Haitske Graveland

Methicillin Resistant Staphylococcus aureus ST398 in Veal Calf Farming : Human MRSA Carriage Related with Animal Antimicrobial Usage and Farm Hygiene

Introduction Recently a specific MRSA sequence type, ST398, emerged in food production animals and farmers. Risk factors for carrying MRSA ST398 in both animals and humans have not been fully evaluated. In this cross-sectional study, we investigated factors associated with MRSA colonization in veal calves and humans working and living on these farms. Methods A sample of 102 veal calf farms were randomly selected and visited from March 2007–February 2008. Participating farmers were asked to fill in a questionnaire (n = 390) to identify potential risk factors. A nasal swab was taken from each participant. Furthermore, nasal swabs were taken from calves (n = 2151). Swabs were analysed for MRSA by selective enrichment and suspected colonies were confirmed as MRSA by using slide coagulase test and PCR for presence of the mecA-gene. Spa types were identified and a random selection of each spa type was tested with ST398 specific PCR. The Sequence Type of non ST398 strains was determined. Data were analyzed using logistic regression analysis. Results Human MRSA carriage was strongly associated with intensity of animal contact and with the number of MRSA positive animals on the farm. Calves were more often carrier when treated with antibiotics, while farm hygiene was associated with a lower prevalence of MRSA. Conclusion This is the first study showing direct associations between animal and human carriage of ST398. The direct associations between animal and human MRSA carriage and the association between MRSA and antimicrobial use in calves implicate prudent use of antibiotics in farm animals.

Livestock-associated methicillin-resistant Staphylococcus aureus in animals and humans

Since 2004 MRSA emerged in animals, particularly in pigs and veal calves. This new MRSA variant was since its first appearance referred to as Livestock Associated-MRSA (LA-MRSA). In Europe and Northern America, LA-MRSA belongs predominantly to clonal complex (CC) 398 whereas in Asia ST9 seems to be dominant in pigs. Persons in direct contact with LA-MRSA-positive animals have an increased risk of becoming MRSA positive. The risk of carriage is mainly related with the intensity of animal contact and with MRSA prevalence among animals on the farm. In contrast with its success in animals, it seemed that MRSA CC398 is a poor persistent colonizer in humans. MRSA ST398 can, however, cause serious (invasive) infections and outbreaks, although, only incidentally reported so far. Farm hygiene and antimicrobial use contributed to MRSA occurrence in animals. Therefore these two determinants should in principle be incorporated into MRSA-control programmes in animal production. Like any other microorganism, LA-MRSA is expected to be able to adapt to new hosts and may change over time in the potential to colonize and to produce toxins. Also, the current circulating clone CC398 may be replaced by another clone in Western countries or emerge in countries where this clone is currently low-prevalent. Ongoing MRSA surveillance in humans and animals is needed to detect changes in epidemiology and to implement effective control measures.

Persistence of Livestock Associated MRSA CC398 in Humans Is Dependent on Intensity of Animal Contact

Introduction The presence of Livestock Associated MRSA (LA-MRSA) in humans is associated with intensity of animal contact. It is unknown whether the presence of LA-MRSA is a result of carriage or retention of MRSA-contaminated dust. We conducted a longitudinal study among 155 veal farmers in which repeated nasal and throat swabs were taken for MRSA detection. Periods with and without animal exposure were covered. Methods Randomly, 51 veal calf farms were visited from June - December 2008. Participants were asked to fill in questionnaires (n = 155) to identify potential risk factors for MRSA colonisation. Nasal and throat swabs were repeatedly taken from each participant for approximately 2 months. Swabs were analysed for MRSA and MSSA by selective bacteriological culturing. Spa-types of the isolates were identified and a ST398 specific PCR was performed. Data were analyzed using generalized estimation equations (GEE) to allow for correlated observations within individuals. Results Mean MRSA prevalence was 38% in farmers and 16% in family members. Presence of MRSA in farmers was strongly related to duration of animal contact and was strongly reduced in periods with absence of animal contact (−58%). Family members, especially children, were more often carriers when the farmer was a carrier (OR = 2, P<0.05). Only 7% (n = 11) of the participants appeared to be persistent carriers. A large heterogeneity in spa-types was detected, however 92.7% belonged to LA-MRSA CC398. A surprisingly high fraction of the spa-types (7.3%) did not belong to CC398. Conclusion The presence of LA-MRSA in farmers is strongly animal-exposure related. The rapidly decreasing MRSA prevalence during absence of animal contact suggests that LA-MRSA is a poor persistent colonizer in most humans. These results are of relevance for MRSA control strategies.

Persistence of Livestock-Associated Methicillin-Resistant Staphylococcus aureus in Field Workers after Short-Term Occupational Exposure to Pigs and Veal Calves

ABSTRACT The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) carriage in pig and veal calf farmers in the Netherlands is estimated at 25 to 35%. However, no information is available about MRSA carriage in humans after short-term occupational exposure to pigs or veal calves. This study examines the prevalence and duration of MRSA acquisition after short-term intensive exposure to pigs or veal calves for persons not exposed to livestock on a daily basis. The study was performed with field workers who took samples from the animals or the animal houses in studies on MRSA prevalence in pig and veal farms. They were tested for MRSA by taking nasal samples before, directly after, and 24 h after they visited the farms. There were 199 sampling moments from visits to 118 MRSA-positive farms. Thirty-four of these visits (17%) resulted in the acquisition of MRSA. Thirty-one persons (94%) appeared negative again after 24 h. There were 62 visits to 34 MRSA-negative farms; none of the field workers acquired MRSA during these visits. Except for that from one person, all spa types found in the field workers were identical to those found in the animals or in the dust in animal houses and belonged to the livestock-associated clone. In conclusion, MRSA is frequently present after short-term occupational exposure, but in most cases the strain is lost again after 24 h.

Evaluation of isolation procedures and chromogenic agar media for the detection of MRSA in nasal swabs from pigs and veal calves.

Since the emergence of MRSA in livestock, screening of animals for the detection of MRSA is widely practised. Different procedures are published for animal samples but a systematic comparison of methods has not been performed. The objective of this study was to compare three available commonly used procedures and three chromogenic agars for detecting MRSA in nasal swabs from pigs (n=70) and veal calves (n=100). Procedures 1 and 2 used a pre-enrichment comprising Mueller Hinton broth with 6.5% NaCl followed by selective enrichment with 4 microg/ml oxacillin+75 microg/ml aztreonam (procedure 1) and 5 microg/ml ceftizoxime+75 microg/ml aztreonam (procedure 2) respectively. Procedure 3 used a selective enrichment broth only, containing 4% NaCl, 5 microg/ml ceftizoxime+50 microg/ml aztreonam. After selective enrichment, media were streaked on to three different chromogenic agars. Significantly more MRSA were found for pig as well as for veal calf samples with procedures 1 and 2. No significant differences were found between procedures 1 and 2. For nasal swabs from pigs significantly more MRSA-positive samples were found when MRSA Screen (Oxoid) or MRSASelect (Bio-Rad) agars were used compared to MSRA ID (bioMérieux). For calf samples no significant differences between the different agars were found. In conclusion, the results of this study show that procedures 1 and 2, both using additional high salt pre-enrichment are superior and should be recommended for MRSA detection in nasal swabs from pigs and veal calves. The preferred choice of chromogenic agar depends on the sample matrix.

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.

Dynamics of methicillin-resistant Staphylococcus aureus and methicillin-susceptible Staphylococcus aureus carriage in pig farmers: a prospective cohort study

Our purpose was to determine the dynamics of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) carriage and its determinants in persons working at pig farms, in order to identify targets for interventions. This prospective cohort study surveyed 49 pig farms in the Netherlands on six sampling dates in 1 year (2010-11). Nasal and oropharyngeal swabs were collected, as well as environmental surface samples from stables and house. Of 110 pig farmers, 38% were persistent MRSA nasal carriers. The average cross-sectional MRSA prevalence was 63%. Methicillin-susceptible S. aureus (MSSA) nasal carriage was associated with fewer MRSA acquisitions (prevalence rate (PR) = 0.47, p 0.02). In multivariate analysis, an age of 40-49 years (PR = 2.13, p 0.01), a working week of ≥40 h (PR=1.89, p 0.01), giving birth assistance to sows (PR=2.26, p 0.03), removing manure of finisher pigs (PR=0.48, p 0.02), and wearing a facemask (PR = 0.13, p 0.02) were significantly related with persistent MRSA nasal carriage. A higher MRSA exposure in stables was associated with MRSA in pig farmers (p <0.0001). This study describes a very high prevalence of LA-MRSA carriage in pig farmers, reflecting extensive exposure during work. We identified the possible protective effects of MSSA carriage and of continuously wearing a facemask during work.

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.

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 β-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.

Air pollution and exhaled nitric oxide in Dutch schoolchildren

Background Short-term changes in air pollution exposure in children may be associated with transient increases in exhaled nitric oxide (NO), a marker of airway inflammation. Also, children living in areas with high air pollution levels and/or high traffic densities appear to have chronically increased levels of exhaled NO. No studies have simultaneously addressed the long-term and short-term associations between traffic-related air pollution and exhaled NO. Objectives To investigate associations between exhaled NO in school children and both short-term changes in outdoor PM10 and long-term traffic exposures. Methods Offline exhaled NO measurements were conducted in 812 children from nine Dutch schools within 400 m of motorways. Daily levels of particulate matter with a 50% cut-off aerodynamic diameter of 10 μm (PM10) were obtained from background monitoring stations. Long-term exposure to traffic-related air pollution was assessed using specific traffic-related characteristics such as total, car and truck motorway traffic and the distances of the childrens homes and schools from the motorway. Results A positive association was found between ambient PM10 concentrations on the day of exhaled NO measurement and exhaled NO (adjusted geometric means ratio (95% CI) 2.24 (1.37 to 3.65)) over the range of daily PM10 concentrations of 44 μg/m3), which was largely attributable to a pollution peak associated with high particulate matter emissions from traditional Easter fires. There were suggestive associations between exhaled NO and traffic counts only in children with asthma, which were not statistically significance. Conclusions Short-term changes in ambient PM10 largely attributable to biomass burning are associated with increased levels of exhaled NO.