NAME :-Joytri Saha

CLASS :-XI
SEC :-Pine
ROLL :-10
SUBJECT :-Chemistry
TOPIC :-Study of the acidity of various fruits and
vegetable juice
ACADEMIC SESSION :-2022-2023
1|Page

PREFACE
As a student of class 11th Science and in order to gain
practical knowledge in Chemistry, we require to make
a project copy on any of provided topic choices.
This file includes a detailed theory on the study of
acidity of various fruit and vegetable juices.
2|Page

ACKNOWLEDGEMENT

I would like to express my sincere gratitude to

my chemistry teacher Mr. Soudam Sir for his vital
support, guidance and encouragement without
which this project would not have come forth. I
would also like to express my gratitude to the lab
assistant for his vital support during making of this
project I wish to thank my family who contributed
in one way or the other and supported morally
and financially towards the completion of this
project.

JOYTRI SAHA
ROLL NO.-10
CLASS-11(PINE,SCIENCE)
3|Page

CONTENT
INDEX PAGE NO.
1. Preface 01
2. Content 02
3. Acknowledgement 03
4. Aim 04
5. Introduction 05
6. About the project 05-17
7. Conclusion 18
8. Reference 19-23
4|Page

AIM
TO STUDY OF THE ACIDITY OF VARIOUS FRUITS
AND VEGETABLE JUICE
5|Page

INTRODUCTION
Juice is a liquid naturally contained in fruit or vegetable tissue. Juice is prepared by
mechanically squeezing or macerating fresh fruits or vegetables without the application of heat
or solvents. For example orange juice is a liquid extract of one fruit of orange tree. Juice may be
preparedin the home from fresh fruits and vegetables using variety of hand or juice extractor.
Many commercial juices are filtered to remove fibre or pulp, but high pulp fresh orange juice is a
popular beverage. Juice may be marketed in concentrated form, sometimes frozen, requiring the
user to add water to reconstitute the liquid back to its original state. However, concentrates
generally have a noticeably different taste than their comparable fresh squeezed versions. Other
juices are reconstituted before packaging for retail sale. Common methods for preservation and
processing of fruit juices include canning, pasteurization, freezing, evaporation and spray drying
(Fasoyiro et al., 2005) . However, fruit juices are nutritious which offer great taste and health
benefits. The 2005 Dietary Guide lines for Americans (2005) recommended consumption of
several cups Effect of keeping time period on acidity of fruit juices and determination of fungal
growth in fruit juices per day of fruits and vegetable. Most fruit juices bought from grocery
stores and supermarket shelves are pasteurized. This means that the liquid has been brought to
a high temperature that kills harmful bacteria. However, a small percentage of fresh fruit juices
are unpasteurized. This means that there is a chance that the product may contain bacteria
harmful to our health. Most people can enjoy unpasteurized juice and drinks, however, for
young children, the elderly and people with weakened immune systems, the effect can be severe
or ever deadly (Fasoyiro et al., 2005).Unpasteurized fruit and vegetable juices have posed
serious public health risk in recent years. Seventy people including a child who died-became ill
in 1996 after drinking unpasteurized apple juice contaminated by a strain of Escherichia coli
bacterium ( Amato,1999). Fruit juices are well recognized for their nutritive value, mineral and
vitamin content. They are beverages that are consumed for their nutritional value, thirst-
quenching properties and stimulating effect or for their medicinal values(Fawole and Osho,
2002). The low pH of fruit juices greatly limits the number and the type of bacteria that can
survive or grow at this low pH but some bacteria that their pH is lower than that of the fruit
juice can grow at this condition (Ryu and Beuchat, 1998). Yeast and moulds are also present and
can grow when the juice is held at a temperature permitting their growth.Yeasts are primarily
responsible for the spoilage of chilled juice that is not sterile and some can withstand the effect
of chemicals used to preserve them (Sandeep et al., 2001).In India, there is always a great
demand for fresh vegetables and fruit juices. Being tropical in location hot weather continues
for a greater part of the year (February-September) increasing the need for these commodities.
While most restaurants and cafes serve juices in apparently hygienic conditions in the roadside
shops and recreational areas and busy market places, their microbiological quality remains
questionable (Sandeep et al., 2001). In these shops juices extracted by squeezing from a variety
of fresh fruits namely,oranges, grape, pomegranate, apple, pineapple, watermelon,papaya,
carrot and soon were served after considerable dilution with water and ice
(Splittstosser,1979).In views of high demand of fresh fruit juices during different diseases, fruit
juices was under taken with a view to assess their safety for human consumption (Uljas and
Ingham,1998). This study was therefore aimed at : to study the pH of fruit juices, acidity of fruit
juices, the effect of keeping period on acidity of fruit juices, and determine the fungal growth in
fruit juices. Over the years, three groups (aciduric bacteria, molds, andyeasts) have been
reported as the most important micro organisms since they are acid tolerant. In the past 30
years, however, the emergence of a sporulating acido-thermophilic bacterium, Alicyclobacillus
spp. (see Alicyclobacillus), has been observed. Yeasts, heat-sensitive molds, and lactic acid
bacteria (LAB) are important indices for the quality of raw materials. Heat-resistant fungi and
other sporeformer bacteria, such as Clostridium pasteurianum (see Clostridium) or Bacillus
6|Page

coagulans (see Bacillus: Introduction) and Alicyclobacillus acidoterrestris play an important

role as targets for fruit juice pasteurization processes, depending on the time and temperature
conditions employed (e.g., mild treatments). Until the 1980s, it was believed that under acidic
pH values (pH <4.5), pathogen growth would not be observed, while survival, although possible,
would be improbable. Foodborne disease outbreak occurrences, however, resulted in more
attention being given to acidic fruit juices, for example, apple cider, which has been implicated
in hemorrhagic colitis caused by Escherichia coli O157:H7. Recently, the rapid dissemination
and search for exotic juices with high pH, such as melon (5.5–6.0) and watermelon (5.2–6.8),
has brought a new challenge to the fruit juice industry. This challenge is related to the fact that
these juices provide good conditions not only for the survival but also for the growth of
foodborne pathogens.

Lactic Acid Bacteria

The genera of Lactobacillus (see Lactobacillus: Introduction) and Leuconostoc (see The
Leuconostocaceae Family) are the most important groups of spoilage microorganisms for acid
products. The heat-resistance index (D-value) for Lactobacillus spp. In fruit juices at pH 4.0 and
53 C is 0.6–1.9 min and for Leuconostoc mesenteroides, 1.5 min at the same conditions. A thermal
treatment of 95 C 30 s1 was able to totally destroy a 106 log cfu ml1 load of LAB existing in orange
juice. A mean LAB contamination level was reported for raw juice before pasteurization of 1.4E þ
04, 3.4E þ 05, and 2.0E þ 6 cfu ml1 for single-strength orange juice, apple nectar, and grape
beverage, respectively. The evolution of the LAB contamination in two orange juice–processing
plants is shown on Figure 1. Contamination before pasteurization can reach 107 cfu ml1, but it is
totally eliminated by pasteurization. Although the presence of LAB is more commonly reported
in unpasteurized juices, contamination after the thermal processing of fruit juices plays an
important role. Spoilage episodes involving these microorganisms are the result of failures in
cleaning and sanitation programs, mainly from equipment that comes after the pasteurizer on the
production line. The process line contamination by LAB may result in biofilms formation
(dextran), which can increase LAB heat resistance.
7|Page

Acetic Bacteria
Acetic bacteria have been associated frequently with beverage’s spoilage, characterized by
changes of flavor, viscosity, and gas formation. The increased use of plastic packages leads to a
higher incidence of nonfermentative acetic bacteria, mainly Gluconobacter (see Gluconobacter),
which are resistant to chemical agents and able to grow in presence of higher concentrations of
oxygen and sorbic and benzoic acids.

Yeasts
Yeasts are able to grow under low pH conditions, high sugar content, and refrigeration
temperatures, making them potential spoilers of simple and concentrated juices (see Yeasts:
Production and Commercial Uses). Their growth spoils the juices by producing carbon dioxide
and alcohol, enhancing turbidity, and causing flocculation and phase separation due to the action
of microbial enzymes on the pectin. Candida (see Candida), Pichia, Rhodotorula (see
Rhodotorula), Torulopsis, Saccharomyces (see Saccharomyces – Introduction and
Saccharomyces: Saccharomyces Cerevisiae), Zygosaccharomyces (see Zygosaccharomyces),
Hansenula, and Trichosporon genera have been associated with juice spoilage. For citrus juices,
Candida stellata, Saccharomyces cerevisae, Torulaspora delbrueckii, and Zygosaccharomyces
rouxii have been reported. Since these organisms tolerate high-osmotic pressure, low pH
conditions, and grow at refrigeration temperature, they can cause spoilage in the processed
products. The heat resistance of yeasts representative of the fungal flora of soft drinks, and certain
acid products has been investigated. Generally, asporogenous yeast strains have been found to be
less heat resistant than ascomycetes (see Fungi: Classification of theEukaryotic Ascomycetes and
Fungi: Classification of the Hemiascomycetes). The genus Saccharomyces showed the highest
heat resistance, particularly strains of species S. cerevisae.

Molds
Molds are mostly aerobic, tolerate low pH values, and tolerate high sugar concentrations. They
can produce gas, change the odor, and form mycelial mats on the juice surface. Most fungi show
limited heat resistance. The asexual spores (conidia) of the very common genera, such as
Penicillium (see Penicillium and Talaromyces: Introduction and Penicillium/Penicillia in Food
Production), Aspergillus (see Aspergillus), Mucor (see Mucor), Rhizopus, Fusarium (see
Fusarium), Cladosporium, and Botrytis are killed after heating for 5 min 60 C1. The vegetative
cells of these genera are also inactivated within 5–10 min if heated at 60 C in distilled water. Some
more heat-resistant species owe their resistance to sclerotia or to thick-walled sexual spores
(ascospores); sclerotia of Penicillium, which causes spoilage of canned blueberries, survived
heating at 85 C 4.5 min1. The most important molds in fruit juice industries can be divided
considering their response to thermal treatment as heat-sensitive (labile) and heat-resistant
molds (HRMs). Inactivation of Alicyclobacillus niger IOC 4573 conidia in mango nectar (pH 4.05,
14 Brix) at 80 C has been evaluated.It was verified that this strain was able to survive a heat shock
of 80 C 30 min1, which characterize HRMs. Although heat resistance is not common for
Aspergillus species, some researchers have isolated strains able to survive 85 C 60 min1 in grape,
apple, and tomato juices.

Heat-Resistant Molds

Spoilage due to formation of heat-resistant ascospores by members of the genus Byssochlamys

(see Byssochlamys), Neosartorya, Talaromyces, and Eupenicillium has been reported repeatedly.
Some strains of Byssochlamys and Neosartorya fischeri have become an industrial problem by
spoiling processed fruit products and producing mycotoxins: byssochlamic acid, patulin, and
8|Page

byssotoxin A (see Mycotoxins: ClassificationNatural Occurrence of Mycotoxins in Food,

Mycotoxins: Detection and Analysis by Classical Techniques, Mycotoxins: Immunological
Techniques for Detection and Analysis, and Mycotoxins: Toxicology). Patulin may be produced by
species of Aspergillus,Penicillium, and Paecilomyces but mainly by Penicillium expansum.Patulin
has been found in apples, pears, their juices and jams; grapes and grape juices; and beet juice. It
was also found in fruits that exhibited brown rot, such as bananas and pineapples. Byssochlamys
species are historically among the most widely encountered molds causing spoilage of heat-
processed fruits and therefore have been studied extensively. This genus can survive the thermal
process given to many acid foods and have been responsible for spoilage outbreaks of
commercially canned fruits and fruit products. In the 1990s, however, spoilage due to N.fischeri
and Talaromyces fravus had been observed more frequently in North America, Europe, and
Australia. In Table 1, we have summarized more recent studies conducted in juices, nectars, and
pulps, involving such genus as Neosartorya, Talaromyces, and Byssochlamys. Neosartorya
fischeri is by far the most resistant, and in almost all cases, the inactivation is nonlinear. The heat
resistance was compared between ascospores of Byssochlamys fulva and Aspergillus spp. WR1
in 5 Brix Concord grape juice, pH 3.5 were compared. Similarities in behavior included a
nonlogarithmic order death, and increased heat resistance in sugar solutions were observed.
9|Page

Incidence of HRM

The preservation of fruit and vegetable juices is based mainly in the heat process. The addition of
chemical or natural antimicrobials to fruit juices to avoid microbial growth during the shelf life
has also been well studied. Consumer demand for natural products presents a big challenge to
juice industries,however, attempting to ensure microbiologically stable products without the
addition of reservatives.In Brazil, 58 strains of HRM were isolated from strawberry pulp.
Byssochlamys nivea was the most heat-resistant strain tested, and it showed a nonlinear behavior
with survivor curve with shoulder and tail. Similar behavior has been reported in other countries
located in the Southern Hemisphere, such as Argentina, New Zealand, Australia, and South Africa.
The occurrence of HRM in raw material and the subsequent increase in number of these species
in Nigerian final products were also studied. The number of ascospores in fruits is in general low,
less than one per gram. Approximately 98% of all soil samples and 17% of mango fruits samples
contained HRM, which were identified as N. fischeri var. spinosa, Aspergillus flavus, Penicillium
citrinum, and Paecilomyces variotii. Neosartorya fischeri was predominant and occurred in all
positive samples. This fact is very important since certain strains of N. fischeri are capable of
producing mycotoxins, such as fumitremorgins A, B, and C, and verruculogen. These compounds
have been proved to act on the central nervous system. The occurrence and the heat resistance
of HRM during the aseptic processing of Brazilian tomato pulp (8 Brix) were studied. The higher
counts were observed in the raw material, prewash, and transportation water. The reutilization
of condensate water for fruit washing may lead to an increase in the contamination. Fifty strains
of HRM were isolated and the most heat-resistant strain was identified as N. fischeri and survived
100 C 25 min1. Paecilomyces variotii and some strains of Fusarium spp. May survive to thermal
treatments at 95 C per 10–20 s1, probably due to the existence of structures as clamidosphores.
Some strains of P.variotii had been isolated from deteriorated products pasteurized at 93 C for 5
min. Although in low numbers (<10 cfu ml1 in the final product after storage for >90 days at 28
C), P. variotii was isolated from apple nectar, showing slow germination. The heat resistance and
the effects of continuous pasteurization on the inactivation of B. fulva ascospores in clarified
apple juice were also studied. Three different strains: Byssochlamys fulva IOC 4518, B. nivea ATCC
24008, and B. nivea FRR 4421 were tested. Byssochlamys fulva IOC 4518 was the most heat-
resistant strain and was eliminated only by a heat treatment of 100 C 5 min1.

Heat Tolerance of HRM Isolated from Juices

Heat resistance of N. fischeri ascospores is affected by sporulation and the type of heating medium
used. Ascospores of three strains of N. fischeri were grown on three different sporulation media
(Fowell’s acetate agar, apple juice agar, and grape juice agar) were evaluated for heat resistance
in apple juice, grape juice, and 0.1 M potassium phosphate buffer (pH 7.0). The type of sporulation
medium did not affect the heat resistance of ascospores. Ascospores of all three strains at an
initial viable population of w106 cfu ml1 survived at 84 C 120 min1. The rate of thermal
inactivation of ascospores was lower in apple juice than in grape juice or phosphate buffer. Rates
of inactivation (82 C) increased as the pH (2.5, 3.0, and 3.5) of heating media containing fumaric,
citric, tartaric, and acidic acids decreased. The influence of organic acids on heat-resistance
characteristics of Talaromyces flavus ascospores during and after exposureto elevated
temperatures was studied. Fumaric, sorbic, and benzoic acids were clearly more lethal than
acetic, malic, citric, and tartaric acids, and lethality was enhanced as the pH of the heating medium
was reduced from 5.0 to 2.5. Studies about the combined effects of pressure–temperature on heat
resistance of ascospores of B. nivea, B. fulva, N. fischeri, and T. flavus at 20, 50, and 60 C were
conducted. At 20 C, a 9000 bar treatment for 20 min completely inactivated all T. flavus
ascospores (decimal reduction (DR) 3.5 from the initial population) and reduced N. fischeri
ascospores by w2 log cycles, while the ascospores from B. nivea and B. fulva underwent no
10 | P a g e

reduction. In apricot nectar that was preheated at 50 C, all four species were inactivated in 1–4
min by a treatment (3.5–5 DR at 8000 bar), while in an apricot nectar preheated at 60 C, the same
result was obtained in 1–2 min using a 7000 bar pressure. Pressure-resistance of ascospores at
50 and 60 C was found to be lower in distilled water than in apricot nectar. The growth of P.
variotii in pineapple juice was modeled as a function of pH (2.7–3.5), water activity (0.84–0.98),
and natamycin concentration (0–10 ppm). In the absence of natamycin, high values of aw such as
0.98 favored quick germination and growth of the mold in 4 days, with visible colonies,
overcoming the inhibitory effect of pH. For aw 0.91, pH 3.1, and 5 ppm of natamycin, growth as
visible colonies (2 mm diameter) was observed after 39 days of lag phase. The lag phase increased
from 2 days to 40 days when conditions changed from pH (2.7–3.5) and aw 0.98 to pH 3.1, aw
0.91 and 5 ppm natamycin. Thus, hurdle technology may be used to inhibit the germination and
posterior growth of the mold, since at aw 0.84 and natamycin concentration of 10 ppm inhibit the
mold for 90 days. Studies indicate that small variations in temperature (w2 C) during the
concentrated apple juice pasteurization could result in the elimination or survival of HRMs due
to its nonlogarithmic inactivation behavior. Temperature variations could culminate in the
survivor of HRMs in pasteurized juices even when low counts (<10 spores per 100 ml) were
present in the raw materials.
Spore-Forming Bacteria

Alicyclobacillus spp.

Alicyclobacillus spp. is currently one of the microorganisms of concern in the fruit juice industry
(see Alicyclobacillus). Alicyclobacillus includes thermophilic–acidophilic–heterotrophic bacteria
and the term thermo-acidophilic refers to this particular group. The thermophilic and acidophilic
characteristics of Alicyclobacillus spp. allow resistance to current pasteurization processes, and
the ability to produce off-flavors in the product poses potential economic losses for the juice
industry. Most studies concerning Alicyclobacillus spp. related to spoilage are focused on A.
acidoterrestris. However, recent studies have revealed other Alicyclobacillus species as equally
able to cause off-flavors. Recently, other Alicyclobacillus species have been identified:
Alicyclobacillus hesperidum from volcanic soils; Alicyclobacillus herbarius from herbal tea made
from dried flowers of hisbicus; Alicyclobacillus acidiphilus from off-flavor orange juice in Japan;
and Alicyclobacillus pomorum from mixed fruit juice. Alicyclobacillus herbarius is more closely
related to Alicyclobacillus cycloheptanicus in having predominately u-cycloheptane fatty acids in
the cell membranes. Alicyclobacillus acidiphilus is able to produce guaiacol and cause spoilage in
acidic beverages. Alicyclobacillus spp. are soilborne bacteria, and do not strictly require
thermophilic and acidic environments. Alicyclobacillus spp. are Gram-positive, rod-shaped,
thermophilic, and acidophilic spore-forming bacteria. Depending on the different species, growth
temperatures range from 20 to 70 C, with optimum temperatures from 42 to 60 C. Alicyclobacillus
spp. can also grow over a wide pH range, generally reported between pH 2.5 and 6.0. Spore
formation in Alicyclobacillus spp. is terminal or subterminal, with or without swollen
sporangium. The most distinctive characteristic of Alicyclobacillus spp. is the presence of u-
alicyclic fatty acids as the major membrane component. Researchers suggest that u-alicyclic fatty
acids are associated with the exceptional resistance of Alicyclobacillus spp. to acidic conditions
and high temperatures. It was demonstrated that u-cyclohexane fatty acid–containing lipids pack
densely, resulting in low diffusion at high temperatures. Closely packed rings of the u-alicyclic
fatty acids may form a protective coating for the cell membrane, and contribute to the resistance
of this species to acidic conditions and high temperatures. The major off-flavors associated with
the spoilage caused by Alicyclobacillus spp. can be divided into two groups: guaiacol and the
halophenols, including 2,6-dibromophenol (2,6-DBP) and 2,6-dichlorophenol (2,6-DCP).
Although guaiacol generally is accepted as the predominant metabolite associated with the smoky
11 | P a g e

taints in fruit juices, the importance of 2,6-DBP and 2,6-DCP should not be overlooked. The factors
that affect guaiacol production are Alicyclobacillus concentration, inoculation temperature, and
heat shock. Guaiacol was detected in orange juice and apple juice when 105 cfu ml1 of A.
acidoterrestris were present. In relation to temperature, it is hypothesized that the reaction rate
of guaiacol production increases as incubation temperature increases. For Alicyclobacillus spp. to
produce guaiacol, vegetative cells must be present rather than dormant spores. Activation is the
process of conditioning spores to germinate. Among the various activation methods available,
exposure to sublethal heat is used most commonly at lab and industrial conditions. Several heat-
shock recommendations have been reported for Alicyclobacillus spp. In studies, a 100%
population increase was observed when spores were heat shocked at 60 C for 30 min. The
greatest recovery was reported at 70 C for 20 min. On a worldwide basis, the incidence of
Alicyclobacillus spp. has been reported mainly in apple and orange juices or in acidic beverages,
but also from mango juice and pear concentrate. The presence of thermo-acidophilic spores was
quantified in 57 samples of commercial passion fruit juice. Sixteen (28%) samples were positive,
and the number of spores in positive samples ranged from 1.1 to 23 NMP 100 ml1 of juice. Higher
incidence occurred during June and July (dry seasons). Various commercial drinks have been
evaluated for their ability to support the growth of Alicyclobacillus spp. Apple–orange–pineapple
(pH 2.9, 14.8 Brix), grapefruit (pH 3.2, 10.4 Brix), orange (pH 3.6, 12.0 Brix), and pineapple (pH
3.3, 13.4 Brix) supported one of the tested isolates. Cranberry (pH 2.4, 14 Brix), apple–grape (pH
2.8–3.7, 12.2–14.8 Brix), apple–grape–cherry (pH 3.7, 12.4 Brix), and prune juice (pH 3.7, 18.8
Brix) did not support growth, nor did Concord grape juice at pH 2.9 or 3.3. The soluble solids
concentration of juices is an important growth factor for Alicyclobacillus spp. Growth was
inhibited when sugar content in the juice samples exceeded 18 Brix. Phenolic compounds may
also influence the growth of the species since red juice was found to be more inhibitory than white
juice. Ethanol prevented growth when concentrations exceeded 6%. Spoilage of pasteurized fruit
juices by Alicyclobacillus spp. presents a considerable challenge to the food industry. Parameters
such as storage temperature may inhibit the growth of Alicyclobacillus spp. or stimulate the
germination of its spores. Temperature studies indicate that storage of commercial pasteurized
fruit juices at temperatures below 20 C is likely to prevent germination and outgrowth of spores
and may provide a potential control measure for the industry to avoid spoilage by Alicyclobacillus
spp.

Thermal Resistance of Alicyclobacillus spp.

Alicyclobacillus spores resistance is not affected significantly by pH reduction of the heating
medium. z-Values remained the same over the pH range tested (pH 3.0–8.0). It was reported that
under low pH conditions, spores of A. acidoterrestris exhibit stronger Ca2þ and Mn2þ binding
capacity than that of other Bacillus species tested. Little change in Ca-DPA concentration and the
strong ability to bind divalent ions in A. acidoterrestris spores are related to their specific heat
resistance. The D95 C values reported for A. acidoterrestris in orange, apple, mango, and cupuaçu
ranged from 2.7 to 3.6, 2.3 to 2.8, 8.3, and 2.8 min, respectively, and the z-values for these juices
can range from 7.7 C in apple juice to 21.3 C in mango pulp. Nevertheless, in McKnight et al.
(2010), D95 C and z-values of Alicyclobacillus isolated from passion fruit juice was determined.
For three A. acidoterrestris strains (DSM 2498, E14, E25, and E27) (Table 2), values were
marginally inferior (D95 C and z of <2 min and 7.6 C, respectively) to those previously described
in the literature for A. acidoterrestris in fruit juices (pH 3.5 and 13 Brix). These differences are
the result of the main factors influencing the heat resistance of microorganisms, such as the strain
under study, the temperature and time of incubation, and the heating medium characteristics.

Growth Modeling of A. acidoterrestris and the Effect of Processing and Storage Conditions
12 | P a g e

It was examined the effect of five different cooling abuse conditions on A. acidoterrestris growth
in hot-filled (92 C for 10 s) orange juice spiked with either 102 or 103 spores ml1 of juice. Growth
curves were followed for the survivors. It was found that only with storage at 20 C did the
population remained inhibited during the 6 months of orange juice shelf life. Alicyclobacillus
acidoterrestris predicted growth parameters were influenced significantly (p < .05) either by
inoculum level or cooling and storage conditions. The time required to reach a 104 cfu ml1
population of A. acidoterrestris during storage was considered to be an adequate parameter to
indicate orange juice spoilage by A. acidoterrestris due to detectable guaicol production.
Therefore, hot-filled orange juice should be stored at or below 20 C for microbial stability. It was
concluded that A. acidoterrestris spores would survive the hot-fill process since this
pasteurization process causes <1 log reduction and that spores were able to germinate and spoil
orange juice in a few days (5–6 days) when the final storage temperature was 35 C. Predictive
modeling showed that only the maximum population ratio (K) and the time to reach 104 spores
ml1 were affected by spore inoculum size (p <.05). Manipulation of more than one factor (hurdle
technology), as well as the use of antimicrobial, can be an alternative to prevent the development
of A. acidoterrestris in orange juice, thus, contributing to increase its shelf life. The adaptation
time of A. acidoterrestris CRA 7152 in orange juice was studied as a response to pH (3–5.8),
temperature (20–54 C), Brix (11–19) as well as nisin concentration (0–70 IU ml1) effects. Baranyi
and Roberts’ model (1995) best described the experimental data. Inhibition of bacteria was
obtained through several studied combinations for at least 47 days of storage. The shortest period
of adaptation was observed between 37 and 45 C incubation temperature and pH between 4 and
5, yet the longest periods of adaptation could be obtained around 20 C with pH close to 3.0.
Statistical analysis of the quadratic model showed that the adaptation time increased as
temperature or pH decreased, and as nisin concentration or soluble solids increased. Another way
to inactivate A. acidoterrestris spores in apple juice (without chemical compounds) is combining
heat treatment (90 C) and high pressure (414–621 MPa min1). This combination may reach >5.5
DR of this microorganism. In a different approach, enterocin AS 48 (2.5 mg ml1) was used to
inactivate vegetative cells and spores of Alicyclobacillus acidocaldarius and three strains of A.
acidoterrestris.

Other Spore-Forming Bacteria

The involvement of other spore-forming bacteria in the spoilage of fruit juices is less pronounced
than that recently associated with Alicyclobacillus. Besides phenolic or medicinal off-flavors,
other taints mainly described as ‘musty,’ ‘like roots,’ and ‘earth,’ also play an important role in the
spoilage of fruit juices These metabolites are produced by Actinomycetes (Streptomycetes),
which are ubiquitous microorganisms, accounting for a large part of microbial community in soils.
Actinomycetes are rod-shaped, aerobic Gram-positive bacteria that produce spores and form
long, thread-like branched filaments. It seems reasonable that these bacteria may be introduced
into juices by poorly washed fruits that are contaminated with soil. The spores of thermophilic
13 | P a g e

Actinomycetes can be highly resistant to high temperatures, surviving in sucrose solution up to

100 C between 10 min and 4 h. In some instances, Bacillus licheniformis, Bacillus polymyxa,
Bacillus subtilis, and B. coagulans have caused spoilage of acid and acidified foods. Growth of
these species in acid foods, such as tomatoes and tomato juice, is of concern, because some strains
can raise the pH of the product onto the range at which growth of Clostridium botulinum is
possible. On occasion, mesophilic aerobic sporeformers may be recovered from, or cause spoilage
of, acidified food and fruit drinks, since such products normally receive only a hot-fill-and-hold
or pasteurization heat treatment. Bacillus coagulans, C. pasteurianum, Clostridium butyricum,
and Thermoanaerobium thermosaccharolyticum are important in respect to spoilage problems
in tomato juice and other tomato products. The butyric anaerobes are capable of germination and
growth at a pH 4.2–4.4 and are significant in the spoilage of nonpressure-processed acid food,
such as tomatoes and tomato products, and particularly if the pH is above 4.2. They are also of
spoilage significance in underprocessing of other acid and acidified foods and occasionally in low-
acid canned foods as well.

Pathogenic Bacteria
Fruit juice–associated outbreaks of illness reported to the US Centers for Disease Control and
Prevention Foodborne Outbreak Reporting System has been reviewed. From 1995 through 2005,
21 juice-associated outbreaks were reported; 10 implicated apple juice or cider, 8 were linked to
orange juice, and 3 involved other types of fruit juice. These outbreaks caused 1366 illnesses, with
a median number of 21 cases per outbreak. Among the 13 outbreaks of known etiology, five were
caused by Salmonella, five by E. coli O157:H7, two by Cryptosporidium, and one by Shiga toxin-
producing E. coli O111 and Cryptosporidium. Fewer juice-associated outbreaks have been
reported since the hazard analysis and critical control points (HACCPs) regulation for juice was
implemented. In general, the risk of juices is lower if compared with other foods such as meat and
seafood, with the exception of cases of unpasteurized products. Although rare, in 2006, five
people in the United States (Georgia and Florida) and Canada (Toronto) became violently sick
and one victim died after drinking unpasteurized carrot juices. An investigation conducted by the
Food and Drug Administration (FDA) revealed that the juice was contaminated with C. botulinum.
The investigation revealed that due to improper refrigeration the microorganism may have
grown and produced toxin, since the product has a pH >4.6. To prevent future botulism outbreaks
caused by contaminated nonpasteurized higher pH juices, the FDA is considering ordering
acidification of such products produced in the United States. The pathogens most implicated in
outbreaks caused by fruit juices were E. coli strains and Salmonella. Of the diarrheagenic E. coli,
the enterohemorrhagic (EHEC; see Escherichia coli Enterohemorrhagic E. coli (EHEC), Including
zzANon-O157) one (E. coli O157:H7; see Escherichia coli Enterohemorrhagic E. coli (EHEC),
Including Non-O157) is the main concern, due to its low infective doses and its association with
hemorrhagic colitis, thrombotic thrombocytopenic purpura, and the hemolytic-uremic
syndrome, which results in 3–5% mortality among infected children. Salmonella has been
associated with outbreaks caused by juices in the past three decades and the incidence normally
was associated with poor hygiene of the food handlers and with the presence of acid-tolerant
serovars. The major problem associated with acid-adapted Salmonella is its great ability to
survive gastric fluid conditions, resulting in higher incidence of salmonellosis. One of the
requirements of FDA regulation is to attain at least 5-log reductions of the target pathogen
microorganism on the juice pasteurization process so the commercial thermal process has to be
verified and checked to conform to this regulation. Listeria monocytogenes is not well established
as a relevant fruit juice–borne pathogen in international literature, as compared with Salmonella
and E. coli 0157:H7. This pathogen can be considered to be of concern in fresh fruits and fruit
juices, however, due to its ability to survive under a variety of adverse conditions. Listeria
14 | P a g e

monocytogenes is able to survive and grow on equipment surfaces and presents a markedly
psychotropic behavior. Pathogens control in juices have been studied using nonthermal
treatment, which ensure the sensorial properties of juices. Pulse electric field treatments have
been reported to extend the shelf life of orange juice and apple juice, achieving a 5-log reduction
for pathogens, including E. coli O157:H7. In addition, ultraviolet (UV) light treatment has been
reported to be effective in the reduction of pathogens and spoilage microorganisms in juices.
Effectiveness varied with flow rate and exposure. A sound HACCP program in conjunction with
UV light treatment is recommended for juice producers. Irradiation of apple juice with 1.8 kGy is
sufficient to achieve 5DR inactivation of E. coli O157:H7. This treatment has been recommended
by FDA.

Protozoa
Cryptosporidium cayetanensis and Cryptosporidium parvum have been shown to be potential
contaminants and emergent pathogens associated with juices and ciders being the cause of
cristosporidiosis. Cryptosporidium parvum were the etiological agent in outbreaks in the United
States due to consumption of unpasteurized apple juice and apple cider. In the Brazilian Amazon
region, the appearance of the Chagas disease caused by Trypanossoma cruzi has been associated
with the ingestion of triatomine vectors or their excrements that can be crushed together with
the fruit during açai palm juice preparation. Similar cases were reported to have occurred with
bacaba juice and sugarcane juice. Between 1968 and 2005, an average of 12 cases per year of
Chagas diseases was reported in the north of Brazil.

Virus
Enteric viruses may be present on fruits, as a result of human fecal contamination either before
or after harvest. Noroviruses (formerly Norwalk and Norwalk-like viruses), small round-
structured viruses, and hepatitis A virus are the major concerns. Incorrect handling of fruits
during preparation for consumption is an important cause of contamination, but workers and the
use of polluted water in the production line may also be sources of contamination. There have
been few studies of the survival of hepatitis A or noroviruses in juices; however, the major
influences on survival of pathogenic bacteria also determine the survival time of enteric viruses.
Some substances present in fruits cause reversible inactivation of viruses. Some researchers cited
that hepatitis A has been associated with the consumption of lettuce, raspberries, frozen
strawberries, berries, and orange juice. Virus control is hard because the organisms remain
infectious after refrigeration and freezing, which can also facilitate the persistency in the product.
Chlorination of water to wash fruits and vegetables has been recommended to inactivate Norwalk
and hepatitis A viruses present in the surfaces using 10 and 5 mg l1, respectively. Other authors,
however, reported that these viruses are resistant to the majority of disinfectants. In addition,
thermal treatment in boiling water is the most effective way to inactivate viruses, while the
efficacy of mild heat treatments is not guaranteed. Therefore, the best way to control juice
contamination from viruses is by applying the principles of good manufacturing practices
(including hygienic) and HACCP, especially at the primary production level.
15 | P a g e

Contamination of Vegetable Beverages by Microorganisms

Few studies were found related to the contamination of vegetable beverages by spoilage or
pathogen microorganisms. Bacillus coagulans, C. pasteurianum, C. butyricum, and T.
thermosaccharolyticum are important in respect to spoilage problems in tomato juice and
products or in other acidic products. Tomato juice is more prone to spoilage by these
microorganisms due to its typical pH of 4.3, which is favorable for the growth of these spoilage
microorganisms. The thermal characteristics of the spores and vegetative cells of B. coagulans
ATCC 8038 in tomato juice have been evaluated as a function of temperature (95–115 C). The z-
values varied from 8.3 to 8.7 C. Sporolactobacillus are Gram-positive rods, microaerophilic,
negative catalase, mesophilic spore-formering bacteria that present moderate heat resistance.
This microorganism may be responsible for the undesirable fermentation of foodstuffs before
further processing. The possibility exists that Gram-positive spore-forming rods isolated from
heat-processed foods may well have been misidentified as Bacillus and Clostirdium species. This
statement applies not only to Sporolactobacillus but also to other spore former microorganisms
that cause flat sour spoilage. Survival and growth patterns of E. coli O157:H7, Salmonella
typhimurium and L. monocytogenes in black carrot juice were studied. The effect of several
parameters, such as concentration, pH, incubation temperature, and incubation time, were
investigated. Listeria monocytogenes has been found to be the less resistant microorganism to
the variable conditions; there were only w1- and 2-log reductions in the number of cells in the
juice samples incubated at 4 C for 2 and 7 days, respectively. Incubating at low temperature (4 C
for 7 days) enhanced the survival of test microorganisms.
16 | P a g e

RESEARCH METHODS
The present study was conducted in laboratory of Biophysics Dept., Institute of Medical Sciences,
Department of Mycology and Plant Pathology, Institute of Agricultural Sciences, and in
Department of Home Science, Mahila Maha Vidyalaya, Banaras Hindu University, Varanasi, Uttar
Pradesh.

SELECTION OF SAMPLES

Samples were taken whose fruit juices are consumed by more population daily. These fruit juices
are consumed by peoples for their health benefit or as prescribed by doctor for therapeutic use.
In the present study, the samples were selected : orange, mosambi, lemon, papaya, mango, green
grapes, purple grapes, apple, pineapple and pomegranate.
PREPARATION OF SAMPLE

The following procedure was used for preparation of sample. Collect fresh fruits, wash it. Peel and
cut it. Squeeze the juice by pressing it by hand with the help of sterilized muslin cloth store the
fruit juices in sterilized beaker. Use every time clean muslin cloth and sterilized beakers for
squeezing and storage of fruit juices.

STORAGE OF SAMPLE
For the storage of sample of fruit juices, 10 sterilized 250 ml beakers were used and these samples
were stored in refrigerator for 48 hrs till the whole study was not carried out.

MEASURING OF PH OF FRUIT JUICES

The pH of fruit juices was measured on fresh (0 hrs), 24 hrs, 36 hrs and 48 hrs duration. pH of
fruit juice was measured by pH meter (Potentiometer).

TITRATION OF FRUIT JUICES

Titration of fruit juices was done by simple titration method (AOAC, 2000).

formula for calculating titrable acidity:

Titration value×Normality of NAOH×Vol.made up×Equivalent weight of citric acid×100

Total acidity (%age) =
Volume taken for titration×Weight of sample×1000

DATA ANALYSIS

For statistical analysis ANOVA was used.

17 | P a g e

RESEARCH FINDINGS AND DISCUSSION

Table 3 shows the effect of keeping time period on pH of fruit juices. Mango juice has maximum
pH (5.3) decreased pH (5.3) which decreased with increase keeping time period with increased
keeping time period(24 hrs) whereas,pomegranate juice had minimum pH (2.6) and there was
no keeping time period effect on pH. Mango juice had maximum(36 hrs). pomegranate juice had
minimum pH (2.6) also decreased with increased keeping time period (36 hrs).Pomegranate juice
had minimum pH(2.8) and showed no effect on pH due to keeping time . Similar findings were
reported by Zahid et al.,(2008) in which pH decreased during storage and the mean values ranged
between 3.89 and 2.92. Minimum decrease in the pH of apple juice was observed during
storage.Data pertaining to acidity (%) in different juices at 0 hours have been given in Table 2.
Screening of data revealed that the percentage of acidity ranged from 0.48 to 9.56. The lowest
total acidity (0.48 %) was found in treatment T8 (apple juice) at 0 hour. Maximum per cent of
total acidity was recorded with treatment T5 (9.56%), which showed its significant over all other
treatments .Treatments T9 gave significantly more acidity in comparison to treatment T2, T3 and
T10. Treatment T6 gave insignificant higher acidity incomparison to T8. Treatment T3 also gave
insignificantly greater acidity per cent than treatment T1. Treatment T2 gave in significantly
more acidity per cent than treatment T3, T4, and T10. Data pertaining to acidity (%) in different
juices at 24 hours has been given in Table 4. Screening of data revealed that the percentage of
acidity ranged from 0.67 to 12.51. The lowest total acidity (0.67 %) was found in treatment
T8(apple juice) at 24 hour. Maximum per cent of total acidity was recorded with treatment T5
(12.51%), which showed its significant superiority over all other treatments.Treatments T9 gave
significantly more acidity in comparison to treatment T2, T3 and T10. Treatment T6 gave
insignificant higher acidity in comparison to T8. Treatment T3 also gave insignificantly greater
acidity per cent than treatment T1. Treatment T2 gave in significantly more acidity per cent than
treatment T3, T4, and T10. Data pertaining to acidity (%) in different juices at 36 hours has been
given in Table 2. The data revealed that the percentage of acidity ranged from 0.98 to 13.36. The
lowest total acidity (0.98 %) was found in treatment T8(apple juice) at 36 hour. Maximum per
cent of total acidity was recorded with treatment T5 (13.36%), which showed its significant
superiority over all other treatments .Treatments T9 gave significantly more acidity in
comparison to treatment T2, T3 and T10. Treatment T6 gave insignificant higher acidity in
comparison to T8. Treatment T3 also gave insignificantly greater acidity per cent than treatment
T1. Treatment T2 gave in significantly more acidity per cent than treatment T3, T4 and T10. Data

pertaining to acidity (%) in different juices at 48 hours have been given in Table 2.The data
revealed that the percentage of acidity ranged from 1.12 to 16.05. The lowest total acidity (1.12
%) was found in treatment T8(apple juice) at 48 hour. Maximum per cent of total acidity was
Table 3: Effect of keeping time period on acidity of fruit juices
No. of sample Sample of fruit pH reading at
juices 0 hr 24 hr 36 hr 48 hr
1. Orange 4.5 4.5 4.75 4.7
2. Mosambi 3.95 4.15 4.2 4.25
3. Green grapes 4.3 4.35 4.45 4.50
4. Purple grapes 4.05 4.2 4.20 4.2
5. Lemon 4.25 4.25 4.35 4.3
6. Papaya 3.5 3.6 3.63 3.67
7. Mango 5.3 5.2 5.25 5.37
8. Apple 3.85 3.85 3.9 3.95
9. Pineapple 3.75 3.75 3.8 3.9
10. Pomegranate 2.6 2.6 2.8 2.8
recorded with treatment T5 (16.05%), which showed its significant superiority over all other
18 | P a g e

treatments .Treatments T9 gave significantly more acidity in comparison to treatment T2, T3 and
T10. Treatment T6 gave insignificant higher acidity in comparison to T8. Treatment T3 also gave
insignificantly greater acidity per cent than treatment T1. Treatment T2 gave in significantly
more acidity per cent than treatment T3, T4 and T10. In T1(orange juice) the acidity began to
increase in 0- 48 hours. Similar findings were reported by Wissanee (2007) where at 40C the
orange juice titratable acidity began to increase in 2-6 days. The increase in TA (titratable acidity)
indicated fermentation of orange juices. Mean value for titratable acidity of T8 (apple juice)
increased from (0.19 to 0.64 %) during keeping time period. Similar findings were reported by
Zahid et al.(2008). Mean values for titratable acidity of apple juice increased from 0.34 to 0.53
per cent during storage. T3 (green grapes),T4 (purple grapes), T5 (lemon), T6 (papaya), T9
(pineapple) and T10 (pomegranate) juices, their mean for titratable acidity regularly increased
during the keeping time period, whereas, in T2 (mosambi) and T7 (mango) juice, their titratable
acidity decreased after 24 hrs but after that there was noted regularly increased of titratable
acidity during the keeping time period.The number of micro-organisms in fruit juices can affect
the microbial quality of fresh fruit juices. A specific research related to this issue was done to
quantify the transfer rates of microorganisms during an extraction process. Aspergillus niger,
Aspergillus flavus and species of Mucor and Rhizopus were detected from fruit juices of different
fruits (Table 3). It was found that about 1.7-2.6 per cent of total aerobic organisms and 2.3- 2.6
per cent of acid uric organisms from the washed fruits were introduced into the fruit juices during
extraction process. The quality of fresh fruit juices is essentially depending on careful fruit
handling and strict processing sanitation (Steven and Davis,2001).The result in the present study
clearly indicated the poor hygienic conditions of these fruit juices and the consumers are at risk
of contacting food borne infections.

Table 4: Effect of keeping time period on titratable acidity of fruit juices sample
Sample(fruit juises) Treatment 0 hours 24 hours 36 hours 48 hours
Orange T1 0.94 0.81 1.11 1.88
Musambi T2 1.41 0.94 2.09 2.52
Green grapes T3 1.06 1.60 2.05 2.15
Purple grapes T4 1.40 1.60 2.50 2.90
Lemon T5 9.56 12.51 13.36 16.05
Papaya T6 0.56 0.73 1.07 1.24
Mango T7 1.92 1.32 2.35 3.12
Apple T8 0.48 0.67 0.98 1.12
Pineapple T9 2.08 2.37 2.50 2.59
Pomegranate T10 1.20 1.41 1.96 2.22
S.E.± 0.19 0.10 0.07 0.06
C.D. (5%) 0.39 0.21 0.15 0.13

Table 5: Identification of fungus in different fruit juices

Sample (fruit juices) Name of fungi
Orange, Pomegranate Aspergillus niger, Aspergillus flavus, Mucor sp.
Mosambi, Green grapes, Purple Grapes, Aspergillus niger, Aspergillus flavus
Lemon, Mango, Apple, Pineapple
Papaya Aspergillus niger, Aspergillus flavus, Rhizopus sp.
19 | P a g e

CONCLUSION
It has been concluded that pH of fruit juices decreased during storage. Keeping time period affects
the titratable acidity of different fruit juices. Titratable acidity increased with increasing keeping
time period. It is contended that contamination is mainly due to poor quality of water used for
dilution as well as prevailing unhygienic conditions related to washing of utensils, contaminated
water and ice, poor personal and domestic hygiene, peeling of fruits before hand washing, dust
particles in the premises. The practice of consuming fresh fruit and juices cannot be stopped on
unhygienic grounds people should avoid street fruit juices. Regular monitoring of the quality of
fruit juices for human consumption must be introduced to avoid any future pathogenic outbreak.
20 | P a g e

REFERENCES
Amato, D. (1999). The mineral content of bottled water and other beverages: Implications for
health and disease. American J. Med., 105: 125-130.
AOAC (2000). Official method of analysis. Association of Official Analytical Chemist, Washington,
D.C.,U.S.A.

Dietary Guidelines for Americans (2005). United States Department of Agriculture Science. US,
pp. 425–430.

Fasoyiro, S.B., Ashaye, O.A., Adeola, A. and Samuel, F.O. (2005). Chemical and storability of
fruits-flavored (Hibiscus sabdariffa) drinks. World J. Agric. Sci., 1: 165-168.
Fawole, M.O. and Osho, B.A.(2002). Laboratory manual of microbiology. Spectrum Books Ltd.,
Ibadan, 6-45pp..

Ryu, J.H. and Beuchat, L.R. (1998). Influence of acid tolerance responses on survival, growth and
thermal cross-protection of Escherichia coli O157:H7 in acidified media and fruit juices.Internat.J.
Food Microbiol., 45:185-193.
Sandeep, M., Dwarker, A. and Abhijit, G. (2001). Microbiological analysis of street vended fresh
sqeezed corrot juice in Patiala city. Indian. J. Food Safety, 15: 1-3.

Splittstosser, D.F. (1979). Fruits and fruit products. In: Food and beverage mycology, Beuchat,
L.R. (Ed.). Avi Publishing Co., Wedtport, USA., pp. 215-220.
Steven, P. and Davis, C.L. (2001). Transfer of natural and artificially inoculated microorganisms
from orange fruit to fresh juice during extraction. Lebensmittel-Wissenschaft and Technologie, 34:
113–7.
Uljas, H.E. and Ingram, S.C. (1998). Survival of Escherichia coli O157:H7 in synthetic gastric fluid
after cold and acid habituation in apple juice or trypticase soy broth acidified with hydrochloric
acid or organic acids. J. Food Prot., 61: 939-947.
Wissanee and Pinthong Renu (2007). Physical, chemical and microbiological changes during
storage of orange juices cv. Sai Nam Pung and cv. Khieo Waan in Northern Thailand. Internat. J.
Agric. & Biol., 9(5): 726-730.
Zahid Mehmood, Alam Zeb, Mohammad Ayub, Nizakat Bibi, Amal Badshah and Ihsanullah
(2008). Effect of pasteurization and chemical preservatives on the quality and shelf-stability of
apple juice. American J. Food Technol.,3(2):147-153.
Castella, M.L.A., Matasci, F., Schmidt-Lorenz, W., 1990. Influence of age, growth medium, and
temperature on heat resistance of Byssochlamys nivea ascospores. LWT – Food Science and
Technology 23, 404–411.
Dijksterhuis, J., Samson, R.A., 2006. Activation of ascospores by novel food preservation
techniques. In: Hocking, A.D., Pitt, J.I., Samson, R.A., Thrane, U. (Eds.), Advances in Food Mycology.
Advances in Experimental Medicine and Biology, vol. 571. Springer, New York, pp. 247–260.
Ferreira, E.H.R., Masson, L.M.P., Rosenthal, A., Souza, M.L., Tashima, L., Massaguer, P.R., 2011.
Termorresistência de fungos filamentosos isolados de néctares de frutas envasados
assepticamente. Brazilian Journal of Food Technology 14, 164–171.
21 | P a g e

Gumerato, H.F., 1995. Desenvolvimento de um programa de computador para identificação

dealguns fungos comuns em alimentos e determinação de resistência térmica de Neosartorya
fischeri isolado de maçãs. Master thesis. Universidade Estadual de Campinas UNICAMP,
Campinas, S.P. Brazil.

Massaguer, P.R., 2004. Final Report Project: Segurança Microbiológica de Sucos & Drinks
Envasados Assepticamente, 09/1999 a 12/2004, EMBRAPA - PRODETAB, Tetra Pak Ltd.,
MVEngenharia, ABTN. Processo 03-02/98.

Rajashekara, E., Suresh, E.R., Ethiraj, S., 1996. Note: influence of different heating media on
thermal resistance of Neosartorya fischeri isolated from papaya fruit. Journal of Applied
Bacteriology 81, 337–340.

Salomão, B.C., Slongo, A.P., Aragão, G.M.F., 2005. Heat resistance of Neosartorya fischeri in various
juices. LWT/Food Science and Technology 40, 676–680.

Sant’Ana, A.S., Rosenthal, A., Massaguer, P.R., 2009. Heat resistance and the effects of continuous
pasteurization on the inactivation of Byssochlamys fulva ascospores in clarified apple juice.
Journal of Applied Microbiology 7, 197–209.
Scott, V.N., Bernard, D.T., 1987. Heat resistance of Talaromyces flavus and Neosartorya fischeri
isolated from commercial fruit juices. Journal of Food Protection 50, 18–20.
Slongo, A.P., Aragão, G.M.F., 2007. Avaliação da Resistência térmica de Byssochlamys nı´vea e de
Neosartorya fischeri em suco de abacaxi. Boletim CEPPA 25, 217–224.
22 | P a g e

FURTHER READING
Arias, C.R., Burns, J.K., Friedrich, L.M., Goodrich, R.M., Parish, M.E., 2002. Yeast species associated
with orange juice: evaluation of different identification methods. Applied and Environmental
Microbiology, 1955–1961.

Burnett, S.L., Beuchat, L.R., 2000. Review: human pathogens associated with raw produce and
unpasteurized juices and difficulties in decontamination. Journal of Industrial Microbiology and
Biotechnology 25, 281–287.

Centers for Disease Control and Prevention (CDC), 1996. Outbreak of Escherichia coli O157:H7
infections associated with drinking unpasteurized commercial apple juice – British Columbia
California Colorado and Washington October 1996. MMWR Morbidity Mortality Weekly Report
45, 975.

Chang, S., Dong-Hyun, K., 2004. Alicyclobacillus spp. in the fruit juice industry: history,
characteristics and current isolation/detection procedures. Critical Reviews in Microbiology 30,
55–74.

Degirmenci, H., Karapinar, M., Karabyikli, S., 2012. The survival of E. coli O157:H7, S. typhimurium
and L. monocytogenes in black carrots (Daucus carota) juice. International Journal of Food
Microbiology 153, 212–215.

Goto, K., Yokota, A., Fujii, T., 2007. Alicyclobacillus: Thermophilic Acidophilic Bacilli. Springer,
Japan, p. 160.

Massaguer, P.R., Pacheco, C.P., Atarassi, M., Peña, W.L., Gonçalves, A.C., Paula, N.A., Geraldini, L.E.,
Liossi, L.L., Gagliazzi, M.R., Guerra, V.A., 2002. Sensibility and specificity methods of
Alicyclobacillus detection and quantification. Fruit Processing 12, 478–482.

McKnight, I.C., Eiroa, M.N.U., Sant’Ana, A.S., Massaguer, P.R., 2010. Alicyclobacillus acidoterrestris
in pasteurized exotic Brazilian fruit juices: isolation, genotypic characterization and heat
resistance. Food Microbiology 27, 1016–1022.

Nakagawa, H., Hara-Kudo, Y., Onoue, Y., Konuma, H., Fujita, T., Kumagai, S., 1999. Method for
isolation of Escherichia coli O157:H7 from radish sprouts: a collaborative study. Biocontrol
Science 4, 45–49.

Pacheco, C.P., Massaguer, P.R., 2004. Biological validation of tomato pulp continuous heat process.
Journal of Food Process Engineering 27, 449–463.
Parish, M.E., 1997. Public health and nonpasteurized fruit juices. Critical Reviews in Microbiology
23, 109–119.

Peña, L.W., Faria, A.J., Massaguer, P.R., 2004. Development of a predictive model on the growth of
the spoilage mould, Paecilomyces variotii, in pineapple juice. Fruit Processing 6, 420–426.

Put, H.M.C., De Jong, J., 1982. Heat resistance studies of yeasts; vegetative cells versus ascopores:
erythromycin inhibition of sporulation in Kluyveromyces and Saccharomyces species. Journal of
Applied Bacteriology 53, 73–79.

Renard, A., Marco, P.G., Egea-Cortines, M., Weiss, J., 2008. Application of whole genome
amplification and quantitative PCR for detection and quantification of spoilage yeasts in orange
juice. International Journal of Food Microbiology 126, 195–201.
23 | P a g e

Sharma, M., Adler, B.B., Harrison, M.D., Beuchat, L.R., 2005. Thermal tolerance of acid-adapted and
unadapted Salmonella Escherichia coli O157:H7 and Listeria monocytogenes in cantaloupe juice
and watermelon juice. Letters in Applied Microbiology 41, 448–453.

Silva, A.R., Santa’Ana, A.S., Massaguer, P.R., 2010. Modelling the lag time and growth rate of
Aspergillus section Nigri IOC 4573 in mango nectar as a function of temperature and pH. Journal
of Applied Microbiology, 1105–1116.

Spinelli, A.C.N.F., Sant’Ana, A.S., Pacheco, C.P., Massaguer, P.R., 2010. Influence of the hot-fill
water-spray-cooling process after continuous pasteurization on the number of decimal
reductions and on Alicyclobacillus acidoterrestris CRA 7152 growth in orange juice stored at 35
C. International Journal of Food Microbiology 137, 295–298.

Yuk, H.G., Schneider, K.R., 2006. Adaptation of Salmonella ssp. in juice stored under refrigerated
and room temperature enhances acid resistance to simulated gastric acid. Food Microbiology 23,
694–7
24 | P a g e