougères, France

ISSN 1064-3745 ISSN 1940-6029 (electronic)

Methods in Molecular Biology
ISBN 978-1-4939-8999-7 ISBN 978-1-4939-9000-9 (eBook)
https://doi.org/10.1007/978-1-4939-9000-9
Library of Congress Control Number: 2018965210

© Springer Science+Business Media, LLC, part of Springer Nature 2019

This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is
concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction
on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation,
computer software, or by similar or dissimilar methodology now known or hereafter developed.
The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply,
even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations
and therefore free for general use.
The publisher, the authors, and the editors are safe to assume that the advice and information in this book are believed to
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This Humana Press imprint is published by the registered company Springer Science+Business Media, LLC, part of
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The registered company address is: 233 Spring Street, New York, NY 10013, U.S.A.
Preface

Ensuring food safety is a permanent challenge due to continuous evolutions of the produc-
tion scale and techniques, the globalization of food industry, and the desire to develop
sustainable food future. Despite the improvement of management policies and the imple-
mentation of regulatory standard to guarantee food safety and quality, foodborne diseases
remain an important cause of morbidity and mortality worldwide and have a significant
socioeconomic impact.
Improving the surveillance of bacterial pathogenic strains along the food chain, through
the improvement of detection, identification, and quantification methods, is crucial to
improve the microbiological quality of food and thus ensure the health of the consumers.
Moreover, a better deciphering of the ecology of foodborne pathogens and bacterial
strategies developed in response to food production conditions from farm to fork is abso-
lutely required to obtain a realistic vision of risk and to develop an efficient food safety
management through a One Health approach. Recently, the amazing improvements of
analytical methods and molecular biology technologies such as next-generation sequencing
(NGS) have provided access to novel and valuable data and enabled the development of
original integrative approaches feeding a holistic vision in this perspective. By bringing
together respected specialists in the field, this book constitutes a comprehensive collection
of cutting-edge methods, innovative approaches, and perspectives in the field of bacterial
foodborne pathogen analysis through three parts:
Part I. Bacterial Pathogen Detection and Quantification in Food
Part II. Phenotypic and Metabolic Characterization of Foodborne Pathogens
Part III. NGS and Modeling Approaches
The aim of the present book is to serve as a “field guide” both for researchers, students,
and food industrials who want to have an overview of current approaches and protocols used
to study bacterial foodborne pathogens. Due to the wide amplitude of the thematic
addressed here, this book does not claim to be an exhaustive collection of all methods
used to study bacterial foodborne pathogens but rather a compilation of various representa-
tive techniques and approaches currently used.
I am grateful to Prof. John Walker, chief editor of the Methods in Molecular Biology series
for the opportunity to edit this book and his assistance during the edition process. Finally, I
would like to express my sincere thanks to all the contributing authors for providing such
high-quality manuscripts.

Fougères, France Arnaud Bridier

v
Contents

Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix

PART I BACTERIAL PATHOGEN DETECTION AND QUANTIFICATION IN FOOD

1 Accelerated Sample Preparation for Fast Salmonella Detection
in Poultry Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Eduardo Ximenes, Seockmo Ku, Lori Hoagland, and Michael R. Ladisch
2 Direct or DNA Extraction-Free Amplification and Quantification
of Foodborne Pathogens. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
Maggie R. Williams and Syed A. Hashsham
3 The Use of Multiplex Real-Time PCR for the Simultaneous Detection
of Foodborne Bacterial Pathogens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Alejandro Garrido-Maestu, David Tomás Fornés
and Marta Prado Rodrı́guez
4 Sequence-Specific End Labeling of Oligonucleotides (SSELO)-Based
Microbial Detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Tanja Kostic and Levente Bodrossy
5 Rapid On-Site Detection and Quantification of Foodborne Pathogens
Using Microfluidic Devices. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Nobuyasu Yamaguchi
6 Modified Bacteriophage Tail Fiber Proteins for Labeling,
Immobilization, Capture, and Detection of Bacteria. . . . . . . . . . . . . . . . . . . . . . . . . 67
Matthew Dunne and Martin J. Loessner
7 EIS-Based Biosensors in Foodborne Pathogen Detection
with a Special Focus on Listeria monocytogenes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
Palmiro Poltronieri, Elisabetta Primiceri, and Rajeswaran Radhakrishnan

PART II PHENOTYPIC AND METABOLIC CHARACTERIZATION

OF FOODBORNE PATHOGENS

8 Method to Study the Survival Abilities of Foodborne Bacterial

Pathogens Under Food Processing Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105
Olivier Habimana
9 Viability Detection of Foodborne Bacterial Pathogens in Food
Environment by PMA-qPCR and by Microscopic Observation . . . . . . . . . . . . . . . 117
Thomas Brauge, Graziella Midelet-Bourdin, and Christophe Soumet
10 Isolation of Bacterial RNA from Foods Inoculated with Pathogens . . . . . . . . . . . 129
Deepti Tyagi, Autumn L. Kraft, and Teresa M. Bergholz

vii
viii Contents

11 Use of Two-Dimensional Electrophoresis to Explore Foodborne

Bacterial Pathogen Responses to Gastrointestinal Stress . . . . . . . . . . . . . . . . . . . . . 139
Denise Schrama and Maria Leonor Faleiro
12 Identification of Putative Biomarkers Specific to Foodborne Pathogens
Using Metabolomics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149
Snehal R. Jadhav, Rohan M. Shah, Avinash V. Karpe, David J. Beale,
Konstantinos A. Kouremenos, and Enzo A. Palombo
13 Characterization of Bacterial Membrane Fatty Acid Profiles
for Biofilm Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165
Florence Dubois-Brissonnet
14 High Content Screening Confocal Laser Microscopy
(HCS-CLM) to Characterize Biofilm 4D Structural Dynamic
of Foodborne Pathogens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171
Alexis Canette, Julien Deschamps, and Romain Briandet
15 Static Immersion and Injection Methods for Live Cell Imaging
of Foodborne Pathogen Infections in Zebrafish Larvae . . . . . . . . . . . . . . . . . . . . . . 183
Macarena A. Varas, Javiera Ortı́z-Severı́n, Andrés E. Marcoleta,
Carlos A. Santiviago, Miguel L. Allende, and Francisco P. Chávez
16 Use of C. elegans Diapause to Study Transgenerational Responses
to Pathogen Infection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191
Francisco P. Chávez and Andrea Calixto

PART III NGS AND MODELING APPROACHES

17 Utilizing the Public GenomeTrakr Database for Foodborne

Pathogen Traceback. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 201
Ruth E. Timme, Maria Sanchez Leon, and Marc W. Allard
18 Establishment of a Standardized 16S rDNA Library
Preparation to Enable Analysis of Microbiome in Poultry
Processing Using Illumina MiSeq Platform . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213
Kristina M. Feye and Steven C. Ricke
19 Exploring Foodborne Pathogen Ecology and Antimicrobial Resistance
in the Light of Shotgun Metagenomics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229
Arnaud Bridier
20 Modeling Growth of Listeria and Lactic Acid Bacteria
in Food Environments. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 247
Paw Dalgaard and Ole Mejlholm
21 Network Biology Approaches to Identify Molecular
and Systems-Level Differences Between Salmonella Pathovars . . . . . . . . . . . . . . . . 265
Marton Olbei, Robert A. Kingsley, Tamas Korcsmaros,
and Padhmanand Sudhakar

Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 275
Contributors

MARC W. ALLARD  Center for Food Safety and Applied Nutrition, U.S. Food and Drug
Administration, College Park, MD, USA
MIGUEL L. ALLENDE  Facultad de Ciencias, Centro FONDAP de Regulacion del Genoma,
Universidad de Chile, Santiago, Chile
DAVID J. BEALE  Land and Water, Commonwealth Scientific and Industrial Research
Organisation (CSIRO), Brisbane, QLD, Australia
TERESA M. BERGHOLZ  Department of Microbiological Sciences, North Dakota State
University, Fargo, ND, USA
LEVENTE BODROSSY  Environmental Genomics Team, CSIRO Oceans and Atmosphere,
Hobart, TAS, Australia
THOMAS BRAUGE  Laboratory for Food Safety, French Agency for Food, Environmental and
Occupational Health and Safety, Boulogne sur Mer, France; RMT Chlean, Joint
Technological Network: Hygienic Design of Production Lines and Equipment, France
ROMAIN BRIANDET  Micalis Institute, INRA, AgroParisTech, Université Paris-Saclay, Jouy-
en-Josas, France
ARNAUD BRIDIER  Fougères Laboratory, ANSES, Fougères, France
ANDREA CALIXTO  Facultad de Ciencias, Centro de Genomica y Bioinformática,
Universidad Mayor, Santiago, Chile; Facultad de Ciencias, Centro Interdisciplinario de
Neurociencia de Valparaı́so, Universidad de Valparaı́so, Valparaiso, Chile
ALEXIS CANETTE  Micalis Institute, INRA, AgroParisTech, Université Paris-Saclay, Jouy-en-
Josas, France; IBPS Institute, Sorbonne Université, CNRS, INSERM, Paris, France
FRANCISCO P. CHÁVEZ  Laboratorio de Microbiologı́a de Sistemas, Departamento de Biologı́a,
Facultad de Ciencias, Universidad de Chile, Santiago, Chile
PAW DALGAARD  Food Microbiology and Hygiene (Research Group), Division of Microbiology
and Production, National Food Institute (DTU Food), Technical University of Denmark
(DTU), Kongens Lyngby, Denmark
JULIEN DESCHAMPS  Micalis Institute, INRA, AgroParisTech, Université Paris-Saclay, Jouy-
en-Josas, France
FLORENCE DUBOIS-BRISSONNET  UMR Micalis, INRA, AgroParisTech, Université Paris-
Saclay, Jouy-en-Josas, France
MATTHEW DUNNE  Laboratory of Food Microbiology, Institute of Food, Nutrition and
Health, ETH Zurich, Zurich, Switzerland
MARIA LEONOR FALEIRO  CMBR, University of Algarve, Faro, Portugal
KRISTINA M. FEYE  Department of Food Science, Center for Food Safety, University of
Arkansas, Fayetteville, AR, USA
ALEJANDRO GARRIDO-MAESTU  Department of Life Sciences, Food Quality and Safety
Research Group, International Iberian Nanotechnology Laboratory, Braga, Portugal
OLIVIER HABIMANA  School of Biological Sciences, The University of Hong Kong, Hong Kong,
SAR, People’s Republic of China
SYED A. HASHSHAM  Department of Civil and Environmental Engineering, Michigan State
University, East Lansing, MI, USA; Center for Microbial Ecology, Michigan State
University, East Lansing, MI, USA

ix
x Contributors

LORI HOAGLAND  Department of Horticulture and Landscape Architecture, Purdue

University, West Lafayette, IN, USA
SNEHAL R. JADHAV  Department of Chemistry and Biotechnology, School of Science,
Swinburne University of Technology, Melbourne, VIC, Australia; Centre for Advanced
Sensory Science, School of Exercise and Nutrition Sciences, Deakin University, Melbourne,
VIC, Australia
AVINASH V. KARPE  Department of Chemistry and Biotechnology, School of Science,
Swinburne University of Technology, Melbourne, VIC, Australia; Land and Water,
Commonwealth Scientific and Industrial Research Organisation (CSIRO), Brisbane,
QLD, Australia
ROBERT A. KINGSLEY  Quadram Institute Bioscience, Norwich Research Park, Norwich, UK
TAMAS KORCSMAROS  Quadram Institute Bioscience, Norwich Research Park, Norwich, UK;
Earlham Institute, Norwich Research Park, Norwich, UK
TANJA KOSTIC  Bioresources Unit, AIT Austrian Institute of Technology, Tulln an der
Donau, Austria
KONSTANTINOS A. KOUREMENOS  Metabolomics Australia, Bio21 Molecular Science and
Biotechnology Institute, The University of Melbourne, Melbourne, VIC, Australia
AUTUMN L. KRAFT  Department of Microbiological Sciences, North Dakota State University,
Fargo, ND, USA
SEOCKMO KU  Fermentation Science Program, School of Agribusiness and Agriscience,
College of Basic and Applied Sciences, Middle Tennessee State University, Murfreesboro,
TN, USA
MICHAEL R. LADISCH  Department of Agricultural and Biological Engineering, Purdue
University, West Lafayette, IN, USA; Laboratory of Renewable Resources Engineering,
Purdue University, West Lafayette, IN, USA; Weldon School of Biomedical Engineering,
Purdue University, West Lafayette, IN, USA
MARTIN J. LOESSNER  Laboratory of Food Microbiology, Institute of Food, Nutrition and
Health, ETH Zurich, Zurich, Switzerland
ANDRÉS E. MARCOLETA  Laboratorio de Biologı́a Estructural y Molecular, Departmento de
Biologı́a, Facultad de Ciencias, Universidad de Chile, Santiago, Chile
OLE MEJLHOLM  Corporate Quality, Royal Greenland Ltd., Svenstrup J, Denmark
GRAZIELLA MIDELET-BOURDIN  Laboratory for Food Safety, French Agency for Food,
Environmental and Occupational Health and Safety, Boulogne sur Mer, France; RMT
Chlean, Joint Technological Network: Hygienic Design of Production Lines and
Equipment, France
MARTON OLBEI  Quadram Institute Bioscience, Norwich Research Park, Norwich, UK;
Earlham Institute, Norwich Research Park, Norwich, UK
JAVIERA ORTÍZ-SEVERÍN  Laboratorio de Microbiologı́a de Sistemas, Departamento de
Biologı́a, Facultad de Ciencias, Universidad de Chile, Santiago, Chile
ENZO A. PALOMBO  Department of Chemistry and Biotechnology, School of Science,
Swinburne University of Technology, Melbourne, VIC, Australia
PALMIRO POLTRONIERI  CNR-ISPA, Lecce, Italy
MARTA PRADO RODRÍGUEZ  Department of Life Sciences, Food Quality and Safety Research
Group, International Iberian Nanotechnology Laboratory, Braga, Portugal
ELISABETTA PRIMICERI  CNR-Nanotec, Lecce, Italy
RAJESWARAN RADHAKRISHNAN  Faraday Technologies, Clayton, OH, USA
STEVEN C. RICKE  Department of Food Science, Center for Food Safety, University of
Arkansas, Fayetteville, AR, USA
 Contributors xi

MARIA SANCHEZ LEON  Center for Food Safety and Applied Nutrition, U.S. Food and Drug
Administration, College Park, MD, USA
CARLOS A. SANTIVIAGO  Laboratorio de Microbiologı́a, Departamento de Bioquı́mica y
Biologı́a Molecular, Facultad de Ciencias Quı́micas y Farmacéuticas, Universidad de
Chile, Santiago, Chile
DENISE SCHRAMA  CCMAR, University of Algarve, Faro, Portugal
ROHAN M. SHAH  Department of Chemistry and Biotechnology, School of Science, Swinburne
University of Technology, Melbourne, VIC, Australia
CHRISTOPHE SOUMET  RMT Chlean, Joint Technological Network: Hygienic Design of
Production Lines and Equipment, France; Fougères Laboratory, ANSES, Fougères, France
PADHMANAND SUDHAKAR  Quadram Institute Bioscience, Norwich Research Park, Norwich,
UK; Earlham Institute, Norwich Research Park, Norwich, UK
RUTH E. TIMME  Center for Food Safety and Applied Nutrition, U.S. Food and Drug
Administration, College Park, MD, USA
DAVID TOMÁS FORNÉS  Nestlé Research Center, Institute Food Safety and Analytical Science,
Microbial and Molecular Analytics Group, Lausanne, Switzerland
DEEPTI TYAGI  Department of Microbiological Sciences, North Dakota State University,
Fargo, ND, USA
MACARENA A. VARAS  Laboratorio de Microbiologı́a de Sistemas, Departamento de Biologı́a,
Facultad de Ciencias, Universidad de Chile, Santiago, Chile; Laboratorio de Biologı́a
Estructural y Molecular, Departmento de Biologı́a, Facultad de Ciencias, Universidad de
Chile, Santiago, Chile
MAGGIE R. WILLIAMS  Department of Civil and Environmental Engineering, Michigan
State University, East Lansing, MI, USA
EDUARDO XIMENES  Department of Agricultural and Biological Engineering, Purdue
University, West Lafayette, IN, USA; Laboratory of Renewable Resources Engineering,
Purdue University, West Lafayette, IN, USA
NOBUYASU YAMAGUCHI  Osaka Institute of Public Health, Osaka, Japan
 Part I

Bacterial Pathogen Detection and Quantification in Food

 Chapter 1

Accelerated Sample Preparation for Fast Salmonella

Detection in Poultry Products
Eduardo Ximenes, Seockmo Ku, Lori Hoagland, and Michael R. Ladisch

Abstract
Salmonella is the most burdensome foodborne pathogen in the USA and a major causal agent of foodborne
outbreaks. Detection of a pathogen such as Salmonella can be achieved within a few hours using commer-
cially available rapid methods, but the sample preparation is time consuming and may require multiple days.
We have developed and successfully tested an accelerated sample preparation method based on microfiltra-
tion, in some cases preceded by a short enrichment step, for the rapid detection of selected pathogens. The
time-frame of the overall process, from sample preparation (i.e., food rinse or homogenate preparation,
microbial enrichment, and filtration steps) to detection is 8 h or less. While microfiltration has been
practiced for 70 years, the complex interactions between food substances and filter membrane surfaces
have shown that food pretreatment methods need to be developed on a case by case basis for the recovery of
bacteria from food homogenates and/or rinses. We have also demonstrated that addition of protease to
treat homogenates of different poultry products prior to microfiltration avoids the rapid decrease in flux
that otherwise occurs during microfiltration. This protease treatment minimizes filter clogging, so that the
microbial concentration, recovery and detection of 1 to 10 CFU/g of Salmonella in poultry products is
possible in less than 8 h.

Key words Sample preparation, Salmonella detection, Poultry products, Microfiltration, Hollow fiber
membranes, Protease

1 Introduction

The detection of a pathogen such as Salmonella can be achieved

within a few hours using commercially available rapid methods
(e.g., immunoassays and molecular methods). The sample prepara-
tion step, however, is time-consuming and may take multiple days
[1–4]. We have developed an accelerated sample preparation meth-
odology in our laboratory [5], which was initially tested for poultry
products as described here. This approach combines short enrich-
ment of microorganisms originally present at low numbers
(
1 CFU/g) with enzyme hydrolysis and a prefiltration step fol-
lowed by hollow-fiber microfiltration and polymerase chain

Arnaud Bridier (ed.), Foodborne Bacterial Pathogens: Methods and Protocols, Methods in Molecular Biology, vol. 1918,
https://doi.org/10.1007/978-1-4939-9000-9_1, © Springer Science+Business Media, LLC, part of Springer Nature 2019

3
4 Eduardo Ximenes et al.

reaction (PCR) for the rapid detection of target foodborne patho-

gens. The overall approach takes 8 h or less [3, 6–8].
Pathogenic microorganisms may pass through the food chain
and be transmitted to humans using different type of foods, includ-
ing beef, poultry, leafy greens, and eggs as major vehicles
[9]. Microbial contamination of food, such as poultry related pro-
ducts, can occur throughout the production line, affecting the
quality of the final product [10].
Nontyphoidal salmonellosis causes an estimated 1.2 million
illnesses, 19,000 hospitalizations, and 370 deaths annually, being
the most common enteric pathogenic disease in the USA
[11, 12]. The majority of Salmonella spp. infections are foodborne,
where 11% incidences are due to contact with animals or zoonosis
[12, 13].
Reptiles, amphibians, live poultry, and small nontraditional pets
are at a high risk for zoonotic salmonellosis [12–17]. Infected
animals, while often appearing healthy, can intermittently shed
bacteria [12, 16]. Zoonotic Salmonella infection can occur not
only through direct contact with an infected animal but also via
exposure to the place or objects where the animals carrying Salmo-
nella live. Furthermore, infection may occur by ingesting food or
drink prepared in a polluted environment [12, 13, 17, 18]. It is
worth noting that backyard poultry flocks are increasing in popu-
larity in the USA as a consequence of the local food movement and
the public’s desire to raise live poultry for fun or as a hobby [19]. As
a result, poultry farming has become an emerging public health
issue that causes additional salmonellosis outbreaks [12, 17].
Pathogens can form biofilms making their detection more
challenging [20]. Biofilms are cell (bacteria, fungi, algae, or proto-
zoa) aggregates contained in self-produced extracellular polymers
that facilitate: (1) attachment onto biological or abiotic surfaces;
(2) alteration of microbial growth rates; and (3) gene transcription
and translation [21, 22]. While the matrix (extracellular material)
accounts for more than 90% of the total biofilm composition, the
total mass of the microorganisms is less than 10% of the dry mass.
This matrix is mainly produced by microorganisms present in the
biofilm and is composed of various biopolymers (also known as
extracellular polymers [EPS]). The EPS acts as the skeleton of the
three-dimensional biofilm structure and participates in the surface
attachment to biological or abiotic materials and biofilm agglomer-
ation [23, 24]. Biofilm protects microorganisms from natural ene-
mies (e.g., bacteriophages and amoebas) as well as the various
biocides and physical treatments (desiccation) used in food proces-
sing [22, 25]. Significant consumer protection will be achieved by
the further understanding of the factors that allow human patho-
gens to survive on poultry products complemented by more accu-
rate and rapid sample preparation and detection protocols.
 Accelerated Food Sample Preparation 5

Microfiltration is a conceptually simple approach to bring large

food samples (homogenates and rinses) to a smaller volume, while
also increasing cell concentration, avoiding lengthy standard cul-
turing and/or enrichment processes [3, 6–8]. Although it has been
recognized more than 40 years ago by Sharp et al. [26] that food
can be filtered in quantities pertinent to the maximums used in
conventional plating procedures, there were still hurdles to over-
come for the development of a rapid filtration approach aiming
microorganism concentration, recovery, and detection in foods
[3, 6–8].
Cross-flow microfiltration (also known as tangential flow) is a
potentially effective approach for the cases where the suspension to
be filtered contains fine particles or microbial suspensions, the
density is close to that of the suspending fluid and the particles
have the tendency to form stable and compressible protein fats, oils
deposits or films [3, 27]. The hollow-fiber modules used for the
cross-flow microfiltration has high surface area-to-volume and large
permeate flux rates per filter unit volume. After the concentration
step, the membrane is backflushed for the recovery of concentrated
cells in an aqueous buffer or flushed with an elution buffer through
the fiber to recover cells for detection using conventional plating or
fast detection methods [28]. Hollow-fiber membranes have been
demonstrated for pathogen concentration and recovery from water
samples [29–32], but for large volumes of aqueous food extracts,
where rapid membrane plugging occurs, have proved to be more
challenging for this purpose [3]. Unlike water, the food homoge-
nates has a heterogeneous nature, consisting of naturally occurring
microbiota and varied compounds (carbohydrates, proteins, lipids,
micron-sized particulates, and inorganic food components) as part
of the matrix. Proteins are known to foul microfiltration mem-
branes [6, 33, 34].
We have reported the discovery and development of the use of
enzymes to minimize membrane fouling with little, if any, impact
on the viability of the target microorganisms being isolated
[3, 5–8]. To this end, we have added proteases to stomached and
filtered solutions of chicken, chicken carcass rinse, egg white, and
Turkey burger [3, 5–8], so that 250–500 mL extracts could be
processed into 1–5 mL final volume samples within an hour using
0.3–0.5 mm diameter modified polyethersulfone (mPES) hollow-
fiber membrane modules [5]. Hydrodynamics of the fiber module
allows to control membrane fouling, where fluid flow rates on both,
the permeate (outside) and retentate (inside) parts of the mem-
branes, are controlled using two pumps. Constant transmembrane
pressure drops is maintained by automated control of these pumps
[6]. Other research has resulted in protocols to sterilize the pumps,
connective tubing, and hollow fiber membranes between uses. In
this way, each hollow fiber module can be used multiple times.
6 Eduardo Ximenes et al.

Regular chemical cleaning is required to ensure reproducible

performance and repeated membrane use, even though tangential
flow can reduce the rate of the buildup of foulants on the mem-
brane surface [35]. Therefore, a hydraulic and chemical cleaning
procedure using deionized (DI) water, NaOH, and 70% of ethanol
is performed between consecutive runs of sample processing, also
aiming system sterilization between samples. This procedure helps
to reduce the cost of the membrane module, taking in consider-
ation that more samples can be processed before the membrane
need to be replaced [3, 5–8].

2 Materials

2.1 Reagents 1. Growth media (BBL™ Brain Heart Infusion (BHI) agar,
CHROMagar™ Salmonella, Xylose lysine deoxycholate
(XLD) agar, Rappaport Vassiliadis (RV) broth).
2. Sterilized distilled (DI) water.
3. Double-distilled water (ddH2O).
4. Buffers (Buffered peptone water, Phosphate buffered saline
(PBS supplemented with 0.002% (v/v) Tween 20).
5. Cleaning and sterilization reagents (10% (v/v) bleach, 70%
(v/v) ethanol, 0.2 M NaOH).
6. Commercial available enzymes (see Note 1).
7. DNA isolation commercially available kits.
8. Primers for qPCR and conventional PCR reactions:
– invA forward 50 -GTGAAATAATCGCCACGTCGGCAA-
30 and invA reverse 50 -TCATCGCACCGTCAAAGGA
ACC-30
– hilA forward 50 -CTGCCGC AGTGTTAAGGATA-30 and
hilA reverse: 50 -GTCGCCTTAATC GCATGG-30,
– hisJ forward 50-ACTGGCGTTATCCCTTT CTCTGGT
G-30 and hisJ reverse 50 -GTTGTCCTGCCCCT GG TAA-
GAGA-30 .
9. SYBR green PCR master mix for qPCR.
10. Deoxynucleotide mix.
11. Standard Taq reaction buffer: 50 mM KCl, 10 mM Tris
pH 9.0, 0.1% Triton X-100, 2 mM MgCl2 (final
concentration).
12. Taq polymerase.
13. Gel electrophoresis reagents (agarose and ethidium bromide).
 Accelerated Food Sample Preparation 7

2.2 Equipment 1. A stomacher and sterile bags with filter.

2. Incubator and refrigerator at 37  C and 4  C, respectively.
3. A blender and sterile blender bags with 500 μm filters.
4. Continuous Cell Concentration Device (C3D) with commer-
cial polyethersulfone hollow fiber membrane.
5. Personal computer (PC) with an operational control and data
acquisition conducted via graphical interface in a program built
with LabVIEW 2009.
6. Peristaltic and vacuum pumps.
7. 2.7 μm cutoff borosilicate GF/D glass membrane.
8. Commercial polyethersulfone hollow fiber membrane with
0.2 μm pore size. The module specifications are as follows:
140 cm2 surface area, 0.50 mm inner diameter, 20 cm length,
and a nominal pore cutoff of 0.2 mm with 45 fibers.
9. PCR instrument (Thermal cycling and BAX® System PCR
(DuPont Qualicon).
10. UV transilluminator.
11. A spectrophotometer for DNA concentration determination.

3 Methods

The protocols here are based on the work of Li et al., Vibbert et al.,
Ku et al., and Ladisch and Ximenes [3, 5–8]. All of the steps below
should be conducted under aseptic conditions. The entire proce-
dure, from the sample processing to detection (by PCR and other
rapid detection methods) of Salmonella, can be achieved in less
than 8 h (Fig. 1).
For food homogenate preparation we have followed the US
Department of Agriculture’s Food Safety and Inspection Service
Microbiology Laboratory Guidebook [36], the Food and Drug
Administration(FDA)’s Bacteriological Analytical Manual (BAM)
[37], and International Organization for Standardization(ISO)‘s
6579:2002 method (Microbiology of food and animal feed stuffs:
horizontal method for the detection of Salmonella spp.) [38]
guidelines with the modifications indicated below for specific
foods. A typical example of results obtained for the overall proce-
dure with the approach reported here is shown in Fig. 2.
8 Eduardo Ximenes et al.

Fig. 1 Schematic overview of accelerated sample preparation for fast Salmonella

detection in poultry products

250 14

12
200

Salmonella (log CFU/mL)

10
Sample volume (mL)

150
8

6
100

50
2

0 0
Original Enrichment GF/D Enzyme C3D microfuge
Volume, mL 210±10 210±10 202±25 202±25 4±1.7 0.4±0.1
Cell,log CFU/ mL 1.3±1.5 3.2±3.5 3.1±3.5 3.5±3.7 4.1±4.3 6.0±6.1
Time for each step, h 0.17 3 0.25 0.33 1.33 0.17

Fig. 2 Decreased total sample volumes (□) and increased Salmonella concentration (Δ) during the process of
cell concentration and recovery from ground turkey homogenates. The data represents mean values of
12 experiments (Ku et al., 2017, permission will be requested)
 Accelerated Food Sample Preparation 9

3.1 Preparation of 1. For chicken homogenates, mix 25 g of thin-cut chicken flesh

Chicken Homogenates and skin (from the grocery store) with 250 mL of sterile
and Chicken Carcass deionized water in a sterile Filtra Bag.
Rinsates (for the 2. Homogenize it in a Stomacher at 100 rpm for 30 s [3].
Latter, Based on 3. For chicken carcass rinsates, mix the chicken carcass with
Laboratory 400 mL of sterile buffered peptone water (BPW) in a sterile
Guidebook’s Method bag at room temperature by inverting the carcass back and
Number 4.09 [36]): forth for 1 min [6] (see Note 2).

3.2 Preparation of 1. Sterilize a blender between uses by soaking it in 10% (v/v)

Egg White bleach for 10 min and then 70% (v/v) ethanol overnight at
Homogenates room temperature.
(Modified Standard ISO 2. Soak four fresh whole eggs (grade A, from the grocery store) in
6579:2002 Method) 70% (v/v) ethanol for 30 min at room temperature and air-dry
[38]: them in a sterile hood to prevent sample contamination
[37, 39].
3. Broke the shells using a sterile spoon. After removing the egg
yolk using the sterile spoon, homogenize 100 g egg whites in a
blender for 15 s.
4. Prepare the aqueous egg white homogenates by mixing 25 g
homogenized egg whites and 500 mL BPW (approximately
pH 7.0) [7].

3.3 Preparation of 1. Transfer 25 g sample of ground Turkey (we have tested 93/7%
Ground Turkey of lean protein/fat from grocery store) to a sterilized plastic
Hamburger Samples bag and press by hand from outside the bag into a roughly 1 cm
thick square. Let the sample to be brought to room tempera-
ture. The square is then folded along its median, and gently
massaged by hand back to the original 1 cm thickness [40].
2. After folding and massaging is repeated one more time, place
the ground Turkey in blender bags) and mix with 225 mL
Rappaport Vassiliadis (RV) broth (instead of water or BPW in
order to favor Salmonella growth in the presence of a high
number of naturally occurring microorganisms, see Note 3).
3. Homogenize the resulting sample in a stomacher at 100 rpm
for 30 s.
4. Transfer the aqueous fraction to a 500 mL sterilized bottle for
the following steps.

3.4 Salmonella 1. Spike Salmonella cells at the desired sample concentration,

Inoculation (See either before (in chicken and ground Turkey samples, the
Note 4) microorganism is spread on the food surface) [3, 6, 7] or
during the preparation of the food homogenates (eggs samples
[8]). For eggs, after removing the egg yolk using a sterile spoon
(refer to previous section), 100 g egg whites is homogenized in
a blender for 15 s, and then artificially spiked with Salmonella.
10 Eduardo Ximenes et al.

2. The artificial cell inoculation can alternatively be performed

after the prefiltration step.

3.5 Enzyme 1. Perform enzyme reaction at conditions of 37  C and 200 rpm.

Treatment (See Note 5) While these are not optimal protease reaction conditions, they
and Prefiltration are effective for the desired effect of protein degradation with-
out affecting the microbial viability (see Note 5). The incuba-
tion time can be adjusted in accordance to the conditions of the
experiment (see Note 6).
2. For prefiltration (see Note 7) of chicken homogenates/rinses
and ground Turkey (see Note 8 for the latter) using a vacuum
pump, filter the samples using a 2.7 mm cutoff borosilicate
GF/D glass membrane (see Note 9). The spaces between the
fibers are small enough, and the depth of the disk is sufficiently
thick to remove the particles, while large enough to allow the
microorganisms to flow through.
3. In the case of egg white homogenates, there is no need to
perform a prefiltration step, once only the enzyme treatment
with proteases is needed to generate a sample that will not
cause fouling of the microfiltration membrane in the following
step [7].

3.6 Microfiltration 1. Concentration and recovery of cells uses a continuous cell

for Salmonella concentration device (C3D, see Notes 10 and 11), where the
Concentration and initial larger volume (250–500 mL) of the food samples con-
Recovery taining the microorganisms is reduced to a smaller volume
(about 5 mL), using a polyethersulfone hollow fiber membrane
with 0.2 μm pore size.
2. Elute the cells at the end of the cycle, loading 10 mL of DI
water or PBS (supplemented with 0.002% (v/v) Tween 20) in
the C3D.
3. Run the cell concentration mode after the speeds of pumps
1 and 2 are adjusted to 100% and 20% of their maximums,
respectively, resulting in a laminar cross-flow velocity of
0.375 m/s (corresponding to a Reynolds number of 117).
Cell recovery occurs when the retentate is eluted into a sample
collection tube (see Notes 11 and 12), instead of being circu-
lated back to the sample reservoir. Pump 1 is used for this step
and set to its maximum speed [3, 5–8].

3.7 C3D Cleaning 1. Follow the sample concentration with an immediate system
(See Note 13) rinsing and washing using sterilized DI water to detach the
weakly bound sample residues inside the system tubing, and
attenuate the surface layer of precipitates [41, 42].
2. After rinsing, treat the instrument with a multistep chemical
cleaning process developed as part of this methodology, as
 Accelerated Food Sample Preparation 11

follows: (1) NaOH has the ability to saponify the fat and
dissolve the protein particles to some extent. As the first step,
a 0.2 M NaOH solution is passed through the rig and allowed
to incubate inside of the system for 5 min, before removed for
the next step being performed; (2) this is followed by feeding
and flushing the system with sterilized DI water to remove the
residual NaOH inside the instrument; (3) 70% (v/v) ethanol is
fed into the system via the rig to return the system to aseptic
conditions; (4) the final step is feeding and flushing the system
with sterilized DI water to remove the residual ethanol inside
the system and rehydrate the filter membrane.
3. Perform all of the above steps with flow paths for both the
sample concentration and microbial recovery modes. For this
purpose, system pumps 1 and 2 are adjusted to 100% and 20%
of their maximum speeds, respectively. Both the permeate and
retentate are discarded in a waste reservoir. The membrane and
fluid contact area of the instrument are compatible with the
chemicals used.
4. Collect a sample of the sterilized DI water added in the final
step, and plate, as described in the next section, to verify the
sterilization of the instrument for reusing.

3.8 Salmonella 1. Enumerate the total number of cells on BBL BHI agar, while
Detection selectively enumerating the number of Salmonella cells by
plating on selective media (CHROMagar Salmonella or Xylose
3.8.1 Plating
lysine deoxycholate [XLD].
2. Incubate the plates at 37  C for 24 h. It may take longer for the
colonies to develop on the selective media, and in this case
longer incubation time (36–48 h) may be needed.
3. Express the cell concentration as log CFU/mL and calculate
cell concentration and recovery efficiency using Eqs. 1 and 2,
respectively.

Concentration factor ¼ ðlogcell concentration in concentrated sample=

ð1Þ
cell concentration in prefiltered sampleÞ
Recovery ð%Þ ¼ ðcell concentration  volume of concentrated sample=
ð2Þ
cell concentration  volume of prefiltered sampleÞ  100
where cell concentrations are in CFU/mL and volume is
in mL.

3.8.2 qPCR 1. Isolate DNA using commercially available kits according to the
manufacturer’s instructions.
2. Determine the concentration of isolated DNA from microor-
ganisms using a spectrophotometer.
12 Eduardo Ximenes et al.

3. Dilute the isolated DNA from different bacterial species in

ddH2O to 1 ng/μL for specificity testing, and perform qPCR.
4. Prepare the genomic DNA standard for real-time quantifica-
tion by serially diluting (1:10) the DNA of the pure cultures in
ddH2O. The copy number of the invA gene per ng is calcu-
lated based on the molecular weight of the 4685-kbp genome
of Salmonella Enteritidis [43]), where 1 ng of DNA corre-
sponded to 2  105 genome or bacterial cell equivalents. The
number of bacterial cell equivalents (BCE) in a sample is then
extrapolated from the qPCR signal of the standard [3].
5. Carry out SYBR green qPCR analysis of the samples using
species-specific primers for the invA gene in Salmonella enter-
ica (see Note 14).
6. Prepare the PCR mixture by mixing 10 μL of 2 SYBR green
PCR master mix, 0.3 μM of each primer, and 2 μL of DNA
template and nuclease-free water for a total volume of 20 μL.
All runs include a negative control without template DNA and
Salmonella Enteritidis PT21 standards to obtain the standard
curve.
7. Adjust the thermal cycling conditions as follows: 15 min at
95  C followed by 40 cycles; 1 cycle of 15 s at 95  C and 15 s
at 68  C; and a final melt cycle of 15 s at 95  C and 60 s at 60  C
with temperature increments of 0.3  C. All qPCR analyses are
performed in triplicate. The reproducibility of SYBR green
qPCR is confirmed by independently running samples on dif-
ferent days [3].

3.8.3 Conventional PCR 1. Prepare Salmonella crude lysates for PCR analysis after
heating at 95  C for 15 min. This is followed by Salmonella
DNA separation using a commercial DNA extraction kit (see
Note 15).
2. Perform the amplification reactions in a final volume of 20 μL,
containing 2 μL bacterial DNA, 200 μmol deoxynucleotide
mix, standard Taq reaction buffer, 1.5 units of Taq polymerase,
and 5 pmol of invA, hilA, and hisJ forward and reverse primers
tested individually or in combination.
3. Conduct the PCR amplification using a thermal cycler as fol-
lows: 95  C for 1 min, followed by 35 cycles at 94  C for 30 s,
62  C for 30 s, 72  C for 30 s, a final extension step at 72  C for
2 min, for a total time of 124 min. A negative control DNA
template of water and Salmonella-free food homogenate is also
included. The amplification products are resolved through
electrophoresis on a 2.0% (w/v) agarose gel and visualized
using ethidium bromide staining and a UV transilluminator
[5–8].
 Accelerated Food Sample Preparation 13

3.9 BAX® System The assay is performed according to the manufacturer’s instruc-
PCR Assay tions for Salmonella detection.

4 Notes

1. Commercial enzyme preparations that can be used include:

Protex 7L, 1600 azo units/g, (from Genencor Division of
Danisco, Rochester, NY, USA); PromodTM 298 L, 150 azo
units/g; and Promod 439L, 220 casein protease U/g (from
Biocatalysts (Wales, UK).
2. The wash water includes blood, as evidenced from the red
color. The extract is protein buffered at pH 6.8 [6, 44].
3. RV selective medium can be successfully used to increase the
Salmonella population in ground Turkey (3 h incubation at
37  C and 200 rpm) and decrease the number of naturally
occurring background microorganisms other than Salmonella
[8]. This is also an option for other type of foods and/or
microorganisms, especially when the later are expected to be
present at very low original levels (
1 CFU/g).
4. Salmonella enterica serovar Enteritidis phage type (PT) 21
stocks used in all of our tests were obtained from Dr. Arun
Bhunia’s Molecular Food Microbiology laboratory at Purdue
University (West Lafayette). The doubling time for Salmonella
can be estimated using the equation td ¼ t/(3.3(log X/X0)),
where td is the doubling time, t is the time period of cell
growth, X is the number of Salmonella at time t, and X0 is
the number of Salmonella at the start time [45].
5. For the sample concentration using microfiltration, the mem-
brane pore size must be small enough to retain the microor-
ganisms and large enough to allow small particles and soluble
materials to move into the permeate side, generating a concen-
trated microorganism sample. Because of fouling by proteins
due to film formation at the membrane surface or travel into
pores and aggregate formation that plugs the membrane, even
if the membrane have a relatively large pore size, i.e.,
0.20–0.45 μm, it may not perform as anticipated [6]. This
will result in the flux through the membrane quickly decreased
in the presence of some proteins [33, 34]. Our work has shown
that proteases minimize HF membrane fouling, and do not
affect microbial viability (Fig. 3) [6–8]. We have performed
selective enrichment in the presence of such enzymes for 2 h or
more without any noticeable effect on cell viability [8]. Salmo-
nella cells used in our tests are stable when incubated with
protease in buffered peptone water or poultry products sam-
ples. However, in the absence of protein or peptide, we
14 Eduardo Ximenes et al.

0.4

0
y = 0.0016x - 0.0354
2
R = 0.2338

Log (X/X0)
-0.4

-0.8
y = -0.0305x + 0.0556
2
R = 0.9748
Samples Protease No protease
-1.2
Water
Buffered peptone water
Aqueous chicken extract
-1.6
0 10 20 30
Incubation time (min)

Fig. 3 Plot of cell population (Salmonella, 103 CFU/mL) growth rate under
different experimental conditions: (1) e Protease was added to aqueous chicken
homogenates at 0.5% (v/v), followed by inoculation of Salmonella; (2)
Salmonella was inoculated in aqueous chicken homogenates without addition
of protease; (3) Δ Protease was added to buffered peptone water at 0.5% (v/v),
followed by inoculation of Salmonella; (4) Salmonella was inoculated in
buffered peptone water without addition of protease; (5) □ Protease was
added to DI water at 0.5% (v/v), followed by inoculation of Salmonella; (6)
Salmonella was incubated only in the presence of DI water. Data are the
average of three assays. Error bars represent standard deviation. Population
growth rates up to 30 min of inoculation time are significantly different at the
95% confidence level. A similar pattern was observed when inoculating the cells
in chicken carcass rinses. (Vibbert et al., 2015; permission will be requested )

observed 80% viability is lost within 30 min when the cells are
suspended with protease in DI water. Therefore, a cell protec-
tive effect occurs when the protease and protein are incubated
together, keeping Salmonella cells viable [6–8].
6. For instance, one may be willing to concentrate and recover
cells originally present at very low levels (
1 CFU/g) in food.
In this case, a short enrichment step may be needed to increase
the cell number of the selected microorganism to detection
level after performing the cell concentration and recovery step.
7. We have tested a range of filters for the prefiltration step using a
vacuum pump (Fig. 4). The tested filters include: WhatmanVR
#4 (cellulose, 25 μm cutoff); Millipore Nylon net filter (nylon,
10 μm cutoff); WhatmanVRGF/D glass fiber filter membrane
(borosilicate glass, 2.7 m cutoff); and Pall Type A/E glass fiber
membrane (borosilicate glass, 1 mm cutoff). Scanning electron
 Accelerated Food Sample Preparation 15

(a) (b)

(e)

25 mm cut off 10 mm cut off

(c) (d)

2.7 mm cut off 1 mm cut off

Fig. 4 SEM (20 μm resolution) on the surface of (a) Whatman®#4 (cellulose, 25 μm cutoff), (b) Millipore nylon
net (nylon, 10 μm cutoff), (c) Whatman®GF/D glass fiber filter membrane (borosilicate glass, 2.7 μm cutoff)
and (d) Pall Type A/E glass fiber membrane (borosilicate glass, 1 μm cutoff) (e) Porcelain Buchner funnels with
fixed perforated plates for prefiltration (Adapted and modified from Ku et al., 2017, permission will be
requested )

micrographs of the four filters demonstrate that the filters with

the larger particle size cutoffs have faster filtration rates and
larger fiber diameters. In addition, higher pressure drops and
retention of smaller particles may lead to lower permeability as
result of the filter disk fouling. Particles that penetrated into the
filter not only restrict the passage of fluid, but also capture
micron-sized microorganisms, which may get retained in the
filter rather than exiting with the filtrate [8]. We intend with
this approach to remove 10–100 μm or larger particles that
block the 0.3–0.5 mm diameter hollow fiber, and allow micro-
organisms to pass in order to maximize microbial recovery. The
efficiency of a particular filter will depend on the processed
food type.
8. More often the enzyme treatment is performed before the
prefiltration step. However, for the ground Turkey samples,
the enzyme treatment followed by filtration caused submicron
particles to form and become trapped within the prefiltration
media, which in turn retained about 80% of the bacteria. On
the other hand, filtering prior to enzyme treatment resulted in
the formation of a filter cake of protein particles retained on the
16 Eduardo Ximenes et al.

Fig. 5 Automated continuous cell concentration device (C3D) in a duplex format.

The instrument is being upgraded to process 4–8 samples/run

surface of the filter, which facilitated the passage of the much

smaller microorganisms through the filter and separated them
from the particulates. The subsequent enzyme treatment of the
filtrate results in an extract that is microfiltered in less than an
hour and concentrated viable microorganisms in the
extract [8].
9. One possible trade-off of using glass microfibers is the loss of
microorganisms trapped in the membrane pores. This negative
effect is minimized by adjusting the protocol for a longer
enrichment step [8]. The GF/D glass membrane has also the
advantage of withstanding autoclaving, so that possible micro-
bial contaminants present in the filters are eliminated prior to
the experiments [3, 6, 8].
10. The schematic representation of the continuous cell concen-
tration device (C3D) is shown in Fig. 5. This research-based
prototype has been used to collect a dataset and optimize the
protocols described in this chapter (Fig. 1). A simplified ver-
sion of the instrument that can process multiple samples is
currently under development in our laboratory. The instru-
ment uses an on/off manual switch that activates a micropro-
cessor to direct the sequential cell concentration, recovery and
instrument cleaning.
11. For the research prototype, key microfluidic components con-
sist of a 4-to-1 source select valve (medium-pressure valve),
feed/retentate circulating pump (pump 1, compact analog
pump, 2 channels, 1.6 to 160 rpm, 0.004 to 50 mL/min,
115/230 VAC), membrane module, and a valve (three-way
valve 1/16 24VDC) that controls the direction of the retentate
flow. A second pump (pump 2, similar specifications as pump
 Accelerated Food Sample Preparation 17

1) pumps the sterile DI water into the membrane mode

through the permeate side to remove the generated permeate,
achieving uniform transmembrane pressure along the length of
the hollow fiber. The module’s cross-flow velocity is controlled
by adjusting the speed of pump 1. Teflon FEP tubing connects
the various components of the instrument. The flow rate of the
permeate (F1), the flow rate of the sterile DI water into the
permeate side (F2), the retentate pressure (P1), and the perme-
ate side pressure (P2) at the inlet end of the membrane module
are monitored using two differential pressure flow meters. The
operational control and data acquisition is conducted via
graphical interface in a program built with LabVIEW 2009
on a personal computer (PC) in our laboratory. The pressure
and flow rate data as well as the operation mode and the valve
statuses are automatically recorded each second. All operations
are carried out at room temperature. The instrument’s opera-
tion is defined by two operational modes—Cell concentration
and cell recovery. The microbiota of the prefiltered homoge-
nate are concentrated when the initial sample is fed through the
membrane module, being microorganisms concentrated in the
retentate, which is circulated into the sample reservoir using
pump 1.
12. In situations where selected microorganisms are expected to be
present at very low original levels (
1 CFU/g), a centrifuga-
tion step may be performed, after microfiltration for microbial
concentration and recovery, to further increase the microbial
cells number to detection levels (microcentrifugation at
3000  g and room temperature for 10 min). Also, our initial
results indicate that similar microbial concentrations may be
achieved with other Gram-positive (Escherichia coli) as well as
Gram-negative (Listeria monocytogenes) microorganisms.
13. Food homogenates are heterogeneous materials composed of
various components, including microorganisms, proteins,
lipids, micron-sized fine particles, and inorganic food compo-
nents. As we have mentioned previously, the proteins
contained in the liquid sample are known to accumulate on
the surface of the microfiltration membrane, forming a film
and/or blocking the filter pores, thereby impairing sample
filterability. Proteins are known to be a major fouling substance
of microfiltration membranes [6, 33, 34]. Regular chemical
cleaning is required to ensure reproducible performance and
repetitive membrane use, even if the tangential flow physically
removes the accumulated filter-cakes, reducing film formation
or deposition rate on the membrane surface [41].
14. The invA gene has been commonly used as the target sequence
for Salmonella species [43, 46].
18 Eduardo Ximenes et al.

15. When processing chicken rinses, PCR requires the use of a

DNA extraction/recovery kit because simple isolation using a
microwave to disrupt the cells and release DNA does not
remove the inhibitors. In this case, the use of a commercial
kit effectively removes such inhibitors [6].

Acknowledgments

The material in this work was supported by the FDA Food Safety
Challenge Prize and a cooperative agreement with the Agriculture
Research Service of the US Department of Agriculture project
(OSQR 935-42000-049-00D), the Center for Food Safety Engi-
neering at Purdue University, USDA Hatch project 10677, and the
Department of Agricultural and Biological Engineering at Purdue
University.

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 CHAPTER XV
Dance Tunes Grow Up—Suites—Violin Makers of Cremona

In our range of musical mountains, we see just ahead of us one of the

mightiest giants of them all, Johann Sebastian Bach, dwarfing
everything around it and we must resist the temptation of skipping
all the smaller mountains, for there is no musical aeroplane by
means of which we can fly across and land safely on Mt. Bach. This
grand old mountain, Bach, is such a tremendous landmark in the
growth of music, that when we reach it we realize that everything
that we have passed has been a journey of preparation. Bach is not
the only peak, for there are Mozart, Haydn, Beethoven, Chopin,
Schumann, Brahms, Wagner and others who stand out against the
musical horizon.
Before coming to Bach, however, we must bridge over the time
when music was still in its youth in the 16th and 17th centuries, to
when it became full grown and mature in the 18th. Music has now
come of age: it has perfected scales, notation, and developed form
and instruments; it is ready to go into the world and take its place
with painting, sculpture, poetry, drama and architecture as a full
grown art!
Nothing through which music has passed has been lost, but it has
been built like the great Egyptian Pyramids by adding one huge block
on top of another. It has gone from the noise of primitive man with
his drum, to the attempts of the savage to sing and to make crude
instruments, to the music of the ancient nations in their religious
ceremonies and entertainments, to the Arab singer who handed his
art to the western world through the troubadours, to the people of all
times and nations who danced and sang for the joy of it. It passed
from the Greek drama and music schools where definite scales and
modes were formed, to the early Christian Church which kept it alive
during the Dark Ages and gradually invented ways to write it, and
later to the “Golden Age” of the Catholic Church. It had seen the rise
of schools and the perfection of the polyphonic system give way to
the recitative and the aria, which in turn brought about opera,
oratorio, and instrumental music. It has seen counterpoint give way
to harmony, and yet the growth of music is not complete and never
will be, but constantly new forms will blossom out of the old.
The 15th and 16th centuries were vocal. The 17th was instrumental
and opened the way for so-called modern music, that is, for Bach’s
compositions and all that followed.
 Birth of Chamber Music

Gabrieli in the 16th century in Venice, sometimes wrote madrigals

for instruments instead of for voices, and he added instruments to
accompany the motets and masses (page 157); this led to composing
works for groups of instruments instead of playing madrigals that
had been composed for voices. The English often wrote on their
compositions, “fit for voices and for viols.” After they once started
playing the part songs on viols, the composers soon found out that
they could write more interesting and more difficult things for
instruments than they could for voices; this led to the writing of very
florid music for instruments alone. This florid part-writing, not
unlike the Gloss of the Arabs, and the improvisations of the soloists
in the early Catholic Church, soon became so overloaded with trills,
fancy turns and runs that it had to be reformed again.
In the 17th century, the lute, the popular instrument of the court
and the home for so many years, even centuries, suddenly found its
rival in the little keyboard instrument called the spinet and virginal
in England, and the clavecin in France. In Italy and France, as in
England, there were famous performers and composers for these
instruments, and many volumes of charming music were written for
it.
 Dance Tunes Grow up Into Suites

One of the first requirements of art works of all kinds is contrast.

The line and the curve are found in primitive art, light must have
shadow, one wing of a building must have another to balance it, and
a slow serious piece of music is usually followed by a gay one for
contrast. The Arabs understood this law of contrast, for in their
ancient songs we find the seed of a form that has been most
important in the growth of music. They made little suites by putting
two, three, four or more songs together; each song had its mode, and
one would be slow and sad, and the next fast and gay. The principal
music of the 17th century was the Suite, a group of pieces which had
grown out of the old folk dances. (Chapter IX.) The 17th century
composers, like the Arabs, feeling the need of contrast, strung several
of these dances together to form the Suite. So Suites were written for
clavecins and harpsichords, for violins alone and for organs, for
groups of stringed instruments and other chamber music
combinations. Some of these dances were in duple time, some in
triple; some were slow and some were fast; some were stately and
some gay. The different pieces forming a suite, had to be written in
the same key. These suites were known by different names in
different countries, such as partitas, exercises, lessons, sonate da
camera, ordres. In England the name suite was given to this form,
then the Germans adopted it, and later the great Bach wrote suites
which he also called partitas. In Italy, the suite was called sonata da
camera (chamber sonata) and sonata da chiesa (church sonata) and
out of all this have grown the very important sonata, symphony and
chamber music quartet, trio, quintet, etc.
Here are some of the dance forms used in the suite:
Allemande (duple time or measure: moderately slow), Sarabande
(triple time: slow, stately), Loure (duple time: slow), Gavotte (duple
time: moderately fast), Musette (duple time: moderately fast),
Bourrée (duple time: a little faster than the Gavotte), Minuet (triple
time: moderately fast), Passepied (triple time: a fast minuet),
Rigaudon (duple time: slower than the Bourrée), Tambourin (duple
time: fast), Pavan (duple time: rather slow), Courante, Corrente
(triple time: fast), Chaconne (triple time: moderately fast),
Passacaglia (like Chaconne, but more stately) and Gigue (sometimes
duple and sometimes triple time: very fast: almost always the last
movement of a suite).
The Italians of the 17th century wrote suites, and Italy still held the
place as leading the world in musical composition, just as it had in
the 15th and 16th. We find the names of Frescobaldi, Michelangelo
Rossi, Legrenzi, Bononcini, Giovanni Battista Vitali, Alessandro
Scarlatti and his son Domenico, and going over into the seventeen-
hundreds, Niccolo Porpora, Padre Martini, Paradies, and Baldassare
Galuppi, whom we know through Robert Browning’s poem, A
Toccata of Galuppi’s. Most of these names you will find on the
concert programs of today.
 “Serious” Scarlatti and Opera Writers

Alessandro Scarlatti (1659–1725) is one of the most important

Italian composers of the 17th century, and although he did not have
great success during his lifetime, his compositions have outlived
those of other writers, whose works were popular during his day. He
was called “serious Scarlatti,” and it was probably the very
seriousness with which he looked upon his work that made him write
without seeking public approval. Besides composing pieces for the
spinet and harpsichord, and symphonies, sonatas, suites and
concertos for different instruments, he wrote 125 operas, and over
500 cantatas, oratorios and church music. He was one of several
Italians who continued the work of the first opera writers. Francesco
Cavalli (1599–1676), Giacomo Carissimi (1603–1674), Luigi Rossi,
Marc Antonio Cesti (1628–1669), Francesco Provenzale (1610–
1704), Stradella (1645–1682), Caldara (1670–1736), Lotti (1667–
1740), Marcello (1686–1739), Leo (1694–1746), and others carried
the ideas of Scarlatti into the 18th century. Many of these carried
Italian opera into England, Germany and France, where it became
the model for their opera.
Stradella is quite as famous for his romantic love story, as he is for
the operas he left. This made an interesting libretto in the 19th
century for a German opera writer, Flotow, who was also the
composer of the well-known opera Martha.
 “La Serva Padrona” Points the Way

Giovanni Battista Pergolesi (1710–1736), who died when he was

only twenty-six years old, was looked upon as a genius, and in his
early youth had written two works that were models for many that
followed, a Stabat Mater and a comic opera, La Serva Padrona,
which was played recently in America under the title of The Mistress
Maid. When this little opera was performed in Paris (1752) it caused
a very famous musical quarrel known as the “war of the buffoons.”
(Page 230.)
Jomelli (1714–1774), the composer of fifty-five operas, was a
Neapolitan but he lived in Germany for so many years, that he had
more influence on early German opera than on the Italian.
All the opera of this period, particularly the Italian, was very
loosely put together and was not opera as we have it today. Later
Gluck brought it to the point where it came of age.
 Metastasio—Maker of Opera Librettos

These writers of the 18th century used the librettos of a poet and
dramatist, Metastasio (1698–1782), who had a strong influence in
the development of opera not only in his native Italy, but in other
countries. He supplied texts for 1200 operatic scores! He understood
music so well, that he was a great help to the composers who listened
with attention to his advice. His life covered practically all of the 18th
century.
 A Celebrated Singing Teacher and Composer

When you read of Haydn, you will see that he played

accompaniments and acted as valet to the eminent singing teacher
Niccolo Porpora (1686–1767). This famous Italian had many pupils
in the opera houses all over Europe, and was considered the greatest
singing teacher in the world. One of his pupils in Dresden was the
young princess Marie Antoinette before she became Queen of
France. Porpora was a fine composer, and wrote many operas,
cantatas, masses, oratorios, and sonatas of which form he was one of
the inventors. Among his pupils were Haydn, Marcello, Tartini, Leo,
Galuppi, Padre Martini, Jomelli, Pergolesi, Caffarelli and Farinelli.
This list shows that he trained composers as well as singers.
 The Violin Makers of Cremona

Important changes, such as instrumental music coming into

fashion, do not happen without good reasons. We are so accustomed
to the violin, that we forget that there was a time when it did not
exist, but until about three centuries ago, there was none. We are
always eager to have new pianos, for the old ones wear out, but with
violins the older they are, the better! But they must be masterpieces
to begin with. All the famous violinists of the day like Kreisler,
Elman, Heifetz, etc., have marvelous old violins that cost fortunes,
and most of them were made by the violin makers of Cremona, a
little town in northern Italy, the birthplace of Monteverde.
The troubadours played the accompaniments to their songs on
stringed instruments called violes or vielles, which were the
grandparents of the violins. In the 15th century bowed instruments
were made similar in range to the human voice; these were called
treble or discant viol, tenor viol, bass viol and the double-bass, and
in England these went into the “chest of viols” (Page 198). Many
improvements were made in the shape, size and tone of the
instruments and by the middle of the 17th century the Italian makers
were ready to create violins, perfect of their kind, which have never
been surpassed. The secret of the tone of these instruments is said to
be in the varnish which the Cremona makers used, the recipe of
which has been lost, but we met a violin maker recently in Paris who
had discovered it in an old Italian book, and he has spent years in
trying to reproduce it. The old Italian varnish and the mellowing of
the wood with time are two reasons why age makes the old violins
better.
For several centuries, practically all the lutes and the viols that
supplied Europe were made by colonies of instrument makers who
lived in Lombardy (North Italy) and the Tyrol (South Austria). Two
towns in Lombardy became especially famous for their violins,
Brescia in which Gaspara di Salo and Maggini lived, and Cremona
which was the home of the Amati, Stradivari and Guarneri families.
In The Orchestra and Its Instruments, Esther Singleton says: “It is
thrilling to realize that in this little town, in three workshops side by
side, on the Piazza San Domenico, all the great violins of the world
were made and in friendly competition by the three families.” This
covered the period from 1560 to 1760. These men worked together
with just one object in life,—to turn out of their shops the most
perfect instruments that could possibly be made! With what care
they selected the wood! How they worked to make the tone of each
instrument as beautiful as possible! Now you will know when you
hear of an Amati violin, or a Stradivarius, a Guanerius or a Maggini,
that they are worth their weight in gold and are among the rarest art
treasures of the world. These were not the only violin makers in
Lombardy, for there were long lists of them, and there were also
many in the Tyrol. One of the most famous of these was Stainer who
lived at Innsbruck. “It is said that this old maker used to walk
through the wooded slopes of the Tyrolean mountains with a
hammer in his hand and that he would knock the trunks of the trees
and listen to the vibrations. When he found a tree that suited him, he
had it cut down to use in making his instruments.” (Esther
Singleton.)
These instrument makers made not only violins, but also lutes,
mandolins, guitars, violas, violoncellos, and double-basses. The
Italians were the first to develop the last two. The ’cello, as we call
the violoncello for short, was the child of an instrument named the
viola da gamba (translated leg-viola because it was held against the
leg), which for many years was the most popular of all bowed
instruments. We do not find many examples of the instruments even
in museums for they were made over into ’cellos when the latter
came into fashion. There is one viola da gamba in the Metropolitan
Museum of Art, however, which was imported from France by the
Sisterhood of the General Hospital in Montreal before the conquest
of Canada, and was used in the convent choir many years before
there were any organs and pianos in the New World. The first ’cello
to attract attention was made in 1691 by a famous wood carver and
presented to the Duke of Modena. A member of the Amati family in
the 16th century was the first to turn the viola da gamba into a
violoncello. The ’cello and the double-bass were made more
successfully by Bergonzi than by the Cremona makers, although
Maggini, Amati and Galiano made very fine ones.
 The viola is a descendant of the viola d’amore. These and the later
violas, used in the string quartets, orchestras, and as solo
instruments, were made by a Tyrolese named Gaspard Duiffaprugcar
in the 16th century. His instruments are marvelous works of art. In
the back of one is a riddle in Latin: can you guess the answer? “I was
living in the forest; the cruel axe killed me. Living, I was mute; dead,
I sing sweetly.” When madrigals and motets were first played on
stringed instruments, the principal melody was given to the tenor
viol, the ancestor of the viola, even today called the alto or the tenor,
but after the violin came into general use, the viola was treated like a
step-child, for it is too large for a violin and too small for a
violoncello. We have Mozart to thank for discovering that the viola
had something beautiful and important to say as a solo instrument
especially in passages where he needed a tender, sad or melancholy
voice. You will read later that Beethoven, too, loved the poor
neglected viola. He, Berlioz and Wagner used the instrument to great
advantage.
In 1572 Pope Pius V sent Charles IX, King of France, a present of
thirty-eight bowed instruments made by the first Amati. During the
French Revolution, the mob broke into the palace at Versailles, and
all but two violins and a ’cello were destroyed! What a loss to art such
destruction was!
 Showing Off the New Instruments

With this development of exquisite instruments, came the desire to

use them and to write new compositions to show them off. These
instruments gave unlimited possibilities for technic and tone, and
created the school of Italian violinists and composers of the 17th and
18th centuries. If polyphonic music had still been in the lead, the
development of solo instruments would have been impossible, but in
trying to find new forms, the first opera inventors had broken the
backbone of polyphony, and had replaced it with monody, or single
line melody. Then, too, folk dances had taken the public fancy and
had been made into suites, which could be played on solo bowed
instruments with accompaniments, on spinets and organs, or on
groups of instruments. The sonata da camera was really a suite of
dances and was the first form used by these new composers for
violin. About the middle of the 17th century, instrumental
performances without any vocal music came to be a part of the
services of the Catholic Church for the priests were quick to see in
the violin playing, a refining influence. Here the sonata da camera
or “room sonata” was turned into the more serious sonata da chiesa
or “church sonata” gradually losing its dance character, and thus
became the seed of the sonata form of Haydn, Mozart and
Beethoven.
Giovanni Battista Vitali (1644–1692) is the first great master of the
violin sonata; after him, Torelli (1657–1716) added a new and
important kind of violin composition,—the Concerto. He called his
compositions, Concerti da Camera and Concerti Grossi, which
names and form were used by Vivaldi, Corelli, Handel and Bach. This
Concerto Grosso was a sonata da chiesa accompanied not by a single
instrument as was the habit with the sonata da chiesa and the
sonata da camera, but by a group of bowed instruments to which a
lute, organ and, later, a harpsichord were added.
At this time, all musicians were, as a matter of course, violinists,
just as today all great composers can play the piano. One of the
greatest of these composer-violinists was Arcangelo Corelli (1653–
1713), whose works are often played by violinists of our own time,
and have served as models for composers. He was one of the first to
try to write music that should show off the beauty and possibilities of
the violin.
The “Golden Age” of the Italian violin composers dated from 1720
to 1750, and was the time of Locatelli, Pugnani, Nardini, Veracini,
Tartini and Vivaldi who added oboes and horns to the orchestral
accompaniment of the Concerti Grossi. Corelli and Vivaldi were the
models used by the German school of violinists who appeared about
this time. Tartini was the musical authority of his century, and no
violinist felt sure of his place as an artist until he had been heard and
approved by Tartini. He was the composer of the famous piece called
The Devil’s Trill. Although Vivaldi was not looked upon with great
esteem in his own time, he was used as a model by Johann Sebastian
Bach.
Padre Martini, recognized by all Europe as the greatest authority
on musical subjects, lived in Bologna where he was visited by such
musicians as Grétry, Gluck, Mozart and one of the sons of Bach.
Padre, or Father, Martini was a Franciscan monk, a fine composer, a
learned historian, a master of counterpoint, and the owner of a
musical library of 17,000 volumes! He helped everyone who sought
him, and was loved by the entire musical world.
Once a year a great music festival was held in Bologna by the
Philharmonic Society and new works by the Bolognese composers
were performed. One hundred musicians took part in the orchestra
and the choruses, and each composer conducted his own work. It
was an honor to be present at this annual festival, and Italian and
foreign musicians came from all over Europe to attend it. Young
composers sometimes became famous over night here, for the critics
were all invited and serious decisions were made as to the value of
new music. Dr. Burney, a famous English musical historian of the
18th century, tells of meeting Leopold Mozart and his young son,
Wolfgang Amadeus, at one of these festivals. Through the kind
scheming of Padre Martini were they admitted!
Rome, in the 18th century was still the great music center, and
guided the religious music of the world. It had wonderful collections
of old music which attracted students from all over; it had seven or
eight very famous theatres, where opera seria and opera buffa were
given. (Today we call them grand opera and comic opera.) The
Roman public was very difficult to please and because of the severity
of their judgments, opera writers suffered every time their new works
had first performances. Just think how you would feel if you had
composed an opera, and by accident had put in a melody that
sounded something like one that Mozart, Wagner, Puccini or Verdi
had composed, if the whole house should break into shouts of
“Bravo, Mozart!” or “Bravo, Wagner!” or “Bravo, Puccini!!” etc. This
is what used to happen in Rome, but no doubt it was a good thing
because it stopped a habit the composers had in those days, of
helping themselves to each other’s melodies.
 Domenico Scarlatti

But here we must pause for a moment to tell you of the life and
work of Alessandro Scarlatti’s son, Domenico, who was born in
Naples in 1685, the same year as Bach and Handel. When you recall
how many operas the father wrote, it seems queer that his son did
not follow in his footsteps. The truth is that he did write operas for
the private theatre of the Queen of Poland in Rome, and also sacred
music while he was chapel master of St. Peter’s, but he became
immortal as a composer of harpsichord music. In the influence he
had in the growing up of piano music, he can be compared to Chopin
and Liszt, and is a founder of piano music style, an honor, which he
shares with the French Couperin and Rameau, his contemporaries.
The difference is that the two Frenchmen have a delicacy and grace
that recall their period of wigs and satins and laces, while Scarlatti’s
works have strength, vigor and daring that take them out of any
special period and place them beside the great piano compositions of
all time.
Scarlatti’s sonatas are sonatas in the Italian sense of a sound-piece;
they are not, like the suites, in several movements, but each is in one
movement, which forecasts the modern sonata form with its two
main contrasting themes and development.
The “serious Scarlatti” understood his son’s talent, for he sent him
at the age of 20 to Florence to a member of the powerful de Medici
family with this letter: “This son of mine is an eagle whose wings are
grown; he ought not to stay idle in the nest, and I ought not to hinder
his flight.”
Three years later Handel and Scarlatti met in Rome in an organ
and harpsichord competition, and while Handel won as organist,
even Scarlatti declaring that he did not know that such playing
existed, no decision was made as to which was the better harpsichord
player. This contest seems to have caused no hard feelings for the
two young men of the same age became devoted friends.
Scarlatti had a trick of crossing his hands in his compositions.
Who does not remember with joy his first piece in which he had to
cross his hands? But sad to relate as he grew old, he became so fat
that he could no longer cross hands with comfort, so in the last
compositions the crossing of hands is noticeably absent!
It is hard to know where an inspiration is next coming from, but
wouldn’t you be surprised were you a composer, if your pet cat
presented you with a perfectly good theme? This happened to
Domenico Scarlatti! His cat walked across the keyboard, and the
composer used his musical foot prints as the subject of a very fine
fugue! Maybe Zez Confrey’s Kitten on the Keys is a descendant of this
pussy’s piece.
The Scarlattis were the last of the great Italian instrumental
composers. For two centuries Italy had been the generous dispenser
of culture, and like an unselfish mother had sent her children out
into the world to carry knowledge and works to all the nations of
Europe. The sun of Italy’s greatness was setting just as it began to
rise in Germany.
 CHAPTER XVI
Opera in France—Lully and Rameau—Clavecin and Harpsichord
Composers

We left French Opera in 1600 when Henry IV married Marie de’

Medici. Ballets which resembled the English masques had been
performed when Baif and his friends had produced Le Ballet
Comique de la Reine, but no real opera had yet been written in
France. In 1645, Cardinal Mazarin, the powerful Italian prime
minister of France, invited a company of Italian singers to give a
performance of Peri’s Euridice in Paris. The French did not like the
opera, as they said it sounded too much like plain song and airs from
the cloister, and yet it led to Abbé Perrin’s writing a work in 1658
which he called the Pastoral, and for which a composer named
Cambert wrote the music. The Pastoral was a very great success, and
was repeated by order of Louis XIV, King of France. Ten years later,
Louis gave Perrin and Cambert permission “to establish throughout
the kingdom academies of opera, or representations with music in
the French language after the manner of those in Italy.” Their next
work, Pomone, was the first opera performed publicly in an opera
house, built purposely in Paris for them. The opera was so
enthusiastically received, that it ran nightly for eight months, and the
crowds were so great, that the police had to be called out. This
combination of poet and composer came to an end with Pomone, and
a new man acquired the right to give opera in the new opera house.
This man was Jean Baptiste Lully or in Italian, Giovanni Battista
Lulli (1632–1687).
 Lully the King’s Favorite

You may hear that the first famous opera writer of France had
been a pastry cook or kitchen boy, but no matter how humble his
start in life, he rose to the highest social position ever reached up to
that time by a composer in France. He became a great favorite of
Louis XIV, he was covered with titles and honors, he was on friendly
terms with all the nobility of the court, he was musical dictator of the
opera and in fact of all the musical happenings of the court. The
greatest literary geniuses of the period, such as Molière, Racine, La
Fontaine, Quinault, Corneille and Boileau, worked with him when he
wanted new librettos for his operas. He paid dearly for all his
privileges, because his fellow composers were jealous of his genius
and his opportunities, and they lost no chance to blacken his
character.
Lully was born in Florence, Italy, in 1632, but we can tell you little
or nothing of his parentage or of his childhood. A monk taught him a
little about music and how to play the guitar. When he was about
twelve years old, he was picked up by the Duke de Guise who saw
him with a group of traveling comedians, and was so attracted by his
vivacity, his singing and talent for mimicry, that he took him back to
Paris, where he placed him in the household of his cousin, Mlle. de
Montpensier. In her memoirs, Mademoiselle said that she had been
studying Italian and had asked her cousin to bring back from
Tuscany where he lived, a little Italian garçon de la Chambre, a sort
of personal errand boy. However, his guitar playing and musical gifts
soon lifted him out of a servant’s position and he became one of the
musicians of the great lady’s household playing at concerts, balls and
in the ballets. He learned to play the violin, and soon began to
compose popular dances. He remained a member of Mademoiselle’s
household until he was nineteen when he asked permission to leave
her service, as she had moved to the country, and he liked the gay life
of Paris better.
He had no difficulty in attaching himself to the King’s court, first
as actor and dancer in the ballets, and soon as “composer of
instrumental music.” Louis XIV was only fourteen years old, and was
evidently highly entertained by the capers of the young Italian who
was willing to play any rôle, dance any kind of a dance, or play the
violin “divinely” for his young monarch’s amusement. The King
remained Lully’s faithful friend always. Louis loved music, and
played the lute, the guitar, the harpsichord, and sang very well.
Feeling that he needed to know more, Lully studied counterpoint,
composition and learned to play the harpsichord, and whatever he
attempted musically, he acquired without difficulty.
In 1656, Lully composed music for a scene in a ballet, Psyche, and
from that time on, his compositions became the most popular of any
at court. Although he was born an Italian, his music was French, and
he even shared the French dislike of the Italian opera. In spite of his
love of acting in the ballets, of dancing, and of courting social favor
with the King and nobles, Lully was a thorough musician. When he
went into music he found that few of the singers could read notes,
but they learned their parts by ear. He soon changed this, and by the
time he died, all singers and players of orchestral instruments could
read well. In this reform, he did a great service to the growth of
music.
His first stage works were called comedy-ballets. One of his early
works was ballet music written for a performance of Cavalli’s opera,
Xerxes, which was performed upon Mazarin’s invitation at Versailles
(1660). He next was given the position of “Superintendent of Music,”
became a naturalized French citizen, and was married. Lully wrote 19
ballets, 12 comedy-ballets, and 18 operas, besides about 23 motets
for special occasions. His ballets included recitatives, airs, dialogues
and symphonies, which was the name given to music written for
orchestra. From 1672 until the time of his death in 1687, he wrote an
opera a year, and sometimes two!
The splendor and extravagance of the costuming and stage settings
of these ballets and operas of Lully are almost unbelievable! At times,
even the orchestra wore costumes of the period represented on the
stage. Lully conducted the orchestra for one opera in a magnificent
Egyptian dress. Louis XIV loved these elaborate performances, and
took part in some of them.
After the downfall of Perrin and Cambert, which many said was
caused by Lully, he became absolute ruler in all musical matters. He
used his power to close a rival opera house, and no opera could be