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Pathogenic micro-organisms in poultry and eggs Prevention of contamination of poultry meat, eggs and egg products Edited by A. Franchini, R.W.A.W. Mulder ACr 0 o^ '°*,> ' S - i t a . to*·" Rome, Italy, 23 to 25 October 1996 EUROPEAN COMMISSION EUROPEAN COMMISSION COST Action 97 Patho...

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Significance of ante mortem indicators on turkey meat inspection Carlo D'Ascenzi

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Pathogenic micro-organisms in poultry and eggs

Prevention of contamination of poultry meat, eggs and egg products Edited by A. Franchini, R.W.A.W. Mulder

0 o^

ACr

'°*,>

' S - i t a . to*·"

Rome, Italy, 23 to 25 October 1996 EUROPEAN COMMISSION

EUROPEAN COMMISSION COST Action 97 Pathogenic micro-organisms in poultry and eggs

Prevention of contamination of poultry meat, eggs and egg products Proceedings of a workshop held ¡n Rome, Italy, 23 to 25 October 1996 Organised by Istituto Superiore di Sanità

Edited by: A. Franchini Istituto di Zootecnica Facoltà di Agraria University of Bologna Italy R. W. A. W. Mulder DLO Institute for Animal Science and Health Lelystad Netherlands

A great deal of additional information on the European Union is available on the Internet. It can be accessed through the Europa server (http://europa.eu.int). Cataloguing data can be found at the end of this publication. Luxembourg: Office for Official Publications of the European Communities, 1998 ISBN 92-828-1997-3 © European Communities, 1998 Reproduction is authorised provided the source is acknowledged. Printed in Italy PRINTED ON WHITE CHLORINE-FREE PAPER

CONTENTS INTRODUCTION

i

W. F. Jacobs-Reitsma, N. M. Bolder and R.W. A. W. Mulder (NL) The influence of pre-slaughter stresses on the incidence and extent of human pathogens in poultry

3

G. Pezzotti, G. Migliorati, T. Toscani, P. Semprini and A. M. Conte (I) Influence of transportation stress on Salmonella spp. contamination in broiler carcasses

7

C D'Ascenzi, D. Gianfaldoni and P. Gaspari (I) Significance of ante-mortem indicators on turkey meat inspection

11

P. Colombo (I) Community provision in poultry processing: actual situation in Member States (European Commission)

15

L. Ellerbroek, Th. Janssen, P. Krause and E. Weise (D) Process control in poultry cutting plants

17

S. Ferrarini, F. Mutinelli and A. Ricci (I) Meat inspection methodology in a poultry processing plant

23

A. P. Salinetti, G. Fontanella, S. Bilei, A. I. Bozzano, M. Mari and S. Saccares (I) Isolations of salmonella strains and hygienic and sanitary controls of poultry meat on the market from 1990-95

29

K. Vareltzis, N. Soultos, P. Koidis, J. Ambrosiadis and C. Genigeorgis (EL) Antimicrobial effects of sodiumtripolyphosphate against bacteria attached to the surface of chicken carcasses

33

M. Sudbeck (D) Influence of scalding time on microbiological quality of chicken

39

F. Mates and K. Kovarik (CZ) Comparison of salmonella findings in slaughtered poultry under the systems of cooling with water or air

43

A. Unanue, S. Ferrarini, N. Ferro-Milone, P. Caccin, A. Ponzoni and A. Ricci (I) Salmonella contamination rates at different sites of a poultry processing plant

47

G. A. Alfredsson, J. Y. Xiang, H. M. Gudmundsson and F. Georgsson (IS) Screening of foods for salmonella and E. coli 0157 by IMS

53

H. Hupkes (NL) Slaughtering equipment and poultry meat hygiene

55

T. J. Humprey (UK) Hygiene in egg production

59

N. M. Bolder (NL) Hygienic practices in egg industry

63

A. M. Prete, L. Vellucci and S. Squarcione (I) Foodborne diseases' outbreaks: data from the national reporting system of infectious diseases

67

F. Sisak and M. Faldynova (CZ) Effect of peracetic acid on salmonella devitalisation in egg shells

73

P. Braun, K. Fehlhaber, U. Mayer and A. Wicke (D) Investigations on behaviour of Salmonella enteritidis in hens' eggs

77

D. De Medici, S. Di Pasquale, G. Ciccaglioni, G. Falzi and L. Orefici (I) Comparison between two rapid methods and the conventional cultural methods for the detection of salmonella in eggs

81

P. Colin (F) Influence of processing technologies and storage conditions on contamination of egg products

85

M. E. Guerzoni, R. Lanciotti, A. De Cesare and C. Chaves Lopez (I) High pressure homogenisation of egg products and its effects on the inactivation of salmonella serotypes

87

C. Solano, O. Bourgi, B. Sesma, M. Alvarez, R. Diaz, I. Dorronsorro and C. Gamazo (E) Release of an extracellular matrix from virulent strains of Salmonella enteritidis

95

List of participants

101

INTRODUCTION COST Action 97 'Pathogenic micro-organisms in poultry and eggs' held its second management committee meeting and workshop of 1996 in Rome, Italy. The working group responsible for the organisation of the workshop was working group 2: 'Prevention of contamination of poultry meat, eggs and egg products'. After its first workshop in 1995 (organised in cooperation with the XHth European Symposium on the Quality of Poultry Meat and the Vlth European Symposium on the Quality of Eggs and Eggs Products in Saragossa, Spain), the Istituto Superiore di Sanità, Rome, Italy, hosted this second workshop. Participants from 18 European countries discussed their research during the two-day meeting. This third volume in the series of publications from COST Action 97 contains the papers presented and it is expected that this publication will further contribute to knowledge transfer throughout Europe. We wish to use this opportunity to express our thanks and appreciation for the support of this workshop to the European Commission, Directorate-General XII/BS.l (Dr Ζ. Somogy), the Italian Poultry F ederation, UNA (Mr R. Celadon) and the local organisers at the Istituto Superiore di Sanità (Dr A. Macrì) and at the University of Bologna (Prof. Dr A. Franchini). A. Franchini R. W. A. W. Mulder

THE INFLUENCE OF PRE-SLAUGHTER STRESSES ON THE INCIDENCE AND EXTENT OF HUMAN PATHOGENS IN POULTRY

W.F. JACOBS-REITSMA, N.M. BOLDER and R.W.A.W. MULDER DLO-Institutc for Animal Science and Health P.O. Box 65, 82(H) AB Lelystad, The Netherlands

Abstract Stress is the word often used in complex situations when an explanation is needed for the failure of preventive measures to control spreading of potentially pathogenic microorganisms. In this respect Salmonella and Campylobacter are the organisms of concern. In the literature influence of stress factors are described to occur during fattening, catching and loading, transport and conditioning. Most research work relates to spreading of Salmonella bacteria. This paper reports on the literature in relation to stress factors and spreading of pathogenic microorganisms and on experiments in which the influence of feed withdrawal and transport stress on the Campylobacter contamination of broilers was estimated. Additionally the effect of the application of a single or a multistage scalding process on the Campylobacter contamination of carcasses directly after scalding was studied. No statistical significant differences between the several experimental groups and treatments were estimated. Introduction Poultry meal forms an important component in the diet of the worlds population. Over the years there have been increasing requirements from consumers for foods that are safe, do not need the addition of preservatives and need little or no preparation or cooking. Thus there is a real pressure to produce poultry and poultry meat products that contain minimal numbers of both spoilage and human pathogenic organisms. Human food-borne diseases are considered to be one of the major problems in the modem world and they are an important cause of economic losses due to hospitalization and absenteeism. Salmonella and Campylobacter infections account for the majority of acute cases of human gastro-enterilis. The results of sentinel and population studies, carried out in the Netherlands, demonstrate that Campylobacter bacteria are the most prominent bacterial cause of acute gaslro-enteritis in humans. This seems to be in contrast to traditional reports, where Salmonella always is the top seeded organism. The difference is probably caused by the past (and in some cases continued) use of investigating and reporting systems, that do not include Campylobacter. The reduction of contamination of live birds with Salmonella and other potentially pathogenic microorganisms has been subject of study in many countries over the last 25 years. Until now no single treatment or process was able to eradicate Salmonella and other pathogens could be identified. Even the mechanism of infection with these organisms could not completely be elucidated. The use of competitive exclusion microflora has been somewhat successful. The treatment of day-old broiler chicks with a microbiota which colonizes them and confers resistance to colonization by pathogens, the so-called competitive exclusion treatment, reduces the contamination of flocks by Salmonella and Campylobacter under laboratory and farm conditions. In studies where flocks were also transported to the slaughterhouse and examined again for these microorganisms, higher contamination rales were found. This is not only true for poultry, similar observations and results are reported from pigs. The collection of poultry on the farm, their transport to the processing plant and the holding time and conditions before slaughter seem to induce the spreading of organisms, resulting in higher contamination and a continued carrier state of the animals. Clinically healthy birds carrying Salmonella and other pathogenic microorganisms may change their excretion pattern of the organisms from intermittent to constant shedding if an external factor upsets the equilibrium of the intestinal flora. A disturbance of the intestinal functions will lower the resistance of the live bird and facilitates the spreading of intestinal bacteria.

Stress is the word often used in those situations which are too complex to be understood and it is often used to explain why preventive measures to control spreading of pathogens in flocks of birds do not work. Stress factors described in literature are observed during fattening, catching and loading, transport and conditioning. On the other hand some stressors are believed to influence positively the resistance to infections with pathogens. Unfortunately this is not described in poultry, but in pigs. So-called social stress, which seems to increase resistance against Staphylococcus aureus and Escherichia colt are described when less animals per square metre are held. Stress in poultry is accompanied by a series of symptoms. The increased corticosteroid levels in blood plasma and the occurrence of damage to the intestinal tract, heart and blood vessels are of major importance. Decreased shear strength of the intestinal tract may results in gut breakage during processing, which is responsible for further spreading of microorganisms over carcasses and equipment (Bugili. 1988). Feed and water withdrawal prior to transport influences gut contents and the emptying of the digestive tract of broilers. Normally chickens empty their caeca every 24 hours, but because of the change in environmental conditions the excretion pattern changes. Some researchers conclude that feed withdrawal minimizes faecal contamination of carcasses (Moran and Bugili. 1990; Papa and Dickens, 1989). Moran and Bugili, 1990. demonstrated that stressing chicken broilers under conditions simulating the practice of feed withdrawal and live haul, results in a delayed caecal retention for another 24 hours. The fact that products became more contaminated with microorganisms when the lime between crating and holding before slaughter increased indicates shedding of faecal material, which spreads over live birds. Also increased Salmonella contamination of slaughtered broilers after transport is reported in literature (Bolder and Mulder, 1983; Moran and Bugili, 1990). The similarity with findings in other parts of the intensive animal industry is striking : Salmonella serotypes after transport observed on slaughtered products originate from live birds or animals, which indicates intestinal origin (Mulder, 1995). In the literature most data relate to spreading or shedding of Salmonella bacteria. Although Campylobacter seems to cause more problems with regard to human public health this prominent position is not yet reflected in the literature. Therefore it was chosen to try to demonstrate the influence of stress factors as feed withdrawal and waiting time before transport and slaughter on Campylobacter contamination of the birds and carcasses after scalding. Additionally the effect of multistage scalding on Campylobacter contamination was studied. Materials and methods (Jacobs-Reitsma, 1997) Broilers Two experiments were carried out. In the first experiment 2 flocks of 400 broilers each were infected with Campylobacter coli C136, Penner serotype 046. In the second experiment 2 flocks of 400 broilers each were infected with Campylobacter jejuni C 356, Penner serotype 0 2 and Campylobacter jejuni C 4021, respectively. Treatments The following treatments were given 1. Ad Lib water/feed/slaughtered without delay 2. Ad Lib water/feed withdrawal 4 hours before slaughter 3. Ad Lib watcr/fced/4 hours crating without water/feed 4. Ad Lib water/feed withdrawal 4 hours before crating/4 hours crating without water/feed The broilers were slaughtered at 42 days of age. Samples and microbiology From individual birds (5 birds per treatment per Hock) caecal contents were plated directly on CCD Agar. In the slaughterhouse, immediately after stunning, killing and bleeding, carcasses were rinsed with 500 ml of physiological saline and after 1 minute of handshaking 1 ml of the rinse fluid was used for the enumeration of Campylobacters. In the second experiment the rinse fluids were also enriched in 10 ml CCD broth.

Results und conclusions The results are summarized in Tables 1, 2 and 3. Table 1. Campylobaeler (Log CFU/g, standard deviation) in caecal contents of broilers without or with teed withdrawal, before and after transport Before Iransport After transport Flock

Ad Lib feed

Feed withdrawal

Ad Lib feed

Feed withdrawal

A H C D NE - not estimated

NE NE 7.6 (0.30) 7.5(0.48)

NE NE 7.9(0.78) 8.0 (0.72)

7.5(0.75) 6.6(1.27) 8.1 (0.63) 7.8 (0.65)

7.8(1.04) 7.7(1.09) 7.1 (0.90) S.3 (0.50)

Table 2. Campylobacter (Log CFU/carcass. standard deviation) in carcass rinse samples of broilers without and with feed withdrawal before and after transport Alter transport Before transport Flock

Ad Lib feed

Feed withdrawal

Ad Lib feed

Feed withdrawal

A Β C D NE - not estimated

NE NE 6.3(1.01) 6.7(1.19)

NE NE 6.8(0.62) 6.9 10.40)

4.9 (0.74) 5.7(1.28) 7.2(1.31) 6.7(1.24)

5.2(1.39) 5.3(1.31) 6.5(0.58) S.l(0.80)

Table 3. Campylobacter (Log CFU/carcass. standard deviation) in carcass rinse samples of broilers without and with feed withdrawal after scalding in one or multistage (three tanks) scaldcrs One scald tank Three stace scald lank Flock

Ad Lib feed

Feed withdrawal

Ad Lib feed

Feed withdrawal

A NE ΝΗ lO'/g from 40 ones (3.0%), E. coli >10\5/g from 42 ones (3.2%). and L monocytogenes from 35 ones (6.67r of the 525 samples examined for this organism) (Table 3). As shown in Table 3. some (0.47%) of the irregular fixids were contaminated, at the same time, by two or more of the considered micro-organisms. As far as Salmonella is concerned, contamination value in the whole period was 5%. but in the last two years it went up to 12.4%. Most of Salmonella positive samples, 53.3%, were poultry-based products: others 23.4% were carcasses of poultry' and bred game and others 23.4% poultry in pieces (Table 3). Salmonella isolates belonged to 20 different serovars: S. sainl-paul (18.2% of isolated Salmonella organisms). S. enteritidis (10.6%) and S. typhimurium (9,1%) resulted the most frequently recovered ones (Table 4).

30

In the 40 S. aureus > l()A3/g contaminated samples were not delected any staphylococcal cnterotoxins. Instead. 11 S. aureus isolates were enlcrotoxm type C producers and another one resulted enterotoxin D producer; all these strains were found in 12 different samples. L monocytogenes was found in 6.6% of the samples (Table 3). whereas L. innocua was isolated from 28% of them. L welshimeri, L gravi and Lseeligeri were less frequent recovered: respectively in 2.17c. 0.9% and 0.2% of the examined foods. The 64.9% of L. monocytogenes strains belonged to 0-1 serotype, the 24.3% to 0-4 type, whereas the 10.8% resulted not typed with both O-l and 0-4 antisera. From L. monocytogenes (L.m.) compulation performed on 24 poultry' samples, resulted that 17 (70.8%) contained 5 O

100000000 1000000 10000 100 1

► Max

i

·

:

I Means

É

Min

2 Plant

Figure 1 : TVC after scalding and plucking Figure 1 illustrates the means, maximum and minimum of the aerobic total viable counts for the four plants. The bacterial data we have found show for each plant a large variation of the bacterial load of the different carcasses after

40

scalding and plucking. But it is easy to sec that both plants using the longer scalding time compared to the other plants show higher means, higher maximum and higher minimum in the bacteriological carcass contamination rates. It is also interesting that all TVC rates higher than 10' were only detected in these plants. But 1 have to add that this high results were always only single carcasses of different flocks.

Discussion The four compared poultry processing plants are using nearly the same equipment. But there are also many individual variations, starting with different chemical data of the used potable water using different plucking finger types etc. Wc also tested always birds from different flocks in the plants never the same flock in the different plants. Because of the high number of unknown possibly influencing factors a statistical evaluation seems not to be efficient. But nevertheless it is conspicuous that both plants using the longer scalding temperatures over all 16 tested flocks showed the higher bacterial contamination rates on the aerobic total viable count. Because of the longer period the carcasses are dragged through the scalding water the contact time between skin and micro-organisms and the time these organisms can affix to the carcasses is longer. This could lead to that higher bacterial load. The higher bacterial contamination rate after scalding and plucking will lead to a higher contamination level at the end of the process (Mead el. al.. 1993). At the beginning of the shelf life lhe birds start on a higher level which will - under the same storing conditions - lead to a shorter shelf life (Mulder, 1982). And indeed plant one produces birds with a longer average shelf life than plant two and three. Because plani four sales the birds frozen, shelf life cannot be compared. During scalding the feathers are loosened so the subsequent plucking procedure is able to remove them (van der Wal and Mulder. 1996). Low temperature (around 50°C) scalded birds are more difficult to pluck than high temperature scalded birds....