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Simulating Escherichia coli O157:H7 transmission to assess effectiveness of interventions in Dutch dairy-beef slaughterhouses.

Beef contamination with Escherichia coli O157:H7 (VTEC) is an important food-safety issue. To investigate the effectiveness of interventions against VTEC in Dutch beef industrial slaughterhouses that slaughter 500 dairy cattle per day, a Monte Carlo simulation model was built. We examined seven carcass-antimicrobial interventions, namely: hot-water wash, lactic-acid rinse, trim, steam-vacuum, steam-pasteurization, hide-wash with ethanol and gamma irradiation, and their combinations. The estimated daily prevalence of contaminated beef-carcass quarters as the output of the model was 9.2%. Contaminated was defined as containing one or more CFU on the surface of a carcass quarter at the end of the quartering stage. Single interventions (except irradiation) could reduce the prevalence to from 6.2% to 1.7%, whereas the combination of interventions could lower it to from 1.2% to 0.1%. The most powerful intervention was irradiation, which could reduce the prevalence to <0.1%. The results of this study indicate that application of single interventions might be useful, although not sufficient. Hence, a combination of interventions along the slaughter process is the more promising approach to reduce the prevalence of contaminated beef quarters.

Abattoirs↗

High prevalence of multiple resistance to antibiotics in Salmonella serovars isolated from a poultry slaughterhouse in Spain.

Salmonellosis is a major foodborne infection in Spain, and strains that are resistant to a great variety of antibiotics have become a major public health concern. The aim of this study was to determine the level of antibiotic resistance in 133 Salmonella isolates obtained from a poultry slaughterhouse in Zaragoza (NE Spain). Antimicrobial resistance testing was performed by disk diffusion method using 19 antibiotics. Results were interpreted following the NCCLS criteria. Overall, the highest percentage of resistance was found to the following antimicrobial agents: sulfadiazine (96.2%), neomycin (53.4%), tetracycline (21.8%), and streptomycin (11.3%). All isolates were found to be resistant to one or more of the antibiotics tested. Multiple resistance was observed in 87 strains (65.4%). We found 23 different patterns of resistance in Salmonella Enteritidis. Resistance to sulfadiazine was the most common single resistance. The most frequent patterns of multiresistant strains were neomycin+sulfadiazine and neomycin+tetracycline+sulfadiazine. S. 4,5,12:b: showed the highest percentages of resistance to the tested drugs, with five different resistance patterns found. Ampicillin+chloramphenicol+streptomycin+sulphonamides+tetracycline (ACSSuT) resistance pattern, commonly associated with S. Typhimurium DT 104, was not detected in strains of the same phage type from broilers. The appearance of substantial multiresistance in foodborne Salmonella isolates suggests the need for more prudent use of antibiotics by farmers, veterinarians, and physicians.

Abattoirs↗

Isolation of Cryptosporidium andersoni Kawatabi type in a slaughterhouse in the northern island of Japan.

Fecal samples were collected from 325 adult cattle and 108 pigs in a slaughterhouse in Hokkaido, the northern island of Japan. Five adult cattle were found to be positive for oocysts of Cryptopsoridium (1.5%). The oocysts were morphologically similar to those of Cryptosporidium andersoni. The partial sequence of the 18S rRNA gene of the isolate was 100% identical with that of the C. andersoni Kawatabi strain. SCID mice were infected after oral administration. Based on the morphology of the oocysts, the sequence of the 18S rRNA gene and the infectivity to SCID mice, the isolate was concluded to be of the same type as the C. andersoni Kawatabi strain that has been isolated in Honshu, the main island of Japan.

Abattoirs↗

Chemical and biological characterization of slaughterhouse wastes compost.

The chemical and biological properties of compost made from yard trimmings (YT) composted alone or mixed with slaughterhouse wastes (SHW) were evaluated in seven phases. Mixtures were weighed in a 2:1 proportion (YT:SHW) and placed in composting bins (0.91 m2). Temperature was recorded to determine the time (d) needed to reach the first (1HC) and second heat cycles (2HC). Composting characteristics were measured at 0 d, at the peak of the 1HC and 2HC, and at maturation (0, 20, 50 and 70 d). During 1HC, bacterial isolates were cultivated in both treatments and identified using the Biolog System. Chemical composition was statistically analyzed using a 2 (layers of SHW)x7 (composting phases) factorial arrangement of treatments with the ANOVA procedure of SAS. The pH was neutral for YT and ranged from 7.41 to 6.82 for SHW throughout the process. There was a decrease in organic matter (OM) and carbon (C), and a relative increase in nitrogen (N) in both treatments. At 70 d of maturation, C:N values were similar between treatments, but lower (P>0.05) than the initial values. Final N concentration was higher (P>0.05) for the treatment with SHW. Only the SHW treatment exhibited thermophilic temperatures. At the 1HC in both treatments, different populations of bacteria responsible for the breakdown of OM were identified showing an active heterogeneous population. The presence of pathogenic microorganisms was not detected in treatments containing SHW.

Abattoirs↗

Integrated anaerobic-aerobic fixed-film reactor for slaughterhouse wastewater treatment.

An integrated anaerobic-aerobic fixed-film pilot-scale reactor with arranged media was fed during 166 days with slaughterhouse wastewater. Operation temperature was 25 degrees C and the anaerobic-aerobic volume ratio was decreased from 4:1 to 3:2 and finally to 2:3. Overall organic matter removal efficiencies of 93% were achieved for an average organic loading rate of 0.77 kg COD/m3 d, and nitrogen removal efficiencies of 67% were achieved for nitrogen loading rates of 0.084 kg N/m3 d. The high internal recirculation associated to the air-lift effect linked to the aeration of a part of the reactor section caused high mixing between the anaerobic and aerobic zones, so that most organic matter was removed aerobically. The nitrification process achieved an efficiency of 91% for nitrogen loads of 0.15 kg N/m3 d when the anaerobic-aerobic volume ratio was 2:3 and was limited by dissolved oxygen concentration below 3 mg/l. The influence of the heterotrophic biomass growing in the outer biofilm was checked. Denitrification only implied the 12-34% of the total nitrogen removal and was limited by dissolved oxygen concentration in the anaerobic zone above 0.5 mg/l caused by the mixing regime. Most removed nitrogen was employed in synthesis of heterotrophic bacteria.

Bacteria, Aerobic↗

Element concentrations in the archiacanthocephalan Macracanthorhynchus hirudinaceus compared with those in the porcine definitive host from a slaughterhouse in La Paz, Bolivia.

Concentrations of lead and cadmium, determined by electrothermal atomic absorption spectrometry, and concentrations of the elements barium, cadmium, copper, iron, magnesium, manganese, nickel, lead, selenium and strontium, determined by inductively coupled plasma mass spectrometry, in the acanthocephalan Macracanthorhynchus hirudinaceus and its porcine final host, sampled at a slaughterhouse in La Paz, Bolivia, were compared. Inductively coupled plasma mass spectrometry analysis revealed that most of the elements were found at higher concentrations in the acanthocephalan than in different tissues of its host. The bioconcentration of elements in M. hirudinaceus compared with the host intestine, listed in order of decreasing values, was as follows: Cd > Pb > Ni > Sr = Cu > Mg > Se > Fe = Mn = Ba. Analysis by electrothermal atomic absorption spectrometry showed that M. hirudinaceus contained 85, 85, 56 and 24 times higher lead levels compared with hosts muscle, liver, kidney and intestine, respectively. The mean cadmium concentration of the parasite was 32 times higher than that of the liver and five times higher compared with porcine kidney. The metal distribution within the body of M. hirudinaceus was as follows: cement gland > testes > lemnisci > eggs = tegument for lead and lemnisci > testes > cement gland > tegument > eggs for cadmium. Therefore, the hypothesis that parasites excrete toxic metals with the shells of their eggs seems not to be valid for M. hirudinaceus. It is concluded, that not only eoacanthocephalans and palaeacanthocephalans parasitising fish, but also archiacanthocephalans from mammalian hosts, are able to bioaccumulate metals.

Abattoirs↗

Semi-continuous anaerobic digestion of solid poultry slaughterhouse waste: effect of hydraulic retention time and loading.

We studied the effect of hydraulic retention time (HRT) and loading on anaerobic digestion of poultry slaughterhouse wastes, using semi-continuously fed, laboratory-scale digesters at 31 degrees C. The effect on process performance was highly significant: Anaerobic digestion appeared feasible with a loading of up to 0.8 kg volatile solids (VS)/m3 d and an HRT of 50-100 days. The specific methane yield was high, from 0.52 to 0.55 m3/kg VS(added). On the other hand, at a higher loading, in the range from 1.0 to 2.1 kg VS/m3 d, and a shorter HRT, in the range from 25 to 13 days, the process appeared inhibited and/or overloaded, as indicated by the accumulation of volatile fatty acids and long-chain fatty acids and the decline in the methane yield. However, the inhibition was reversible. The nitrogen in the feed, ca. 7.8% of total solids (TS), was organic nitrogen with little ammonia present, whereas in the digested material ammonia accounted for 52-67% (up to 3.8 g/l) of total nitrogen. The TS and VS removals amounted to 76% and 64%, respectively. Our results show that on a continuous basis under the studied conditions and with a loading of up to 0.8 kg VS/m3 d metric ton (wet weight) of the studied waste mixture could yield up to 140 m3 of methane.

Abattoirs↗

Microscopic observation of particle reduction in slaughterhouse wastewater by coagulation-flocculation using ferric sulphate as coagulant and different coagulant aids.

A coagulation-flocculation treatment has been applied to a slaughterhouse liquid effluent, using ferric sulphate as coagulant and activated silica, powdered activated carbon, cationic polyacrylamide, polyvinyl alcohol, polyacrylic acid and anionic polyacrylamide as coagulant aids in order to improve the settling time. Once the optimal conditions had been established (speed and time of stirring during flocculation step, pH, coagulant and coagulant aid doses), the efficiency of the coagulation-flocculation process was studied by comparing the particle size distribution before and after the addition of the coagulant. When ferric sulphate was used without the coagulant aids, particle removal efficiency varied with size, although overall efficiency was quite substantial (87%). The use of coagulant aids improved the removal efficiency, except in the case of activated silica, when it fell to 78%. In all other cases removal efficiency values between 93% (for polyvinyl alcohol) and 99% (for anionic polyacrylamide) were reached.

Abattoirs↗

Prevalence of vancomycin-resistant Enterococcus faecium (VREF) in pig faeces from slaughterhouses in Spain.

A study to estimate the prevalence of vancomycin-resistant Enterococcus faecium in faecal samples from pigs at slaughterhouses in Spain was carried out between November 1998 and January 1999 with 900 samples taken from four abattoirs representing 9.7% of all pig slaughtered in 1998. Using a selective enrichment broth with vancomycin (8microg/ml), 64 samples (7.1%; 95% CI: 5.5, 9.0%) had E. faecium vancomycin-resistant strains that showed minimal inhibitory concentrations of 256microg/ml (62 strains) and 512microg/ml (two strains). Results by farm showed that 43 of the 240 pig farms represented in the sampling had at least one faecal sample with vancomycin-resistant E. faecium.

Abattoirs↗

Campylobacter jejuni contamination on broiler carcasses of C. jejuni-negative flocks during processing in a Japanese slaughterhouse.

At a slaughterhouse, we determined the Campylobacter jejuni status of each broiler flock coming from different farms by culturing cecal contents of representative birds, and examined the prevalence of C. jejuni on carcasses during processing. Then, C. jejuni isolates from the carcasses were typed with RAPD PCR and compared with those from the cecal contents of a C. jejuni-positive flock. C. jejuni was not isolated from the carcasses of C. jejuni-negative flocks processed before the C. jejuni-positive flock, and C. jejuni was isolated from the carcasses of C. jejuni-negative flocks processed after the C. jejuni-positive flock. RAPD PCR type of the isolates from the carcasses of C. jejuni-negative flocks was the same as those from the cecal contents of the previously processed C. jejuni-positive flock. Our results suggest that the carcasses of the C. jejuni-negative flocks were contaminated with C. jejuni strains originating from the intestines of the previously processed C. jejuni-positive flock.

Abattoirs↗

Frequency of Shiga toxin-producing Escherichia coli in cattle at a breeding farm and at a slaughterhouse in Japan.

To investigate the frequency of Shiga toxin-producing Escherichia coli (STEC) infected calves at a breeding farm and cattle at a slaughterhouse in Tohoku area of Japan, the polymerase chain reaction (PCR) was used for detection of genes for Shiga toxin(s). The fecal samples from a total of 204 calves and 306 cattle were examined. The prevalence rates in calves less than 2 months of age, cattle 2-8 months of age, and adults greater than 1 year of age were 39.4, 78.9, and 40.8%, respectively. Detection frequency of STEC in the fecal specimens from calves aged 0-8 months was not different among the breeds of cattle (Holstein: H, Japanese black cattle: B, and F1: HxB). On the other hand, for calves over 12 months of age, the frequency of STEC in Japanese black cattle and F1 were significantly higher than in Holstein cattle. Serogroups of STEC usually identified in human cases of food poisoning (O157, O26, and O111) were not frequently found in the feces of the cattle.

Abattoirs↗

Surveillance of antimicrobial resistance in Escherichia coli strains isolated from pigs at Spanish slaughterhouses.

Antimicrobial resistance can make the efficient treatment of bacterial infections in humans and animals more difficult. Antimicrobial use in food animals may be one of the factors contributing to resistance. The Spanish surveillance network VAV has established a baseline of antimicrobial resistance in Escherichia coli strains from healthy pigs. Minimum inhibitory concentration and patterns of resistance to antimicrobials used in animals and humans were determined for 205 faecal strains isolated in a sampling frame of four slaughterhouses in Spain from 220 pigs in 1998. Higher levels of resistance were seen against antimicrobial agents authorised for use in food animals especially tetracycline, sulphonamides, trimethoprim and amoxycillin. All isolates were susceptible to antimicrobials employed mainly in humans such as ceftazidime, cefotaxime, imipenem, aztreonam and amikacin.

Abattoirs↗

The effect of temperature on slaughterhouse wastewater treatment in anaerobic sequencing batch reactors.

High strength slaughterhouse wastewater was treated in four 42 l anaerobic sequencing batch reactors (ASBRs) operated at 30 degrees C, 25 degrees C and 20 degrees C. The wastewater contained between 30% and 53% of its chemical oxygen demand (COD) as suspended solids (SS). The ASBRs could easily support volumetric organic loading rates (OLRs) of 4.93, 2.94 and 2.75 kg/m3/d (biomass OLRs of 0.44, 0.42 and 0.14 g/g volatile SS (VSS)/d) at 30 degrees C, 25 degrees C, and 20 degrees C, respectively. At all operating temperatures, the total COD (TCOD) and soluble COD (SCOD) were reduced by over 92%, while average SS removal varied between 80% and 96%. Over the experimental period, 90.8%, 88.7% and 84.2% of the COD removed was transformed into methane at 30 degrees C, 25 degrees C and 20 degrees C, respectively. The decrease in the conversion of the COD removed into methane as operating temperature was lowered, may be partly explained by a lower degradation of influent SS as temperature was reduced. The reactors showed a high average methanogenic activity of 0.37, 0.34 and 0.12 g CH4-COD/gVSS/d (22.4, 12.7 and 11.8 l/d) at 30 degrees C, 25 degrees C and 20 degrees C, respectively. The average methane content in the biogas increased from 74.7% to 78.2% as temperature was lowered from 30 degrees C to 20 degrees C.

Abattoirs↗

Testing of alkaline and enzymatic hydrolysis pretreatments for fat particles in slaughterhouse wastewater.

Four pretreatments to hydrolyse and/or reduce the size of fat particles in slaughterhouse wastewater (SHW) were tested: sodium hydroxide and three lipases of plant, bacterial and animal (pancreatic) origin. Hydrolysing agents and SHW containing between 2.5 and 3 g/l of fat particles were mixed at room temperature for 4 h. Additions of 5-400 meq NaOH/l did not increase soluble COD (SCOD) in SHW, but the average particle size was reduced to 73% +/- 7% of the initial average particle size (D(in)) at NaOH concentrations ranging from 150 to 300 meq/l. Pretreatment with pancreatic lipase PL-250 reduced the average particle size to a maximum of 60% +/- 3% of D(in). As D(in) was decreased from 359 to 68 microns, the enzyme concentration required to obtain the maximum particle size reduction increased from 200 to 1000 mg/l. A 4-h pretreatment with PL-250 also increased the free long-chain fatty acid (LCFA) concentration to a maximum of 15.5 mg/l, indicating some solubilization of the pork fat particles in SHW. SCOD was not significantly increased by the pretreatment, but SCOD was not found to be a good indicator of enzymatic lipolysis because of enzyme adsorption on the fat particle surface. Pancreatic lipase appeared more efficient with beef fat than pork fat, possibly because beef fat contains less polyunsaturated fatty acids than pork fat. The bacterial lipase LG-1000 was also efficient in reducing average fat particle size, but high doses (> 1000 mg/l) were required to obtain a significant reduction after 4 h of pretreatment. SCOD was not increased by pretreatment with LG-1000. No particle size reduction or changes in SCOD were noted after 4 h of pretreatment with the plant lipase EcoSystem Plus. It was concluded that PL-250 was the best pretreatment to hydrolyse fat particles in SHW. However, its impact on the efficiency of a downstream anaerobic digestion process remains to be tested.

Abattoirs↗

Anaerobically digested poultry slaughterhouse wastes as fertiliser in agriculture.

Chemical and physical analysis, 27-d plant growth assays with carrot (Daucus carota) and Chinese cabbage (Brassica campestris var. chinensis), and 5-d phytotoxicity assays with Chinese cabbage and perennial ryegrass (Lolium perenne) were used to investigate the suitability of anaerobically digested poultry slaughterhouse waste for fertiliser in agriculture and the effect of aerobic post-treatment on the properties of the digested material. The digested material appeared to be rich in nitrogen. In 27-d assays with digested material as nitrogen source, carrots grew almost as well as those fertilised with a commercial mineral fertiliser used as reference, whereas, the growth of Chinese cabbage was inhibited. In further 5-d phytotoxicity assays, the digested material inhibited the germination and root growth of ryegrass and Chinese cabbage, apparently because of organic acids present in it. Aerobic post-treatment of the material reduced its phytotoxicity but, probably due to the volatilisation of ammonia, resulted in loss of nitrogen.

Abattoirs↗

Isolation and characterization of two glycerol-fermenting clostridial strains from a pilot scale anaerobic digester treating high lipid-content slaughterhouse waste.

Two obligately anaerobic bacterial strains were isolated from the contents of a pilot scale, anaerobic digester treating slaughterhouse waste with a high protein and lipid content. The isolates, LIP1 and MW8, were characterized as spore-forming, Gram-positive rods, capable of fermenting glycerol. Isolate LIP1 was also observed to be lipolytic and was able to hydrolyse tallow and olive oil. Both isolates grew optimally at 37 degrees C and formed either acetate and formate (LIP1), or acetate and butyrate (MW8), as major glycerol fermentation products. Both isolates produced ethanol as the major reduced fermentation end-product. Neither MW8 nor LIP1 had growth and metabolism inhibited by the addition of stearic acid at concentrations normally considered bactericidal. Analysis of the 16S rRNA gene sequences, in conjunction with the phenotypic data, confirmed that the isolates are members of the genus Clostridium (sensu lato), clustering with species of clostridial clusters I (MW8) and XIVa (LIP1).

Abattoirs↗

Cross-contamination with Salmonella on a broiler slaughterhouse line demonstrated by use of epidemiological markers.

AIMS: To investigate contamination of surfaces on a poultry slaughter line from infected poultry and subsequent cross-contamination of non-infected poultry. METHODS AND RESULTS: A broiler slaughterhouse was investigated for the presence of Salmonella on 17 defined points over two 1-week periods. Flocks supplied to slaughter and neck skin samples from processed chicken were likewise investigated. Salmonella was detected in 10 out of 18 flocks at ante-mortem inspection, while seven flocks tested positive in the finished products. Equipment at all but one control point at the slaughter line tested positive at least once during the study. The chicken receiving area was the most contaminated. By comparison of typing results from serotyping, plasmid profile typing and phage typing, direct evidence for cross-contamination with Salm. serotype Typhimurium, Salm. Serotype 4.12:b:- and Salm. serotype Virchow on the slaughter line was obtained for four of the flocks. The cleaning procedure in place did not remove all Salmonella from the contaminated areas. CONCLUSIONS: Evidence for contamination of equipment on a slaughter line and subsequent cross-contamination to non-infected chicken was provided by typing methods. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has provided detailed information on cross-contamination on a slaughter line by the use of phage typing and plasmid profiling. The study stresses the importance of controlling Salmonella in the primary production, as contamination of the equipment on the slaughter line will act as a vehicle to contaminate finished products. Cleaning procedures on slaughter lines cannot be expected to control this problem with the current equipment.

Abattoirs↗

Molecular characterization of the diversity of Campylobacter spp. isolates collected from a poultry slaughterhouse: analysis of cross-contamination.

Investigations of a free-range broiler flock during the rearing period and at the slaughterhouse by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the flagellin (flaA) gene (flaA typing) have shown that poultry carcasses are contaminated by Campylobacter spp. strains which were previously present in the poultry faces. Moreover, the investigation of the previous and the following batches in the processing plant using flaA typing have shown that cross-contamination between batches coming from different flocks occurs and is also a risk factor for the presence of Campylobacter spp. on poultry carcasses.

Abattoirs↗