Comparison of aerial counts at different sites in beef and sheep abattoirs and the relationship between aerial and beef carcass contamination.

The study examined and compared levels of aerial contamination in commercial beef and sheep plants at four sites, i.e. lairage, hide/fleece pulling, evisceration and chilling. Aerial contamination was determined by impaction and sedimentation onto Plate Count Agar to enumerate Total Viable Counts, MacConkey Agar to enumerate coliforms and Violate Red Bile Glucose Agar to enumerate Enterobacteriaceae. AS I cannot see any difference in the text here - I am not sure what the change is?. The levels of aerial contamination were similar at equivalent sites in beef and sheep plants, irrespective of the sampling method or the type of organisms recovered. Mean log counts recovered on each medium in the chillers were generally significantly lower (P < .05) than the corresponding mean log numbers recovered at the other three sites. The relationship between impaction (air) and sedimentation (surface) counts could be described by the surface to air ratio (SAR) which in this study had an R(2) of 0.77. Further studies in an experimental plant compared counts recovered from the neck of beef carcasses with aerial counts determined by impaction and sedimentation onto agar and irradiated meat pieces. A relationship between counts on beef carcasses and in the air could not be established, irrespective of the method used to compare counts.

Diversity of culturable psychrophilic and psychrotrophic anaerobic bacteria isolated from beef abattoirs and their environments.

This study identified 431 psychrophilic or psychrotrophic isolates from commercial Irish beef abattoir environments and "blown packs" of vacuum-packed beef, using PCR and 16S rRNA sequencing, and estimated their intraspecies genetic diversity using restriction fragment length polymorphism (RFLP) analysis and spacer region PCR (SR-PCR). Twenty-five species were identified in the 431 isolates, with the most frequently recovered species being Clostridium gasigenes (n=315), Clostridium estertheticum (n=17), and a potentially novel species designated strain TC1 (n=52). These species were previously found to be associated with a particular type of spoilage known as blown-pack spoilage (BPS), which occurs in chilled-stored (i.e., -1.5°C to 4°C) vacuum-packaged meat within 2 to 4 weeks and involves the production of large volumes of gas. Overall, the study demonstrates the considerable and not previously reported diversity of the anaerobic microflora in abattoirs and the presence of a wide range of organisms capable of causing BPS at chilled temperatures.

The effect of storage temperature and inoculum level on the time of onset of 'blown pack' spoilage.

AIMS: To examine the effect of storage temperature and inoculum level on the time of onset of 'blown pack' spoilage (BPS) caused by psychrotolerant bacteria in vacuum-packed (VP) meats. METHODS AND RESULTS: Gas-producing species and strains (n = 11), recovered in our laboratory or reported as associated with BPS, were inoculated onto beef or lamb meat pieces at final levels of <10, 10, 10(2) and 10(3) CFU cm(-2), VP and stored at -1.5, 1 or 4 degrees C. Six strains produced observable amounts of gas within 42 days and a further four strains produced gas within 100 days. BPS was observed earliest in VP meats inoculated with Clostridium estertheticum ssp. estertheticum at all inoculum levels/storage temperature combinations examined. Storage temperature and inoculum level significantly affected (P < 0.001 and P < 0.05 respectively) the onset of BPS in all cases. CONCLUSIONS: Controlling contamination levels and lowering the storage temperature delay the onset of BPS. SIGNIFICANCE AND IMPACT OF THE STUDY: The study demonstrates the positive effects of low contamination-low temperature as control interventions preventing/delaying BPS in VP chilled meats and identifies some of the contaminants most likely to cause BPS in chilled stored VP meat products.

The influence of storing beef aerobically or in vacuum packs on the shelf life of mince.

AIMS: To investigate the influence of aerobic or vacuum pack storage of beef trimmings on the microbiology, colour and odour of subsequently produced mince. METHODS AND RESULTS: Trimmings stored aerobically for 7 or 10 days and in vacuum packs for 7, 10, 14 or 22 days at 0 or 5°C were minced, stored aerobically at 0 or 5°C for up to 7 days and examined daily to determine Total viable, Pseudomonas, Lactic acid bacteria, Brochothrix thermosphacta, and Enterobacteriaceae counts, colour and odour. Mincing reduced counts, particularly of Pseudomonas, B. thermosphacta and Enterobacteriaceae, probably because of the action free radicals released from muscle and bacterial cells. Storage of vacuum-packed trimmings for 22 days resulted in improved mince colour and inhibition of the growth of Pseudomonas. CONCLUSIONS: The shelf life of mince from trimmings is directly influenced by the trimmings storage conditions, and longer-term vacuum storage of trimmings produced improvements in mince quality. SIGNIFICANCE AND IMPACT OF THE STUDY: There appears to be no scientific rationale for limiting the storage of vacuum packaging beef trimmings to 15 days, prior to mince production, as stated in EU 835/2004. This study identifies advantages in storing trimmings in vacuum packs for at least 21 days prior to mincing, in terms of improved mince quality.

Detection of numerous verotoxigenic E. coli serotypes, with multiple antibiotic resistance from cattle faeces and soil.

Verotoxigenic E. coli (VTEC) belong to a diverse range of serotypes. Serotypes O157 and O26 are predominately identified in VTEC-associated disease in Europe, however due to difficulty in detection little is known about the epidemiology of non-O157 serotypes. This study reports the identification of 7 VTEC serotypes from cattle faeces and soil. Cattle faeces samples (n=128) were taken from animals in 6 different farms, with soil samples (n=20) obtained from 1 farm. After sample incubation in modified tryptone soy broth (mTSB) supplemented with streptomycin sulphate samples were plated onto sorbitol MacConkey (SMAC) also supplemented with streptomycin sulphate. Bacteria detected on the plates were subjected to biochemical testing, antibiotic resistance profiling, and PCR to detect typical virulence genes, beta-lactamase and class 1 integron associated genes. Serotyping was performed on isolates positive for virulence genes. E. coli was identified from 103 samples, with verotoxin genes present in 7 E. coli isolates. Of these 7 isolates, 5 were resistant to 5 or more antibiotics. The isolate resistant to 9 antimicrobials contained a class 1 integron structure. Serotyping identified 7 separate VTEC, O2:H27, O26:H11, O63:H(-), O148:H8, O149:H1, O174:H21 and ONT:H25. Six of these VTEC have been previously associated with human disease, however with the exception of O26:H11, these serotypes have been rarely reported worldwide. Increased surveillance is required to determine the prevalence of these and other non-O157 VTEC. The presence of multi-antibiotic resistance in these isolates is of concern, and the overall implications for public health must be ascertained.

Effects of reduction in beef surface water activity on the survival of Salmonella Typhimurium DT104 during heating.

AIM: To investigate the influence of reducing beef surface water activity (a(w)) on the survival of Salmonella Typhimurium DT104 during heating. METHODS AND RESULTS: Beef discs were surface inoculated with S. Typhimurium DT104 and either untreated or dried to achieve surface a(w) values of 0.95, 0.85 and 0.70. The samples were vacuum packed, heat-treated at 60 degrees C and removed at predetermined times. The inactivation curves were influenced by a(w) and treatment time. Biphasic inactivation curves were observed for S. Typhimurium DT104 heat-treated on beef samples with altered a(w) values, which were characterized by an initial decline in cell numbers at commencement of heating followed by a much slower rate of inactivation during the remaining treatment period. Point estimates of the heating time required to achieve a 1 log reduction on beef surfaces with a(w) of 0.99, 0.95, 0.85 and 0.70 were 0.5, 1.55, 11.25 and 17.79 min, respectively. CONCLUSIONS: A decrease in beef surface a(w) can substantially enhance the survival of S. Typhimurium DT104 after heating. SIGNIFICANCE AND IMPACT OF THE STUDY: Caution needs to be taken using dry air as a decontamination method as this may rapidly decrease product surface and pathogen a(w) values resulting in enhanced survival.

Isolation and sources of 'blown pack' spoilage clostridia in beef abattoirs.

AIMS: (i) To evaluate methods for isolation and molecular detection of blown pack spoilage (BPS) clostridia and (ii) to survey beef abattoirs for sources and distributions of Clostridium estertheticum and Cl. gasigenes. METHODS AND RESULTS: Molecular detection and conventional isolation methods were used to detect and recover BPS associated clostridia (Cl. estertheticum and Cl. gasigenes), from four commercial Irish beef abattoirs and their environments, during a one year study. DNA-based methods detected 218 Cl. estertheticum and 300 Cl. gasigenes, from 1680 samples, whereas culture-methods only yielded 17 Cl. estertheticum and 176 Cl. gasigenes isolates. BPS Clostridia were frequently detected in beef abattoirs and their environments, especially at areas prior to hide removal. The study noted a higher percentage of positive samples during the month of May (38.6%). CONCLUSIONS: (i) DNA-based techniques are the most reliable ways to determine the presence of these organisms in various samples and (ii) hides and faeces are the main reservoirs of BPS clostridia in the abattoirs. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper provides useful information to detect BPS organisms, as well as to develop a science-based control strategy of the problem.

Changes in Escherichia coli O157:H7 numbers during holding on excised lean, fascia and fat beef surfaces at different temperatures.

AIM: To investigate changes in Escherichia coli O157:H7 numbers on excised beef carcass surfaces over 72 h at different temperatures. METHODS AND RESULTS: Excised lean meat, fascia and fat were inoculated with E. coli O157:H7 and held in an environmental chamber for 72 h, at air speed 0.5 m s(-1), relative humidity (RH) 90%, and temperatures 4, 8 and 12 degrees C. On lean, pathogen counts increased significantly at 12 degrees C. On fascia, significant reductions in counts occurred at 4 and 8 degrees C. Pathogen numbers were significantly reduced on fat at 4, 8 and 12 degrees C (64 h). Counts on fat were significantly less at all temperatures, compared to lean or fascia and surface water activity, a(w), decreased significantly over time on fat at 4 degrees C. Significant decreases in surface pH values were recorded on all meat substrates. CONCLUSIONS: The survival of E. coli O157:H7 varied in relation to the meat substrate and the holding temperature. Reductions in counts on fat surfaces appeared to be related to low surface a(w) values. No relationship between pathogen survival and surface pH was established. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of excised meat pieces in an environmental cabinet offers a more flexible approach to determining the use of different chilling regimes in the production of safe meat.

The potential use of chilling to control the growth of Enterobacteriaceae on porcine carcasses and the incidence of E. coli O157:H7 in pigs.

AIMS: To (i) monitor the presence of Enterobacteriaceae as indicators of faecal contamination on pig carcasses, (ii) examine the potential use of chilling as a critical control point (CCP) and establish its influence on pig carcass categorization by Decision 471/EC and (iii) determine the incidence of E. coli O157:H7 in pigs. METHODS AND RESULTS: Porcine faecal samples and carcass swabs were collected before and after chilling at four Irish pig abattoirs and examined for Enterobacteriaceae and E. coli O157:H7. Chilling generally reduced Enterobacteriaceae counts on carcasses, but increases were also observed, particularly in one abattoir. E. coli O157:H7 was absent from carcasses before chilling, present on 0.21% after chilling and was recovered from 0.63% of faecal samples. All of the isolates were found to contain virulence genes associated with clinical illness in humans. CONCLUSIONS: The data show that overall chilling had the capacity to reduce the numbers of carcasses positive for the presence of Enterobacteriaceae. SIGNIFICANCE AND IMPACT OF STUDY: The influence of chilling on the categorization of pig carcasses suggests that it has the potential to improve the numbers of acceptable carcasses and the process could be used as a CCP within a HACCP plan.

The survival of Salmonella enterica serovar Typhimurium DT104 and total viable counts on beef surfaces at different relative humidities and temperatures.

AIMS: The aim of this study was to investigate changes in Salmonella and total viable count (TVC) survival on beef carcass surfaces stored for 72 h under different combinations of relative humidity (i.e. RH 75% or 96%) and temperature (5 degrees C or 10 degrees C). METHODS AND RESULTS: The influence of low water activity (a(w)) and temperature on the survival and growth of Salmonella enterica serovar Typhimurium DT104 and the aerobic mesophilic flora on meat pieces from different sites on beef carcasses was investigated, under controlled conditions (75% or 96% RH; 5 or 10 degrees C) in an environmental cabinet. Salmonella counts declined during storage at low a(w) (75% RH) conditions at 5 degrees C or 10 degrees C. Salmonella counts increased during storage at high a(w) (96% RH) at 10 degrees C only. At 5 degrees C, TVCs increased during storage at high a(w), but not at low a(w). TVCs increased on all samples from carcasses stored at high or low a(w) at 10 degrees C, except those samples taken from areas of surface fat. CONCLUSIONS: This suggests that substrate composition dictates growth rates under low a(w) conditions. The results are discussed in terms of the possible protective effects of substrate osmolyte accumulation in bacterial survival and/or growth. SIGNIFICANCE AND IMPACT OF THE STUDY: The data obtained in this study provides useful insights on the influence of a(w) and temperature on pathogen survival on meat surfaces at chill temperature.

Molecular characterization of Irish E. coli O157:H7 isolates of human, bovine, ovine and porcine origin.

AIMS: To determine the degree of relatedness between isolates of Escherichia coli O157:H7 of human, bovine, ovine and porcine origin. METHODS AND RESULTS: Escherichia coli O157:H7 isolates were compared using (i) PFGE XbaI patterns, (ii) PCR profiles of virulence genes and (iii) the DNA sequences of genes reported to play a role in pathogenicity. The 77 E. coli O157:H7 isolates demonstrated 49 different PFGE patterns of which, eight were common to multiple isolates, and the remaining 41 were distinct. Isolates of different origin did not correlate, except for one cluster consisting of two human and two beef isolates. The majority of animal isolates had the same PCR profiles of virulence genes as those isolated from clinical patients. Single nucleotide polymorphisms (SNPs) were identified in the sequence of a 255-bp region of the vtx2 subunit A gene. CONCLUSIONS: Six SNPs were detected in the vtx2A gene, defining four different haplotypes. One nonsynonymous substitution encoded for an amino acid change from glutamic to aspartic acid. SIGNIFICANCE AND IMPACT OF THE STUDY: Results indicate that although E. coli O157:H7 isolates of differing origin were distinct by PFGE, the DNA sequences of the main virulence genes associated with human clinical illness were conserved.

Survival of Escherichia coli O157:H7 and non-pathogenic E. coli on irradiated and non-irradiated beef surfaces.

This study examined changes in numbers of pathogenic (PEC) and non-pathogenic (NPEC) Escherichia coli during storage at 10°C on the surfaces of irradiated (IR) and non-irradiated (NIR) meat pieces excised from the neck, brisket and rump of beef carcasses and in Brain Heart Infusion Broth (BHI) and Maximum Recovery Diluent (MRD). On irradiated meat pieces, there were significant differences between mean PEC and NPEC counts at all sites. Differences in counts were also observed between IR and NIR surfaces and among the three meat sites for both E. coli types. These differences occurred only on IR samples, suggesting that the irradiation associated reductions in normal beef surface flora influenced survival of both E. coli types. PEC and NPEC counts increased during storage in BHI, but only NPEC counts increased in MRD. The results of this study highlight the impact of meat surface type and the presence/absence of the normal beef carcass surface flora on E. coli survival and/or growth during meat storage. Such previously unreported effects, and their precise mechanisms, have direct implications in the development and application of accurate models for the prediction of the safety and shelf life of stored meat.

Assessing the hygiene of pig carcasses using whole-body carcass swabs compared with the four-site method in EC Decision 471.

An investigation was carried out in a pig abattoir to determine the microbiological status of carcasses being produced after slaughter and dressing. The carcasses were sampled in accordance with EC Decision 471 in relation to the application of hazard analysis critical control point (HACCP) criteria to the slaughter of animals. In this regard, four sites on the animals were examined on five consecutive carcasses during each of 10 visits for the presence of total viable counts and Enterobacteriaceae. A comparison of the EC four-site method, with a whole-body swab technique, as a means of measuring carcass contamination found that the two methods gave significantly different results for both groups of organisms. A comparison of the mean of the individual data from the four sites with the data from the pooled samples revealed that there was a poor relationship between the two. Samples may be taken by excision or swabbing and allocated to three categories of process control, which, in turn, are based on microbiological criteria that are different, depending on whether sampling is by excision or swabbing. The influence of these changes in microbiological criteria is discussed in relation to the categorization of samples as acceptable, marginal, or unacceptable and the influence this has on process control. Finally, the proposed introduction of Salmonella as a safety indicator in the EC HACCP system is discussed.

Distribution of airborne microorganisms in commercial pork slaughter processes.

The objective of this research was to determine the prevalence and distribution of airborne bacterial contamination, with particular reference to Escherichia coli and Salmonella, at a number of stages in a pork slaughtering plant. Air samples (impaction and sedimentation) were recovered from seven locations before and during operations in a commercial pork processing plant. Aerobic mesophilic bacteria, E. coli counts and the incidence of Salmonella in the air were determined. Most sample locations which provided high impaction counts also provided high sedimentation counts. Before commencement of operations, there were no significant differences in aerobic mesophilic bacteria obtained from the sample locations. However, within 2 h of the commencement of operations, aerobic mesophilic bacteria in the wet room (3.14 log10 cfu/m3) were significantly higher (P < 0.05) than those in the clean room (2.66 log10 cfu/m3) and chiller (2.34 log10 cfu/m3). By the afternoon, similar aerobic mesophilic bacteria counts were recovered in the wet and clean rooms, although counts in both of these areas were significantly higher (P > 0.05) than in the chiller. In general there were no significant differences in E. coli counts between rooms (wet room, clean room and chiller) and these did not increase during the production day. Salmonella were detected at the locations of the dehairing and evisceration operations. Aerobic mesophilic bacteria in the air within the abattoir increased as production proceeded. In addition the air within the abattoir contained organisms such as Salmonella and E. coli. Positive correlations (P < 0.05-P < 0.001) between impaction and sedimentation samples were found suggesting that air may be an important source of carcass contamination.

The characterisation of E. coli O157:H7 isolates from cattle faeces and feedlot environment using PFGE.

The objectives of this study were to investigate the diversity of Escherichia coli O157:H7 isolates obtained over a 3-month period from a cattle feedlot in order to assess the relationship between environmental and faecal isolates and to determine the pattern of transmission of E. coli O157:H7 between groups of cattle. Faecal samples were obtained from cattle housed in four adjacent feedlot pens at monthly intervals, with environmental pen samples collected simultaneously. All E. coli O157:H7 isolates obtained were examined by pulsed field gel electrophoresis (PFGE), polymerase chain reaction (PCR) to detect eaeA, ehxA, stx1 and stx2 genes and antibiotic sensitivity profiling. Ten isolates were subjected to acid shock to imitate conditions in the acidic cattle abomasum and assess the effect on PFGE profiles. E. coli O157:H7 was isolated from 69 faecal samples and 26 environmental samples. All isolates (n=95) carried the genes for eaeA, ehxA and stx2 and were sensitive to all antibiotics tested. The PFGE profiles of all isolates differed by no more than two bands and clustered within 80% similarity following dendrogram analysis. Acid shock had no effect on the subsequent PFGE patterns. A total of 8.7% (6/69) of cattle were shedding E. coli O157:H7 in the first month with faecal shedding increasing to 52% (36/69) by the third month of the study. A single isolate of E. coli O157:H7 may be passed rapidly through cattle pens, with the environment acting as a significant reservoir for transmission. PFGE is a useful tool for tracking the direct and indirect transmission of E. coli O157:H7 isolates on the farm.

A comparison of the survival in feces and water of Escherichia coli O157:H7 grown under laboratory conditions or obtained from cattle feces.

Escherichia coli O157:H7 is an important foodborne pathogen that can cause hemorrhagic colitis and hemolytic uremic syndrome. Cattle feces and fecally contaminated water are important in the transmission of this organism on the farm. In this study, the survival of E. coli O157:H7 in feces and water was compared following passage through the animal digestive tract or preparation in the laboratory. Feces were collected from steers before and after oral inoculation with a marked strain of E. coli O157:H7. Fecal samples collected before cattle inoculation were subsequently inoculated with the marked strain of E. coli O157:H7 prepared in the laboratory. Subsamples were taken from both animal and laboratory-inoculated feces to inoculate 5-liter volumes of water. E. coli O157:H7 in feces survived up to 97 days, and survival was not affected by the method used to prepare the inoculating strain. E. coli O157:H7 survived up to 109 days in water, and the bacteria collected from inoculated cattle were detected up to 10 weeks longer than the laboratory-prepared culture. This study suggests that pathogen survival in low-nutrient conditions may be enhanced by passage through the gastrointestinal tract.

Characterisation of E. coli O157 isolates from bovine hide and beef trimming in Irish abattoirs by pulsed field gel electrophoresis.

Escherichia coli O157 isolates from bovine hide (n=117) and beef trimmings (n=32) from a single abattoir were examined by pulsed field gel electrophoresis (PFGE). Using BioNumerics software, dendrograms of isolates from each sample type (i.e. hide and beef trimming) were produced. In assessing the genetic relatedness of isolates, a similarity criterion of 80% was applied. The 117 E. coli O157 hide isolates were grouped into 14 clusters, comprising of 109 different PFGE profiles. Of the 109 different PFGE profiles, 8 were common to multiple isolates (i.e. shared 100% similarity by PFGE). The 32 E. coli O157 beef trimming isolates produced 28 different PFGE profiles and 2 clusters. Of the 28 PFGE profiles, 2 were common to multiple isolates and the remaining 26 were distinct. On a number of sampling occasions, isolates displaying identical PFGE patterns were recovered from multiple isolates collected from a single sample type (i.e. hides or trimmings), suggesting cross contamination from contaminated hides/animals to uncontaminated hides/animals and from contaminated beef trimmings to uncontaminated beef trimmings during abattoir operations.

The prevalence and characterisation of Cryptosporidium spp. in beef abattoir water supplies.

The prevalence of Cryptosporidium spp. in 50 l samples of water used to wash beef carcasses at (a) an abattoir with a borehole water (BH) supply (n = 46) and (b) an abattoir with a river water (RW) supply (n = 48) was determined. In addition, a 100 l water sample and post-wash carcass samples (n = 24) were collected from the RW supply on a single day in July. Cryptosporidium spp. was detected in 0% and 26.1% of samples from the BH and RW supply abattoirs, respectively, with oocyst concentrations ranging from 0.02 to 8.6/l. Cryptosporidium spp. was not isolated from post-wash beef carcasses, while it was detected in water samples from that day at a concentration of 0.06 oocysts/l. The species of 3/5 isolates were identified as C. parvum, and the remaining were C. andersoni. This study has demonstrated that water used to wash beef carcasses can be contaminated with Cryptosporidium of human health importance and is a potential source of carcass contamination.

Prevalence and numbers of Escherichia coli O157 on bovine hides at a beef slaughter plant.

In this study, we investigated the prevalence and numbers of Escherichia coli O157 on bovine hides. Samples (n = 1,500) were collected over a 17-month period (30 samples per week) by sponge swabbing approximately 122-cm2 areas of the bovine rump of slaughtered cattle at an early stage of carcass processing (first legging). Sponge samples (n = 1,500) were stomached in buffered peptone water supplemented with novobiocin, directly plated on sorbitol MacConkey with Cefixime tellurite (SMAC-CT), enriched for 24 h, extracted by immunomagnetic separation, and plated onto SMAC-CT agar. Presumptive E. coli O157 colonies from SMAC-CT plates were confirmed by PCR for the presence of eaeA, hlyA, fliCh7, vt1, vt2, and portions of the rfb (O-antigen encoding) region of E. coli O157. Overall, E. coli O157 was recovered from 109 samples (7.3%) at concentrations ranging from less than 0.13 to 4.24 log CFU/100 cm2. PCR analysis revealed a wide diversity of genetic profiles among recovered isolates of verocytotoxigenic E. coli. Of the isolates recovered, 99 of 109 contained the attaching and effacing gene (eaeA) and the hemolysin gene (hlyA), and 78 of 109 had the flagellar H7 antigen-encoding gene (fliCh7). Only 6 of 109 isolates contained both verotoxin-producing genes (vt1 and vt2); 91 of 109 contained the vt2 gene only, whereas 1 of 109 contained the vt1 gene only. The remaining 11 of 109 contained neither vt1 nor vt2.

Prevalence and characterisation of Cryptosporidium species in cattle faeces and on beef carcases at slaughter.

Cattle are known reservoirs and asymptomatic excretors of Cryptosporidium, a protozoan parasite that causes severe and protracted diarrhoea in people. The incidence of Cryptosporidium was investigated in 288 matched samples taken from beef carcases of 1 g samples of faeces retrieved immediately after de-legging, 25 cm2 samples of beef excised from the rump of uneviscerated carcases, and 25 cm2 samples of beef excised from the brisket area of eviscerated carcases. Cryptosporidium species were detected in 21 of the faecal samples after salt flotation and immunofluorescent microscopy. The species isolated from the positive samples were identified by restriction fragment length polymorphism and PCR as Cryptosporidium andersoni (54.5 per cent) and Cryptosporidium parvum genotype 2 (45.5 per cent). In the faecal samples, there was a significantly higher prevalence of the parasite in samples taken in summer (May to July) and winter (November to January) than in spring or autumn. No Cryptosporidium species were recovered from any of the beef samples.