Search

Globally, antimicrobial resistance is one of the most important public health challenges in which the clinical microbiology laboratory plays a critical role by providing guidance for antimicrobial treatment. Despite the recognition of its importance, there is still a real need for the standardized training of clinical microbiologists and harmonization of diagnostic procedures. This is particularly true for veterinary clinical microbiology, where additional challenges exist when microbiologists are trying to fulfill a professional role very similar to that of their colleagues working in human microbiology laboratories. The specific points that need addressing to improve the outputs of veterinary microbiology laboratories discussed here include (i) harmonization of methodologies used by veterinary laboratories for antimicrobial susceptibility testing (AST); (ii) specific guidelines for interpretation and reporting of AST results for animal pathogens; (iii) guidelines for detection of antimicrobial resistance mechanisms in animal isolates; (iv) standardization of diagnostic procedures for animal clinical specimens; and (v) the need to train more veterinary clinical microbiology specialists. However, there is now a plan to address these issues, led by the European Network for Optimization of Veterinary Antimicrobial Treatment (ENOVAT), which is bringing together experts in veterinary microbiology, pharmacology, epidemiology, and antimicrobial stewardship from Europe and wider afield. ENOVAT is aiming to work with project partners toward standardization and harmonization of laboratory methodologies and optimization of veterinary antimicrobial treatment. Ultimately, the project may provide a mechanism for standardization and harmonization of veterinary clinical microbiology methodologies that could then be used as a template for implementation at a wider international level.

Background In the Netherlands there is a lack of data regarding resistance of Staphylococcus pseudintermedius to the systemic antimicrobial drugs used for the treatment of superficial pyoderma. Objectives To assess antimicrobial resistance, with emphasis on resistance to clindamycin and meticillin, in clinical isolates of S. pseudintermedius isolated from dogs with superficial pyoderma. Results were compared between dogs with and without a history of systemic antimicrobial therapy during the previous year. Animals A retrospective study of 237 referral cases presented to an academic teaching hospital between 2014 and 2019, with the clinical and microbiological diagnosis of superficial pyoderma. Methods and materials All clinical isolates were identified primarily by MALDI‐TOF mass spectrometry. Antimicrobial susceptibility was tested either by an agar diffusion method (2014–2016) or by broth microdilution. Antimicrobial history in the preceding year was obtained from medical records. Results Meticillin‐resistant S. pseudintermedius (MRSP) was isolated from 8% of superficial pyoderma cases. Within the meticillin‐susceptible S. pseudintermedius (MSSP) population, clindamycin resistance was significantly more common in isolates derived from dogs with histories of antimicrobial treatment (37.7%) compared to dogs with no histories of exposure (21.7%; P = 0.03). Conclusions Given the high prevalence of clindamycin resistance in MSSP isolated from dogs with prior antimicrobial exposure, it is recommended that bacterial culture and susceptibility testing be pursued before prescribing systemic antimicrobials. Clindamycin should be regarded as the preferred treatment option if susceptibility is confirmed, due to its narrow spectrum and reduced selective pressure for MRSP., Background – In the Netherlands there is a lack of data regarding resistance of Staphylococcus pseudintermedius to the systemic antimicrobial drugs used for the treatment of superficial pyoderma. Objectives – To assess antimicrobial resistance, with emphasis on resistance to clindamycin and meticillin, in clinical isolates of S. pseudintermedius isolated from dogs with superficial pyoderma. Results were compared between dogs with and without a history of systemic antimicrobial therapy during the previous year. Conclusions – Given the high prevalence of clindamycin resistance in MSSP isolated from dogs with prior antimicrobial exposure, it is recommended that bacterial culture and susceptibility testing be pursued before prescribing systemic antimicrobials. Clindamycin should be regarded as the preferred treatment option if susceptibility is confirmed, due to its narrow spectrum and reduced selective pressure for MRSP.

Since the first report on methicillin resistant Staphylococcus aureus (MRSA) CC398 in pigs, several countries have determined the prevalence of MRSA-positive pig herds using different sampling and laboratory techniques. The objective of the study was to compare three sampling methods for MRSA-classification of herds. Therefore, nasal swabs of pigs and environmental wipes were collected from 147 herds with breeding pigs. Per herd, laboratory examination was done on 10 pools of 6 nasal swabs (NASAL), 5 single environmental wipes (ENVSINGLE) and one pool of 5 environmental wipes (ENVPOOL). Large differences in apparent prevalence of MRSA-positive herds between methods were found: 19.1% for ENVPOOL, 53.1% for ENVSINGLE, and 70.8% for NASAL. Pairwise comparisons of methods resulted in relative sensitivities of 26.9% (ENVPOOL vs. NASAL), 34.6% (ENVPOOL vs. ENVSINGLE), and 72.1% (ENVSINGLE vs. NASAL) with relative specificities of respectively 100%, 98.6% and 93.0%. Cohen's kappa was respectively 0.18, 0.32 and 0.55, thus varying between very poor and moderate agreement. Examination of environmental wipes is an easy and non-invasive method to classify herds for MRSA. The number of environmental wipes needed depends on e.g. required detection limits and within-herd prevalence. In low prevalent herds (e.g. herds with

The prevalence of methicillin resistant Staphylococcus aureus (MRSA) in pigs at abattoirs is higher than in pigs sampled on farms. This study investigated whether MRSA negative pigs can become MRSA positive during transportation from the farm to the abattoir after exposure to other pigs and environmental sources of MRSA. Nasal swabs were collected from four batches of pigs during loading at the farm, on arrival at the abattoir and after stunning. Environmental wipes were taken from lorries after transporting pigs and from lairages after holding pigs. All pigs (n = 117) tested MRSA negative before transportation. On arrival at the abattoir, 12/117 (10.3%) pigs in two batches tested MRSA positive. In lorries that tested positive after transportation, the prevalence of MRSA positive pigs was 21.1%, whereas no MRSA was detected in pigs that had been transported in lorries that tested negative after transportation. At stunning, all batches and 70/117 (59.8%) pigs tested MRSA positive. Pigs can become MRSA positive in the short period of time during transportation from the farm to stunning at the abattoir.

SUMMARYLivestock-associated MRSA has been found in various animals, livestock farmers and retail meat. This study aimed to determine the prevalence and determinants of nasal MRSA carriage in pig slaughterhouse workers. Three large pig slaughterhouses in The Netherlands were studied in 2008 using human and environmental samples. The overall prevalence of nasal MRSA carriage in employees of pig slaughterhouses was 5·6% (14/249) (95% CI 3·4–9·2) and working with live pigs was the single most important factor for being MRSA positive (OR 38·2,Pr=0·75). In conclusion, a high prevalence of nasal MRSA carriage was found in pig-slaughterhouse workers, and working with live pigs is the most important risk factor. Exact transmission routes from animals to humans remain to be elucidated in order to enable application of targeted preventive measures.

In The Netherlands, MRSA ST398 has emerged in hospitals and human carriers have been associated with exposure to pigs and cattle. High prevalences of MRSA ST398 in pigs and pig farmers have been determined and the transmission routes of MRSA on pig farms need to be elucidated. In the south of the Netherlands, in recent years, the black rat (Rattus rattus) has emerged as a prominent rodent on livestock farms. From March till May 2008, a survey on MRSA in rats living on livestock farms in the south of The Netherlands and the north of Belgium was conducted. In total, 40 black rats (R. rattus) and 3 brown rats (Rattus norvegicus) were collected on 12 farms including five pig farms, five poultry farms, one mixed pig and veal farm and one goat farm. MRSA ST398 was detected in black rats captured at two of the five pig farms as well as in a black rat living on the mixed pig and veal farm. From one black rat captured at another pig farm MRSA ST 97 was isolated. Considering the behaviour of rats on livestock farms, it is concluded that rats might play a role in the spread and persistence of MRSA on pig farms.

International surveillance of antimicrobial use in food animal production shows that methicillin-resistant Staphylococcus aureus (MRSA), traditionally a human pathogen associated with hospitals, has emerged in the community and animals. Since 1961, MRSA has been causing human infections in hospitals worldwide and a vast majority of them were caused by five major epidemic clones. After 1990, other clones have emerged in the community, leading to infections in relatively young and healthy individuals. The origin of these clones is largely unknown, and extensive diversity among isolates exists. Companion animals have been indicated as a reservoir. However, most studies suggest that they are initially infected by humans and subsequently animals re-colonize humans. More recently, a new zoonotic reservoir in food production animals was found. This involves a specific clone, MRSA ST398, which spreads extensively in animals and is also found in retail meat. It poses a potential threat to public health, as people in contact with food production animals are at much higher risk of colonization. The most probable transmission route seems to be by (in)direct contact, as dust in stables was found positive for MRSA ST398. The role of MRSA ST398 as a food pathogen needs more research. To prevent colonization in humans, it is important to investigate transmission routes and transmission dynamics between animals, between animals and humans and between humans. Collaboration of human and veterinary epidemiologists and microbiologists is needed to identify the implications of this strain for public health and to develop cost-effective control strategies.

Results: These 16 herds were sampled 177 times, varying from 4 to 13 times per herd. 72.9% of all samplings had at least one positive sample. Per herd this ranged from 38% to 100%. A total of 882 dust swabs were collected, of which 32.8% were positive, varying from 12% to 58% per herd. 284 isolates were spa-typed, yielding 17 different spatypes, mostly t011 and t108, with a minimum of 1 and a maximum of 6 spa-types per herd for the entire period and with a maximum of 3 per sampling. MLVA-typing did not lead to much more information, only in one herd there were two different types of spa-type t011: t011-398 (common) and t011-555 (only once). In 12 herds there was a spa-type which was found from beginning to the end of the test period. In 4 herds the same spa-type was found, and still prominent, as was found in the initial sampling in 2008. In many herds there were incidental findings of rare spa-types. Detailed results can be found in table 1.

Extended-spectrum-β-lactamase (ESBL)/AmpC producing Enterobacteriaceae have been reported worldwide amongst isolates obtained from humans, food-producing animals, companion animals, and environmental sources. However, data on prevalence of fecal carriage of ESBL/AmpC producing Enterobacteriaceae in healthy companion animals is limited. This pilot study describes the prevalence of ESBL/AmpC encoding genes in healthy cats and dogs, and cats and dogs with diarrhea. Twenty fecal samples of each group were cultured on MacConkey agar supplemented with 1 mg/L cefotaxime and in LB-enrichment broth supplemented with 1 mg/L cefotaxime, which was subsequently inoculated on MacConkey agar supplemented with 1 mg/L cefotaxime. ESBL/AmpC genes were identified using the Check-Points CT103 micro array kit and subsequently by sequencing analysis. Chromosomal ampC promoter mutations were detected by PCR and sequencing analysis. From the healthy and diarrheic dogs, respectively 45 and 55% were positive for Escherichia coli with reduced susceptibility for cefotaxime. From the healthy and diarrheic cats, the estimated prevalence was respectively 0 and 25%. One diarrheic cat was positive for both reduced susceptible E. coli and Proteus mirabilis. The ESBL/AmpC genes found in this study were mainly bla CTX-M-1, but also bla CTX-M-14, bla CTX-M-15, bla TEM-52-StPaul, bla SHV-12, and bla CMY-2 were detected. This pilot study showed that the prevalence of ESBL/AmpC producing Enterobacteriaceae in healthy and diarrheic dogs, and diarrheic cats was relatively high. Furthermore, the genes found were similar to those found in isolates of both human and food-producing animal origin. However, since the size of this study was relatively small, extrapolation of the data to the general population of cats and dogs should be done with great care.

Meticillin-resistant Staphylococcus aureus (MRSA) clonal complex (CC) 398 is a genetic lineage associated with livestock, especially pigs. The authors investigated the role of pig trade in the transmission of MRSA CC398 between farms using pulsed-field gel electrophoresis (PFGE), a highly discriminatory method for strain typing. PFGE analysis of 58 MRSA isolates from a retrospective study in the Netherlands and a prospective study in Denmark provided molecular evidence that the strains present in five of the eight recipient farms were indistinguishable from those occurring in the corresponding supplying farm. The molecular typing data confirm the findings of a previous risk-analysis study indicating that trading of colonised pigs is a vehicle for transmission of MRSA CC398.

A PCR targeting sau1-hsdS1 was developed for rapid detection of Staphylococcus aureus clonal complex 398 (CC398). High sensitivity (100%) and specificity (100%) were shown by evaluating the test on a large strain collection ( n = 1,307). We recommend this test for accurate, rapid, and inexpensive diagnosis of methicillin-resistant S. aureus (MRSA) CC398 in hospitals and on farms.

Methicillin-resistant Staphylococcus aureus clonal complex (CC) 398 has emerged from pigs to cause human infections in Europe and North America. We used a new 62-strain S. aureus microarray (SAM-62) to compare genomes of isolates from three geographical areas (Belgium, Denmark, and Netherlands) to understand how CC398 colonizes different mammalian hosts. The core genomes of 44 pig isolates and 32 isolates from humans did not vary. However, mobile genetic element (MGE) distribution was variable including SCCmec. ϕ3 bacteriophage and human specificity genes (chp, sak, scn) were found in invasive human but not pig isolates. SaPI5 and putative ruminant specificity gene variants (vwb and scn) were common but not pig specific. Virulence and resistance gene carriage was host associated but country specific. We conclude MGE exchange is frequent in CC398 and greatest among populations in close contact. This feature may help determine epidemiological associations among isolates of the same lineage., Journal Article, SCOPUS: ar.j, info:eu-repo/semantics/published

In 2005, methicillin-resistant Staphylococcus aureus was found in pig herds and in humans in contact with pigs. To determine the prevalence of, this now-called livestock-associated (LA) MRSA among pig herds in The Netherlands and to identify and quantify risk factors, an observational study of 202 pig herds was performed between 2007 and 2008. Five environmental wipes and 60 nasal swabs from each herd were collected, and microbiological analysis was performed on single environmental samples and pooled nasal samples. A herd was considered MRSA-positive if ≥1 sample tested positive. The prevalence of MRSA-positive herds was 67% in breeding herds and 71% in finishing herds. Multivariable logistic regression analysis was then performed on data from 171 breeding herds. The number of MRSA-positive herds increased from ∼30% at the start to ∼75% at the end of the study, most likely due to transmission between herds. The prevalence of MRSA increased with herd size, as ∼40% of smaller herds (250 sows) were MRSA-positive compared to80% of larger herds (500 sows). Other risk factors (e.g. antimicrobial use, purchase of gilts and hygiene measures) were not significantly associated with MRSA, though associated with herd size. Herd size appeared to be a compilation of several factors, which made larger herds more often MRSA positive.

SUMMARYWe compared the prevalence of human and animal methicillin-resistantStaphylococcus aureus(MRSA) at pig farms in The Netherlands, and related this to individual and farm-level characteristics. More than half of the farms investigated (28/50) had MRSA in pigs or stable dust and about one third (15/50) of person(s) were identified as MRSA carriers. Human carriage was found only on farms with MRSA-positive pigs or dust. MRSA strains in human samples were the samespa-type as found in pigs and all were not typable by pulsed-field gel electrophoresis (NT-MRSA). Multivariate analyses showed that risk factors for human MRSA carriage were: working in pig stables (OR 40, 95% CI 8–209) and the presence of sows and finishing pigs (OR 9, 95% CI 3–30). Veterinary sample collectors sampling the pigs showed transient MRSA carriage only during the day of the farm visit. Working in pig stables with MRSA-positive pigs poses a high risk for acquiring MRSA, increasingly so when contact with live pigs is more intensive or long lasting.