The wastewater of livestock slaughterhouses is considered a source of antimicrobial-resistant bacteria with clinical relevance and may thus be important for their dissemination into the environment. To get an overview of their occurrence and characteristics, we investigated process water (n = 50) from delivery and unclean areas as well as wastewater (n = 32) from the in-house wastewater treatment plants (WWTPs) of two German poultry slaughterhouses (slaughterhouses S1 and S2). The samples were screened for ESKAPE bacteria (Enterococcus spp., Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp.) and Escherichia coli. Their antimicrobial resistance phenotypes and the presence of extended-spectrum-β-lactamase (ESBL), carbapenemase, and mobilizable colistin resistance genes were determined. Selected ESKAPE bacteria were epidemiologically classified using different molecular typing techniques. At least one of the target species was detected in 87.5% (n = 28/32) of the wastewater samples and 86.0% (n = 43/50) of the process water samples. The vast majority of the recovered isolates (94.9%, n = 448/472) was represented by E. coli (39.4%), the A. calcoaceticus-A. baumannii (ACB) complex (32.4%), S. aureus (12.3%), and K. pneumoniae (10.8%), which were widely distributed in the delivery and unclean areas of the individual slaughterhouses, including their wastewater effluents. Enterobacter spp., Enterococcus spp., and P. aeruginosa were less abundant and made up 5.1% of the isolates. Phenotypic and genotypic analyses revealed that the recovered isolates exhibited diverse resistance phenotypes and β-lactamase genes. In conclusion, wastewater effluents from the investigated poultry slaughterhouses exhibited clinically relevant bacteria (E. coli, methicillin-resistant S. aureus, K. pneumoniae, and species of the ACB and Enterobacter cloacae complexes) that contribute to the dissemination of clinically relevant resistances (i.e., blaCTX-M or blaSHV and mcr-1) in the environment. IMPORTANCE Bacteria from livestock may be opportunistic pathogens and carriers of clinically relevant resistance genes, as many antimicrobials are used in both veterinary and human medicine. They may be released into the environment from wastewater treatment plants (WWTPs), which are influenced by wastewater from slaughterhouses, thereby endangering public health. Moreover, process water that accumulates during the slaughtering of poultry is an important reservoir for livestock-associated multidrug-resistant bacteria and may serve as a vector of transmission to occupationally exposed slaughterhouse employees. Mitigation solutions aimed at the reduction of the bacterial discharge into the production water circuit as well as interventions against their further transmission and dissemination need to be elaborated. Furthermore, the efficacy of in-house WWTPs needs to be questioned. Reliable data on the occurrence and diversity of clinically relevant bacteria within the slaughtering production chain and in the WWTP effluents in Germany will help to assess their impact on public and environmental health.

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The holistic approach of “One Health” includes the consideration of possible links between animals, humans, and the environment. In this review, an effort was made to highlight knowledge gaps and various factors that contribute to the transmission of antibiotic-resistant bacteria between these three reservoirs. Due to the broad scope of this topic, we focused on pig production and selected “indicator bacteria”. In this context, the role of the bacteria livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) and extended spectrum beta-lactamases carrying Escherichia coli (ESBL-E) along the pig production was particularly addressed. Hotspots of their prevalence and transmission are, for example, pig stable air for MRSA, or wastewater and manure for ESBL-E, or even humans as vectors in close contact to pigs (farmers and veterinarians). Thus, this review focuses on the biotope “stable environment” where humans and animals are both affected, but also where the end of the food chain is not neglected. We provide basic background information about antibiotics in livestock, MRSA, and ESBL-bacteria. We further present studies (predominantly European studies) in tabular form regarding the risk potentials for the transmission of resistant bacteria for humans, animals, and meat differentiated according to biotopes. However, we cannot guarantee completeness as this was only intended to give a broad superficial overview. We point out sustainable biotope approaches to try to contribute to policy management as critical assessment points in pig housing conditions, environmental care, animal health, and food product safety and quality as well as consumer acceptance have already been defined.

A trial with different concentrations of DL-methionine (DLM) and DL-2-hydroxy-4-methylthiobutanoic acid (DL-HMTBA) in broiler feed was performed to investigate their effect on the meat quality parameters and the shelf life of breast fillet. In total, fillets from 210 male broiler chickens (Ross 308) were tested in seven groups with 30 animals each. Three different concentrations (0.04, 0.12, and 0.32%; on an equimolar basis) of either DLM or DL-HMTBA were added to a basal diet, summing up to seven treatment groups. After slaughter, fillets were packed aerobically and stored at 4°C. The investigated parameters comprised measurements of microbial as well as physicochemical parameters, such as pH, drip loss, cooking loss, and color measurements. Additionally, sensory investigations were conducted and shelf life was calculated. Mean pH values were between 6.1 and 6.4. Drip loss values were low, with mean values below 0.4%. The cooking loss ranged between 22% and 28% on average. The fillets showed a normal initial microbial quality (2.5 log10 cfu/g) and spoilage process with microbial counts of 8.5 log10 cfu/g at the end of storage. The study revealed a significant influence of methionine supplementation on the quality of broiler breast meat in comparison with the basal group. Methionine supplementation led to higher pH values and a higher water binding. Higher concentrations of methionine had a positive influence on the water-holding capacity by lowering the cooking loss. The L∗ value showed a significant negative correlation to the methionine concentration supplemented. No differences in physicochemical as well as sensory parameters could be detected between both methionine sources. The fillets showed a normal sensory spoilage process and a shelf life of 6 d. White striping was positively correlated to fillet weight as well as color values and significantly affected the Purchase Decision, the sensory investigation, and thus the shelf life of the samples.

Due to the high prevalence of colistin-resistant Enterobacteriaceae in poultry and pigs, process waters and wastewater from slaughterhouses were considered as a hotspot for isolates carrying plasmid-encoded, mobilizable colistin resistances (mcr genes). Thus, questions on the effectiveness of wastewater treatment in in-house and municipal wastewater treatment plants (WWTPs) as well as on the diversity of the prevailing isolates, plasmid types, and their transmissibility arise. Process waters and wastewater accruing in the delivery and unclean areas of two poultry and two pig slaughterhouses were screened for the presence of target colistin-resistant bacteria (i.e., Escherichia coli, Klebsiella spp., Enterobacter cloacae complex). In-house and municipal WWTPs (mWWTPs) including receiving waterbodies were investigated as well. Samples taken in the poultry slaughterhouses yielded the highest occurrence of target colistinresistant Enterobacteriaceae (40.2%, 33/82), followed by mWWTPs (25.0%, 9/36) and pig slaughterhouses (14.9%, 10/67). Recovered isolates exhibited various resistance patterns. The resistance rates using epidemiological cut-off values were higher in comparison to those obtained with clinical breakpoints. Noteworthy, MCR-1-producing Klebsiella pneumoniae and E. coli were detected in scalding waters and preflooders of mWWTPs. A total of 70.8% (46/65) of E. coli and 20.6% (7/34) of K. pneumoniae isolates carried mcr-1 on a variety of transferable plasmids with incompatibility groups IncI1, IncHI2, IncX4, IncF, and IncI2 ranging between 30 and 360 kb. The analyzed isolates carrying mcr-1 on transferable plasmids (n = 53) exhibited a broad diversity, as they were assigned to 25 different XbaI profiles. Interestingly, in the majority of colistin-resistant

The aim of this study was to investigate the influence of different methionine sources and concentrations on the quality and spoilage process of broiler meat. The trial was comprised of 7 treatment groups: one basal group (suboptimal in Methionine+Cysteine; i.e., 0.89, 0.74, 0.69% in DM SID Met+Cys in starter, grower, and finisher diets, respectively) and 3 doses (0.10, 0.25, and 0.40%) of either DL-Methionine (DLM) or DL-2-hydroxy-4-methylthio butanoic acid (DL-HMTBA) on an equimolar basis of the DLM-supplemented groups. The broilers were fed the diets for 35 d, then slaughtered and processed. The filets were aerobically packed and stored under temperature controlled conditions at 4°C. Meat quality investigations were comprised of microbial investigations (total viable count and Pseudomonas spp.), pH and drip loss measurements of the filets. The shelf life of the meat samples was determined based on sensory parameters. After slaughtering, all supplemented meat samples showed a high quality, whereby no differences between the 2 methionine sources could be detected for the microbial load, pH, and drip loss. In comparison to the control group, the supplemented samples showed a higher sensory quality, characterized by a fresh smell and fresh red color. Methionine supplementation had a significant influence on meat quality parameters during storage. The microbial load, pH and drip loss of the chicken filets were positively correlated to the methionine concentration. Additionally, the microbial load at the end of storage was positively correlated to pH and drip loss values. Nevertheless, the microbial parameters were in a normal range and the positive correlation to methionine concentration did not affect the sensory shelf life. The mean sensory shelf life of the broiler filets varied between 7 to 9 d. During storage, no difference in the development of sensory parameters was observed between the supplemented groups, while the spoilage process of the basal group occurred slightly faster. In conclusion, methionine concentration, but not methionine source, effected meat quality parameters in breast muscles of broilers.

In calf rearing, the first weeks of life are critical and associated with the highest mortality due to enteric and respiratory diseases. A well-implemented hygiene management can help to protect calves’ health preventively by reducing the load of pathogenic bacteria and interrupting infection chains. The aim of this study was to identify deficiencies in hygiene management of individually housed dairy calves by surveying current practice and examining feeding and housing equipment with different hygiene indicators. On 11 farms, different locations in 2 pens or hutches for individual calf rearing prepared for restocking and 2 feeding buckets per farm, including the inner and outer surfaces of artificial teats, were visually scored for cleanliness and sampled with swabs (housing equipment: n = 167; feeding equipment: n = 120). The sanitation of floors was tested with sock samples (n = 41). A total of 328 samples were analyzed for adenosine triphosphate (ATP) and protein residues, aerobic total viable count (TVC), total coliform count (TCC), Escherichia coli, methicillin-resistant Staphylococcus aureus (MRSA), extended-spectrum β-lactamase-producing bacteria (ESBL), and Salmonella spp. After evaluation of these results, the farmers were informed about the findings and trained on improvement in hygiene management personally. The sampling was repeated after 1 year to detect possible changes in hygiene management. The highest bacterial loads (TVC, TCC, and E. coli) were observed in feeding equipment, especially the inner teat of milk feeding buckets. Environmental samples, primarily the sidewalls and back walls of tested pens and hutches, exhibited the lowest bacterial counts and ATP and protein residues. All samples were negative for MRSA and Salmonella spp. In 10.5% of all samples, ESBL was detected, and in 6.8%, ESBL E. coli was detected, predominately in sock samples, followed by feeding equipment samples. Training in hygiene management showed only limited effects. In conclusion, there is still great potential to improve the implementation of hygiene measures in individual calf housing. In particular, more attention should be paid to the cleaning of feeding buckets and artificial teats, as this is a simple means of interrupting the possible spread of pathogens among calves.

Optimal hygiene management is an essential part of maintaining a high standard of health in conventional pig production systems and for the successful interruption of infection chains. Currently, efficiency assessments on cleaning and disinfection are only performed by visual inspection or are neglected completely. The aim of this study was to evaluate the available methods for on farm monitoring of hygiene, identify critical points in pig pens and use the data obtained for training purposes. In addition to visual inspection by assessing the cleanliness, microbiological swab samples, i.e., aerobic total viable count (TVC), total coliform count, methicillin-resistant Staphylococcus aureus (MRSA), and extended-spectrum β-lactamases-producing bacteria (ESBL), swab samples for ATP as well as protein residues and agar contact plates combined with 3 different culture media, were applied and ranked according to their suitability for livestock farming. Samples were collected on at least 15 critical points from one representative pen on 6 pig fattening farms with various hygiene management practices after cleaning and disinfection. After the first sampling, farmers were trained with their individual results, and sampling was repeated 6 mo after training. Nipple drinkers, feeding tubes (external and inner surface), and troughs (external and inner surface) showed the greatest bacterial loads (TVC: 4.5–6.7 log10 cfu cm-2) and values for ATP and protein residues; therefore, these surfaces could be identified as the most important critical points. Spearman rank correlations (P < 0.01) were found between the different assessment methods, especially for the TVC and ATP (r = 0.82, P < 0.001). For rapid assessment on farms, ATP tests represented an accurate and cost-efficient alternative to microbiological techniques. Training improved cleaning performance as indicated by a lower rating for visual inspection, TVC, ATP, MRSA, and ESBL in the second assessment. The monitoring of cleaning efficiency in pig pens followed by training of the staff constitutes a valuable strategy to limit the spread of infectious diseases and antibiotic-resistant bacteria. Special attention should be paid to the sufficient hygiene of drinkers and feeders.