Dose-response relationship between antimicrobial drugs and livestock-associated MRSA in pig farming.

The farming community can be a vehicle for introduction of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) in hospitals. During 2011-2013, an 18-month longitudinal study aimed at reducing the prevalence of LA-MRSA was conducted on 36 pig farms in the Netherlands. Evaluations every 6 months showed a slight decrease in MRSA prevalence in animals and a stable prevalence in farmers and family members. Antimicrobial use, expressed as defined daily dosages per animal per year, decreased 44% during the study period and was associated with declining MRSA prevalence in pigs. MRSA carriage in animals was substantially higher at farms using cephalosporins. Antimicrobial use remained strongly associated with LA-MRSA in humans regardless of the level of animal contact. A risk factor analysis outlined potential future interventions for LA-MRSA control. These results should encourage animal and public health authorities to maintain their efforts in reducing antimicrobial use in livestock and ask for future controlled intervention studies.

Within-farm dynamics of ESBL/AmpC-producing Escherichia coli in veal calves: a longitudinal approach.

OBJECTIVES: To assess the within-farm dynamics of extended-spectrum ß-lactamase (ESBL)/AmpC-producing Escherichia coli in veal calves. METHODS: Three veal-calf fattening farms were screened. Faecal samples from all calves within a compartment (109-150 per farm) were taken upon arrival on the farm (T0) and after 3, 6, 8 and 10 weeks (T3-T10). ESBL/AmpC genes were characterized by PCR and sequencing. Plasmids were characterized by transformation, PCR-based replicon typing and plasmid multilocus sequence typing (MLST). E. coli genotypes were analysed by MLST. RESULTS: At T0 the prevalence of ESBL/AmpC-producing E. coli ranged from 18% to 26%. These were predominantly isolates carrying blaCTX-M-1 and blaCTX-M-15 genes, located on various plasmids and E. coli sequence types (STs). Farm 1 was negative for ESBL/AmpC-producing E. coli after T0. Farm 2 showed an increase up to 37% at T3, which subsequently decreased gradually to 0% at T10. The presence from T3 to T10 on farm 2 was mainly caused by the clonal spread of a multiresistant E. coli ST57 harbouring blaCTX-M-14 on an IncF F2:A-:B- plasmid. Farm 3 showed a gradual decrease in prevalence to 1.4% at T10, with a relative increase of the identical clonal variant as shown for farm 2. A second clonal variant found in farm 3 was a multiresistant E. coli ST10 harbouring blaCTX-M-14 on an IncK plasmid. CONCLUSIONS: The prevalence of ESBL/AmpC-producing E. coli decreased over time. A clonal spread was observed on farm 2 and farm 3, illustrative of the complex dynamics probably associated with the use of antimicrobials.

Livestock-associated MRSA ST398 carriage in pig slaughterhouse workers related to quantitative environmental exposure.

OBJECTIVES: To assess livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) carriage among workers in pig slaughterhouses and assess associated risk factors, including occupational exposure to LA-MRSA. METHODS: A cross-sectional study in three Dutch pig slaughterhouses was undertaken. Nasal swabs of participants were taken. Nasal swabs and surface wipes, air and glove samples were screened for presence of methicillin-resistant Staphylococcus aureus (MRSA). MRSA was quantitatively determined on gloves and in air samples by culturing and real-time PCR. RESULTS: 11 of 341 (3.2%) participants were identified as nasal MRSA carriers. MRSA-positive workers were predominantly found at the start of the slaughter process. Major risk factors for carriage were working in the lairage and working in the scalding and dehairing area. Most nasal isolates (73%) belonged to the LA-MRSA clone ST398. MRSA ST398-positive environmental samples were found throughout the slaughter process. A clear decrease was seen along the slaughterline in the number of MRSA-positive samples and in the MRSA amount per sample. CONCLUSIONS: This study showed that working in the lairage area or scalding and dehairing area were the major risk factors for MRSA carriage in pig slaughterhouse workers, while the overall prevalence of MRSA carriage is low. Occupational exposure to MRSA decreased along the slaughterline, and the risk of carriage showed a parallel decrease.

High probability of avian influenza virus (H7N7) transmission from poultry to humans active in disease control on infected farms.

An epizootic of avian influenza (H7N7) caused a large number of human infections in The Netherlands in 2003. We used data from this epizootic to estimate infection probabilities for persons involved in disease control on infected farms. Analyses were based on databases containing information on the infected farms, person-visits to these farms, and exposure variables (number of birds present, housing type, poultry type, depopulation method, period during epizootic). Case definition was based on self-reported conjunctivitis and positive response to hemagglutination inhibition assay. A high infection probability was associated with clinical inspection of poultry in the area surrounding infected flocks (7.6%; 95% confidence interval [CI], 1.4%-18.9%) and active culling during depopulation (6.2%; 95% CI, 3.7%-9.6%). Low probabilities were estimated for management of biosecurity (0.0%; 95% CI, 0.0%-1.0%) and cleaning assistance during depopulation (0.0%; 95% CI, 0.0%-9.2%). No significant association was observed between the probability of infection and the exposure variables.

Effectiveness of personal protective equipment and oseltamivir prophylaxis during avian influenza A (H7N7) epidemic, the Netherlands, 2003.

We analyzed the effectiveness of personal protective equipment and oseltamivir use during the 2003 avian influenza A (H7N7) epidemic in the Netherlands by linking databases containing information about farm visits, human infections, and use of oseltamivir and personal protective equipment. Using a stringent case definition, based on self-reported conjunctivitis combined with a positive hemagglutination-inhibition assay, we found that prophylactic treatment with oseltamivir significantly reduced the risk for infection per farm visit from 0.145 (95% confidence interval [CI] 0.078-0.233) to 0.031 (95% CI 0.008-0.073). The protective effect was ≈79% (95% CI 40%-97%). These results are comparable with the reported effect of prophylactic treatment with oseltamivir on human seasonal influenza. No significant protective effect was found for use of respirators or safety glasses, possibly because of limitations of the data.

Back-calculation method shows that within-flock transmission of highly pathogenic avian influenza (H7N7) virus in the Netherlands is not influenced by housing risk factors.

To optimize control of an avian influenza outbreak knowledge of within-flock transmission is needed. This study used field data to estimate the transmission rate parameter (beta) and the influence of risk factors on within-flock transmission of highly pathogenic avian influenza (HPAI) H7N7 virus in the 2003 epidemic in The Netherlands. The estimation is based on back-calculation of daily mortality data to fit a susceptible-infectious-dead format, and these data were analysed with a generalized linear model. This back-calculation method took into account the uncertainty of the length of the latent period, the survival of an infection by some birds and the influence of farm characteristics. After analysing the fit of the different databases created by back-calculation, it could be concluded that an absence of the latency period provided the best fit. The transmission rate parameter (beta) from these field data was estimated at 4.50 per infectious chicken per day (95% CI: 2.68-7.57), which was lower than what was reported from experimental data. In contrast to general belief, none of the studied risk factors (housing system, flock size, species, age of the birds in weeks and date of depopulation) had significant influence on the estimated beta.

Transmission through air as a possible route of exposure for MRSA.

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) is highly prevalent in pigs and veal calves. The environment and air in pig and veal calf barns is often contaminated with LA-MRSA, and can act as a transmission source for humans. This study explores exposure-response relationships between sequence type 398 (ST398) MRSA air exposure level and nasal ST398 MRSA carriage in people working and/or living on farms. Samples and data were used from three longitudinal field studies in pig and veal calf farm populations. Samples consisted of nasal swabs from the human participants and electrostatic dust fall collectors capturing airborne settled dust in barns. In both multivariate and mutually adjusted analyses, a strong association was found between nasal ST398 MRSA carriage in people working in the barns for >20 h per week and MRSA air levels. In people working in the barns < 20 h per week there was a strong association between nasal carriage and number of working hours. Exposure to ST398 MRSA in barn air seems to be an important determinant for nasal carriage, especially in the highly exposed group of farmers, next to duration of contact with animals. Intervention measures should therefore probably also target reduction of ST398 MRSA air levels.

Effects of Reducing Antimicrobial Use and Applying a Cleaning and Disinfection Program in Veal Calf Farming: Experiences from an Intervention Study to Control Livestock-Associated MRSA.

With the ultimate aim of containing the emergence of resistant bacteria, a Dutch policy was set in place in 2010 promoting a reduction of antimicrobial use (AMU) in food-producing animals. In this context, a study evaluated strategies to curb livestock-associated methicillin resistant Staphylococcus aureus (LA-MRSA). Fifty-one veal calf farms were assigned to one of 3 study arms: RAB farms reducing antimicrobials by protocol; RAB-CD farms reducing antimicrobials by protocol and applying a cleaning and disinfection program; and Control farms without interventions. MRSA carriage was tested in week 0 and week 12 of 2 consecutive production cycles in farmers, family members and veal calves. Interventions were validated and a cyclic rise in MRSA-prevalence in animals was shown with a more moderate increase in RAB farms. Prevalence in humans declined parallel over time in the study arms but RAB farms were at the lowest MRSA levels from the beginning of the study. In RAB-CD farms, human and animal prevalence did not differ from Control farms and MRSA air loads were significantly higher than in the other study arms. Mimicking the national trend, an overall AMU decrease (daily dosages per animal per cycle (DDDA/C)) was observed over 4 pre-study and the 2 study cycles; this trend did not have a significant effect on a set of evaluated farm technical parameters. AMU was positively associated with MRSA across study arms (ORs per 10 DDDA/C increase = 1.26 for both humans (p = 0.07) and animals (p = 0.12 in first cycle)). These results suggest that AMU reduction might be a good strategy for curbing MRSA in veal calf farming, however the specific cleaning and disinfecting program in RAB-CD farms was not effective. The drop in MRSA prevalence in people during the study could be attributed to the observed long-term AMU decreasing trend.

Livestock-associated MRSA in household members of pig farmers: transmission and dynamics of carriage, a prospective cohort study.

This prospective cohort study describes carriage of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) in household members from 49 farrowing pig farms in the Netherlands (2010-2011). Of 171 household members, 4% were persistent MRSA nasal carriers, and the MRSA prevalence on any given sampling moment was 10% (range 7-11%). Working in the stables (of which 98% was MRSA-positive, prevalence ratio (PR) = 2.11 per 10 hours), working with sows (PR=1.97), and living with an MRSA-positive pig farmer (PR=4.63) were significant determinants for MRSA carriage. Significant protective factors were carriage of methicillin-susceptible Staphylococcus aureus (MSSA) (PR=0.50), and wearing a facemask when working in the stables (37% decreased prevalence). All MRSA strains during the study period were known livestock-associated types. The bacteriophage φ3 was not found in household members. Transmission from pigs and the environment appeared to be important determinants; human-to-human transmission could not sufficiently be differentiated. Wearing a facemask when working in the stables and carriage of MSSA are potential interventional targets.

Consumption of antimicrobials in pigs, veal calves, and broilers in the Netherlands: quantitative results of nationwide collection of data in 2011.

In 2011, Dutch animal production sectors started recording veterinary antimicrobial consumption. These data are used by the Netherlands Veterinary Medicines Authority to create transparency in and define benchmark indicators for veterinary consumption of antimicrobials. This paper presents the results of sector wide consumption of antimicrobials, in the form of prescriptions or deliveries, for all pig, veal calf, and broiler farms. Data were used to calculate animal defined daily dosages per year (ADDD/Y) per pig or veal calf farm. For broiler farms, number of animal treatment days per year was calculated. Furthermore, data were used to calculate the consumption of specific antimicrobial classes per administration route per pig or veal calf farm. The distribution of antimicrobial consumption per farm varied greatly within and between farm categories. All categories, except for rosé starter farms, showed a highly right skewed distribution with a long tail. Median ADDD/Y values varied from 1.2 ADDD/Y for rosé finisher farms to 83.2 ADDD/Y for rosé starter farms, with 28.6 ADDD/Y for white veal calf farms. Median consumption in pig farms was 9.3 ADDD/Y for production pig farms and 3.0 ADDD/Y for slaughter pig farms. Median consumption in broiler farms was 20.9 ATD/Y. Regarding specific antimicrobial classes, fluoroquinolones were mainly used on veal calf farms, but in low quantities: P75 range was 0 - 0.99 ADDD/Y, and 0 - 0.04 ADDD/Y in pig farms. The P75 range for 3(rd)/4(th)-generation cephalosporins was 0 - 0.07 ADDD/Y for veal calf farms, and 0 - 0.1 ADDD/Y for pig farms. The insights obtained from these results, and the full transparency obtained by monitoring antimicrobial consumption per farm, will help reduce antimicrobial consumption and endorse antimicrobial stewardship. The wide and skewed distribution in consumption has important practical and methodological implications for benchmarking, surveillance and future analysis of trends.

Livestock-associated MRSA prevalence in veal calf production is associated with farm hygiene, use of antimicrobials, and age of the calves.

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) is highly prevalent in pork and veal production chains. In this study, we used data from a cross-sectional survey on 2151 calves from 102 veal calf farms to identify potential risk factors, with the goal of reducing MRSA prevalence by developing intervention strategies. Overall, calves from rose veal farms had a lower risk of LA-MRSA carriage than calves from white veal farms. Data were analysed separately for white and rose veal calves, because management systems of the two production chains were largely different. Group treatment with antimicrobials appeared to be a risk factor for MRSA carriage in white veal calves in univariate analyses, but was not included in the final multiple regression model that included age of the calves and rodent control. Number of start treatment days was positively associated with LA-MRSA carriage in rose veal calves, and was the only risk factor selected for the final multiple regression model for this group. Interpretation of the results from this cross-sectional study is complicated by the strong correlation between antimicrobial use, LA-MRSA carriage and age of the calves. Other age-related factors may be more influential. However, taken together these findings emphasize the need for prudent use of antimicrobials, and point to improvement of farm hygiene as a control measure.

Dynamics of MRSA carriage in veal calves: a longitudinal field study.

Colonization of Methicillin-Resistant Staphylococcus aureus (MRSA) in food producing animals has public health implications, but intervention targets have not yet been identified. In this field study occurrence and dynamics of MRSA in veal calves were investigated longitudinally on three farms. Determinants generally associated with MRSA carriage, such as environmental exposure and antimicrobial use, were explored. In addition, the reliability and reproducibility of MRSA detection in nasal samples from veal calves were investigated as well as the additional value of rectal samples to establish MRSA status of an individual animal. On these three farms, MRSA prevalence and MRSA air loads in stables rapidly increased during the production cycle, especially after releasing calves from their individual houses, but not simultaneously with or directly after treatment with antimicrobials. These observations constitute the hypothesis that antimicrobial use may not necessarily be the only condition for MRSA transmission in veal calves, but indicate that other factors may contribute to transmission as well. MRSA in calves was present both nasally and rectally. The reproducibility and repeatability of the nasal samples were moderate. The results of this study give a better understanding of the dynamics of MRSA in a field situation.

Within-flock transmission of H7N1 highly pathogenic avian influenza virus in turkeys during the Italian epidemic in 1999-2000.

Highly pathogenic avian influenza (HPAI) viruses show different disease dynamics between virus strains and host species, and therefore epidemic field data are very valuable. This study used field data of the HPAI H7N1 epidemic which affected Northern Italy in 1999-2000. Field mortality data was back-calculated into a S-I-format to estimate the transmission rate parameter beta, indicating the number of birds infected per infectious bird per time unit. The bird-infectious period was assumed to be 2 days, and all birds were assumed to die after this infectious period. The estimated beta for HPAI H7N1 virus transmission in turkeys was 1.43 (95% CI: 1.17-1.74). Farm risk factors such as flock size and age of the turkeys did not influence the estimated transmission rate parameter.

Effect of H7N1 vaccination on highly pathogenic avian influenza H7N7 virus transmission in turkeys.

This study describes the results of a transmission experiment with highly pathogenic avian influenza (HPAI) H7N7 virus in 12-week-old turkeys. Cloacal and tracheal swabs as well as serum samples were taken to monitor the infection both in inoculated and in susceptible contact turkeys, which were all either unvaccinated, vaccinated once or vaccinated twice with H7N1. Swabs were tested by real-time RT-PCR and serum samples with hemagglutination inhibition test (HI). Unvaccinated contact birds had a mean infectious period of 6.2 days, and an estimated transmission rate parameter of 1.26 per infectious bird per day. However, no virus shedding was found in inoculated vaccinated turkeys and thus we concluded that vaccination with H7N1 protected against challenge with HPAI H7N7 virus.

Estimating the day of highly pathogenic avian influenza (H7N7) virus introduction into a poultry flock based on mortality data.

Despite continuing research efforts, knowledge of the transmission of the highly pathogenic avian influenza (HPAI) virus still has considerable gaps, which complicates epidemic control. The goal of this research was to develop a model to back-calculate the day HPAI virus is introduced into a flock, based on within-flock mortality data. The back-calculation method was based on a stochastic SEIR (susceptible (S) - latently infected (E) - infectious (I) - removed (= dead; R)) epidemic model. The latent and infectious period were assumed to be gamma distributed. Parameter values were based on experimental H7N7 within-flock transmission data. The model was used to estimate the day of virus introduction based on a defined within-flock mortality threshold (detection rule for determining AI). Our results indicate that approximately two weeks can elapse before a noticeable increase in mortality is observed after a single introduction into a flock. For example, it takes twelve (minimum 11 - maximum 15) days before AI is detected if the detection rule is fifty dead chickens on two consecutive days in a 10 000 chicken flock (current Dutch monitoring rule for notification). The results were robust for flock size and detection rule, but sensitive to the length of the latent and infectious periods. Furthermore, assuming multiple introductions on one day will result in a shorter estimated period between infection and detection. The implications of the model outcomes for detecting and tracing outbreaks of H7N7 HPAI virus are discussed.