Draft Genome Sequences of Three Porcine Streptococcus suis Isolates Which Differ in Their Susceptibility to Penicillin.

The draft genome sequences of three Streptococcus suis isolates, IMT40343, IMT40201, and IMT40738, are presented here. These isolates were obtained from bronchoalveolar lavage fluid of healthy and diseased weaners from different German piglet-producing farms and differed in their susceptibility to penicillin.

Plasmid-located dfrA14 gene in Pasteurella multocida isolates from three different pig-producing farms in Germany.

Pasteurella multocida is an important respiratory tract pathogen in intensive livestock farming, especially in pigs. Antimicrobial agents are frequently used to combat infections caused by this pathogen. In a study on antimicrobial resistance among respiratory tract pathogens of pigs from 30 German pig-producing farms, P. multocida isolates (n = 9) with high minimal inhibitory concentration (MIC) values of 16/304 mg/L (n = 2), 32/608 mg/L (n = 3) or ≥64/1216 mg/L (n = 4) for trimethoprim/sulfamethoxazole (1:19) and of ≥512 mg/L (n = 9) for trimethoprim (TMP) were detected in three of these farms. The genetic relatedness of the isolates was investigated via capsule-specific PCR and macrorestriction analyses with ApaI and SmaI. Pulsed-field gel electrophoresis revealed indistinguishable restriction patterns per farm, with slight differences between the three farms. All isolates represented capsular type A. Four representative isolates, that were subjected to whole genome sequencing, shared the multi-locus sequence type (ST) 3. Their plasmids were transformed into E. coli TOP10 with subsequent selection on TMP-containing agar plates. Antimicrobial susceptibility testing and plasmid analysis of the transformants confirmed that they were resistant to sulfonamides and trimethoprim and carried only a single small plasmid. This plasmid was completely sequenced and revealed a size of 6050 bp. Sequence analyses identified the presence of a resistance gene cluster comprising the genes sul2-ΔstrA-dfrA14-ΔstrA-ΔstrB. Further analysis identified a dfrA14 gene cassette being integrated into the strA reading frame. Neither the gene dfrA14 nor this gene cluster have been detected before in P. multocida.

Antimicrobial susceptibility and genetic relatedness of respiratory tract pathogens in weaner pigs over a 12-month period.

The collaboration project VASIB aims at reducing the antibiotic consumption in pig production by integrating information from consulting expertise in clinical inspection, hygiene, epidemiology, microbiology and pharmacology. In this VASIB subproject, we investigated the antimicrobial susceptibility and relatedness of porcine respiratory tract pathogens. Bordetella bronchiseptica (n = 47), Pasteurella multocida (n = 18) and Streptococcus suis (n = 58) were obtained from weaner pigs at two farms. Antimicrobial susceptibility testing was performed by broth microdilution according to CLSI standards. Resistance genes were detected via specific PCR assays. Macrorestriction analysis was conducted to determine the relatedness of the isolates and to identify clones. The B. bronchiseptica isolates showed indistinguishable (farm 1) or two closely related XbaI-patterns (farm 2). Different SmaI-PFGE patterns of P. multocida isolates were obtained at three different time points. In contrast, PFGE analysis of S. suis indicated more than one fragment pattern per pig and time point. Isolates exhibiting indistinguishable PFGE patterns were considered to represent the same clone. This study showed that only two closely related B. bronchiseptica clones were present in both farms, which had low MICs to all antimicrobials, except to ß-lactams. Different P. multocida clones were present at the three time points. They showed overall low MIC values, with two clones being resistant and one intermediate to tetracycline. S. suis clones were resistant to tetracycline (n = 19) and/or erythromycin/clindamycin (n = 16). They harboured the tetracycline resistance genes tet(O), tet(M) or tet(L) and/or the macrolide/lincosamide/streptogramin B resistance gene erm(B). Five penicillin-resistant S. suis clones were also detected.

[Investigations into the use of respiratory masks for reducing the MRSA-exposure of veterinarians visiting regularly pig herds--first experiences]. / Zum Einsatz von Staubmasken zur Senkung der MRSA-Exposition von Tieräirzten in der Bestandsbetreuung von Schweinebeständen- eine Pilotstudie.

The study presents first experiences on the controlled use of respiratory masks against Methicillin-resistant Staphylococcus aureus (MRSA) in a multi-person veterinary pig practice. Seven veterinarians entered the trial (five wearing masks, two wearing no masks) after the veterinarians had performed a decolonisation protocol. The pig herds were visited regularly by the veterinarians during the study period. The five "trial" veterinarians wore gloves and respiratory masks for at least 30 days and 30 farm visits. The two "control" veterinarians wore gloves only. Nasal swabs were collected at a seven day interval. Swabs and ten masks per "trial" veterinarian were bacteriologically tested for MRSA including MLST- and spa-typing. The study showed a high MRSA-exposure for the veterinarians, since 68% of the masks were tested positive for MRSA. However, four vets stayed MRSA-negative while using the masks. Only one of the"trial" veterinarians became positive after two weeks. After the masks were not worn any more, two veterinarians returned to colonisation soon again. The two "control" veterinarians turned positive after 26 and 54 days, respectively. The high finding-rates of MRSA in the masks proof an enormous risk of nasal colonisation during routine work.The results of our study do not proof the potential of respiratory masks to prevent nasal colonisation of veterinarians with MRSA. However, there are no hints, that the proper use of masks could be a risk factor for becoming colonised. Further details of the proper use of masks and the quantification of their protective potential need further studies on a larger scale.

[Intra-herd prevalence and colonisation dynamics of Methicillin-resistant Staphylococcus aureus (MRSA) in two pig breeding herds]. / Intra-herdenprävalenz und Kolonisa- tionsdynamik von Methicillin-resistenten Staphylococcus aureus in zwei Schweine- zuchtbeständen.

This study presents data on the intra-herd prevalence and colonisation dynamics of Methicillin-resistant (MRSA) and Methicillin-sensible (MSSA) Staphylococcus aureus in two independent pig herds located in the Northwest of Germany. Swabs taken from the nasal cavity of sows and piglets, from the udder of the sows and from the direct environment of the pigs were tested microbiologically for MRSA and MSSA. The piglets were identified individually, tested at birth and then swabbed again every 3 weeks from birth to sale at the end of nursery with 25 kg (farm A), and at slaughter respectively (farm B). The intra-herd prevalence differed between the two farms remarkably. In addition, both farms differed concerning the point in time of the first detection of MRSA in piglets: While MRSA was detected in piglets on farm A within three days after farrowing, positive results in piglets did not occur before three weeks of life on farm B. An MRSA-positive result of the sow increased the number of MRSA-positive piglets on days 1-3. Moreover the results of this study support the suggestion, that an early colonisation of the piglets with Methicillin-sensitive S. aureus (MSSA) has a protective effect. Over time, the pigs changed their pigs revealed that the proper cleaning and disinfection on one of the study farms had a reducing effect on the MRSA occurrence in the farrowing pens and in the flat deck.

Nasal colonization of humans with methicillin-resistant Staphylococcus aureus (MRSA) CC398 with and without exposure to pigs.

BACKGROUND: Studies in several European countries and in North America revealed a frequent nasal colonization of livestock with MRSA CC398 and also in humans with direct professional exposure to colonized animals. The study presented here addresses the question of further transmission to non exposed humans. METHODS: After selecting 47 farms with colonized pigs in different regions of Germany we sampled the nares of 113 humans working daily with pigs and of their 116 non exposed family members. The same was performed in 18 veterinarians attending pig farms and in 44 of their non exposed family members. For investigating transmission beyond families we samples the nares of 462 pupils attending a secondary school in a high density pig farming area. MRSA were detected by direct culture on selective agar. The isolates were typed by means of spa-sequence typing and classification of SCCmec elements. For attribution of spa sequence types to clonal lineages as defined by multi locus sequence typing we used the BURP algorithm. Antibiotic susceptibility testing was performed by microbroth dilution assay. RESULTS: At the farms investigated 86% of humans exposed and only 4.3% of their family members were found to carry MRSA exhibiting spa-types corresponding to clonal complex CC398. Nasal colonization was also found in 45% of veterinarians caring for pig farms and in 9% of their non exposed family members. Multivariate analysis revealed that antibiotic usage prior to sampling beard no risk with respect to colonization. From 462 pupils only 3 were found colonized, all 3 were living on pig farms. CONCLUSION: These results indicate that so far the dissemination of MRSA CC398 to non exposed humans is infrequent and probably does not reach beyond familial communities.