Comparison of Antimicrobial Resistance Profiles in Salmonella spp. from Swine Upon Arrival and Postslaughter at the Abattoir.

Antimicrobial resistance (AMR) developed by Salmonella within animals used for food products is a major global issue. Monitoring AMR in animals destined for slaughter is, therefore, critical. Abattoirs may serve as potential candidate checkpoints for monitoring resistance patterns on farms. A complicating factor, however, is the impact of lairage on Salmonella detected in pigs at slaughter. This study sought to compare AMR patterns in Salmonella spp. in swine collected upon arrival (fecal samples) at the abattoir with those at postslaughter (cecal samples) and evaluate the feasibility of using slaughterhouse samples for surveillance of prevailing AMR Salmonella on farms. Eighty-four Salmonella isolates were recovered from a large, midwestern U.S. abattoir between September and November 2013. Isolates were tested for phenotypic AMR to 12 antimicrobials using the broth microdilution assay. Whole-genome sequencing identified the AMR genes harbored by the strains. Significant differences were observed in the isolate phenotypes and genotypes; however, no significant difference was observed in genotypic resistance patterns. Hence, the AMR profiles of Salmonella spp. postslaughter cannot be predicted from preslaughter samples. Further research considering the genetic diversity of isolates and statistical power of the genotypic analysis is warranted to improve the performance of WGS-inferred antimicrobial susceptibility.

Genotypes and Antimicrobial Susceptibility Profiles of Hemolytic Escherichia coli from Diarrheic Piglets.

Hemolytic Escherichia coli are important pathogens in neonatal and weaned pigs. In this study, we analyzed 95 hemolytic E. coli isolated from intestinal contents or fecal samples of diarrheic piglets in 15 states of the United States between November 2013 and December 2014. Phenotypic antimicrobial susceptibility was determined through Sensititre BOFO6F plates for all the strains. They were all resistant to clindamycin, penicillin, tiamulin, tilmicosin, and highly resistant to oxytetracycline (91.6%), chlortetracycline (78.9%), ampicillin (75.8%), and sulfadimethoxine (68.4%). 86.2% of them were multidrug resistant. Whole genome sequencing (WGS) showed that 55 strains were enterotoxigenic E. coli (ETEC) and 40 strains were non-ETEC, and the strains belonged to 22 known and 2 novel sequence types (STs). ST100 and ST10 were the main and predominant STs in ETEC strains, whereas the non-ETEC strains were diverse with ST23 and ST761 as the main STs. Antibiotic resistance gene/mutation profiling of the genomes confirmed the results of antimicrobial susceptibility test. Notably, significant differences were found in the susceptibility to enrofloxacin between ETEC and non-ETEC (58.2% vs. 5.0%) and gentamicin (32.7% vs. 7.5%). ampH, ampC2, and ampC1 were the most common beta-lactamase genes in all E. coli strains, and extended-spectrum beta-lactamase (ESBL) genes were rare in these isolates. This study provides new insights into antibiotic resistance and genotypes of intestinal pathogenic E. coli associated with swine disease in the United States, and support the utility of WGS in accurate prediction of resistance to most antibiotics.

Data evidencing slow anaerobic digestion in emergency treatment and disposal of infectious animal carcasses.

Burial of infectious and potentially infectious livestock and poultry animals is the most common response to an emergency situation. The data set summarizes 22-week-long experiment that simulates the environment found within conventional burial trenches for emergency disposal of animal carcasses, worldwide, sometimes with a topical application of quicklime as it is required in the Republic of Korea. This data set shows the rarely presented evidence of the extremely slow decay of animal carcasses. Besides visual evidence of no visible breakdown of carcass material, i.e., carcass (or carcass quarters and coarse cuts) still resembled the initial material at the end of the study, we present data characterizing the process. Specifically, temporal variations of digestate quality (pH, ammonia, volatile fatty acids), biogas production, and the persistence of odorous volatile organic compounds are summarized. The data provide important evidence of undesirable, slow progression of the digestion process. The evidence of failure to achieve practical endpoints with the anaerobic digestion provides the impetus for seeking alternative, improved methods of disposal that will be feasible in emergency context, such as aerated burial concept (Koziel et al., 2018 [1]).

Single Nucleotide Polymorphism Analysis Indicates Genetic Distinction and Reduced Diversity of Swine-Associated Methicillin Resistant Staphylococcus aureus (MRSA) ST5 Isolates Compared to Clinical MRSA ST5 Isolates.

Livestock associated methicillin resistant S. aureus (LA-MRSA) are lineages adapted to livestock species. LA-MRSA can be transmitted to humans and public health concerns exist because livestock may be the largest MRSA reservoir outside of hospital settings. Although the predominant European (ST398) and Asian (ST9) lineages of LA-MRSA are considered livestock adapted, North American swine also harbor ST5, a globally disseminated and highly pathogenic lineage. This study applied whole genome sequencing and single nucleotide polymorphism (SNP) typing to compare the population structure and genetic relatedness between swine associated and human clinical MRSA ST5 isolates. The established high-resolution phylogenomic framework revealed that LA-MRSA and human clinical MRSA ST5 are genetically distinct. LA-MRSA isolates were found to be clonal within farms, while greater genome diversity was observed among sampled clinical MRSA ST5. Analysis of the accessory genome demonstrated that LA-MRSA ST5 isolates and clinical MRSA ST5 isolates harbor different AMR genes and virulence factors, consistent with the SNP analysis. Collectively, our data indicate LA-MRSA and clinical MRSA ST5 isolates are distinct and the swine reservoir is likely of minimal significance as a source of clinical MRSA ST5 infections.

Antimicrobial Resistance Distribution Differs Among Methicillin Resistant Staphylococcus aureus Sequence Type (ST) 5 Isolates From Health Care and Agricultural Sources.

Antimicrobial resistance (AMR) is an expanding public health concern and methicillin resistant Staphylococcus aureus (MRSA) is a notable example. Since the discovery of livestock associated MRSA (LA-MRSA), public health concerns have arisen surrounding the potential of LA-MRSA isolates to serve as a reservoir for AMR determinants. In this study, we compare swine associated LA-MRSA ST5 and human clinical MRSA ST5 isolates for phenotypic antimicrobial susceptibilities determined via broth microdilution and genotypic determinants of AMR using whole genome sequencing and comparative genomic analysis to identify AMR elements. Swine associated LA-MRSA ST5 isolates exhibited phenotypic resistance to fewer antibiotics than clinical MRSA ST5 isolates from humans with no swine contact. Distinct genomic AMR elements were harbored by each subgroup, with little overlap in shared AMR genes between swine associated LA-MRSA ST5 and clinical MRSA ST5 isolates. Our results demonstrate that phenotypic antimicrobial susceptibilities and genotypic determinants of AMR among swine associated LA-MRSA ST5 and clinical MRSA ST5 isolates are separate and distinct.

Lab-scale evaluation of aerated burial concept for treatment and emergency disposal of infectious animal carcasses.

Nearly 55,000 outbreaks of animal disease were reported to the World Animal Health Information Database between 2005 and 2016. To suppress the spread of disease, large numbers of animal mortalities often must be disposed of quickly and are frequently buried on the farm where they were raised. While this method of emergency disposal is fast and relatively inexpensive, it also can have undesirable and lasting impacts (slow decay, concerns about groundwater contamination, pathogens re-emergence, and odor). Following the 2010 foot-and-mouth disease outbreak, the Republic of Korea's National Institute of Animal Science funded research on selected burial alternatives or modifications believed to have potential to reduce undesirable impacts of burial. One such modification involves the injection of air into the liquid degradation products from the 60-70% water from decomposing carcasses in lined burial trenches. Prior to prototype development in the field, a laboratory-scale study of aerated decomposition (AeD) of poultry carcasses was conducted to quantify improvements in time of carcass decomposition, reduction of potential groundwater pollutants in the liquid products of decomposition (since trench liners may ultimately leak), and reduction of odorous VOCs emitted during decomposition. Headspace gases also were monitored to determine the potential for using gaseous biomarkers in the aerated burial trench exhaust stream to monitor completion of the decomposition. Results of the lab-scale experiments show that the mass of chicken carcasses was reduced by 95.0 ±â€¯0.9% within 3 months at mesophilic temperatures (vs. negligible reduction via mesophilic anaerobic digestion typical of trench burial) with concomitant reduction of biochemical oxygen demand (BOD; 99%), volatile suspended solids (VSS; 99%), total suspended solids (TSS; 99%), and total ammonia nitrogen (TAN; 98%) in the liquid digestate. At week #7 BOD and TSS in digestate met the U.S. EPA standards for treated wastewater discharge to surface water. Salmonella and Staphylococcus were inactivated by the AeD process after week #1 and #3, respectively. Five gaseous biomarkers: pyrimidine; p-cresol; phenol; dimethyl disulfide; and dimethyl trisulfide; were identified and correlated with digestate quality. Phenol was the best predictor of TAN (R = 0.96), BOD (R = 0.92), and dissolved oxygen (DO) (R = -0.91). Phenol was also the best predictor populations of Salmonella (R = 0.95) and aerobes (R = 0.88). P-cresol was the best predictor for anaerobes (R = 0.88). The off-gas from AeD will require biofiltration or other odor control measures for a much shorter time than anaerobic decomposition. The lab-scale studies indicate that AeD burial has the potential to make burial a faster, safer, and more environmentally friendly method for emergency disposal and treatment of infectious animal carcasses and that this method should be further developed via prototype-scale field studies.

Comparison of whole genome sequencing to restriction endonuclease analysis and gel diffusion precipitin-based serotyping of Pasteurella multocida.

The gel diffusion precipitin test (GDPT) and restriction endonuclease analysis (REA) have commonly been used in the serotyping and genotyping of Pasteurella multocida. Whole genome sequencing (WGS) and single nucleotide polymorphism (SNP) analysis has become the gold standard for other organisms, offering higher resolution than previously available methods. We compared WGS to REA and GDPT on 163 isolates of P. multocida to determine if WGS produced more precise results. The isolates used represented the 16 reference serovars, isolates with REA profiles matching an attenuated fowl cholera vaccine strain, and isolates from 10 different animal species. Isolates originated from across the United States and from Chile. Identical REA profiles clustered together in the phylogenetic tree. REA profiles that differed by only a few bands had fewer SNP differences than REA profiles with more differences, as expected. The GDPT results were diverse but it was common to see a single serovar show up repeatedly within clusters. Several errors were found when examining the REA profiles. WGS was able to confirm these errors and compensate for the subjectivity in analysis of REA. Also, results of WGS and SNP analysis correlated more closely with the epidemiologic data than GDPT. In silico results were also compared to a lipopolysaccharide rapid multiplex PCR test. From the data produced in our study, WGS and SNP analysis was superior to REA and GDPT and highlighted some of the issues with the older tests.

Methicillin-Resistant Staphylococcus aureus Sequence Type (ST) 5 Isolates from Health Care and Agricultural Sources Adhere Equivalently to Human Keratinocytes.

Staphylococcus aureus is part of the nasal microbiome of many humans and has become a significant public health burden due to infections with antibiotic-resistant strains, including methicillin-resistant S. aureus (MRSA) strains. Several lineages of S. aureus, including MRSA, are found in livestock species and can be acquired by humans through contact with animals. These livestock-associated MRSA (LA-MRSA) isolates raise public health concerns because of the potential for livestock to act as reservoirs for MRSA outside the hospital setting. In the United States, swine harbor a mixed population of LA-MRSA isolates, with the sequence type 398 (ST398), ST9, and ST5 lineages being detected. LA-MRSA ST5 isolates are particularly concerning to the public health community because, unlike the isolates in the ST398 and ST9 lineages, isolates in the ST5 lineage are a significant cause of human disease in both the hospital and community settings globally. The ability of swine-associated LA-MRSA ST5 isolates to adhere to human keratinocytes in vitro was investigated, and the adherence genes harbored by these isolates were evaluated and compared to those in clinical MRSA ST5 isolates from humans with no swine contact. The two subsets of isolates adhered equivalently to human keratinocytes in vitro and contained an indistinguishable complement of adherence genes that possessed a high degree of sequence identity. Collectively, our data indicate that, unlike LA-MRSA ST398 isolates, LA-MRSA ST5 isolates do not exhibit a reduced genotypic or phenotypic capacity to adhere to human keratinocytes.IMPORTANCE Our data indicate that swine-associated livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) ST5 isolates are as capable of adhering to human skin and have the same genetic potential to adhere as clinical MRSA ST5 isolates from humans. This suggests that humans in contact with livestock have the potential to become colonized with LA-MRSA ST5 isolates; however, the genes that contribute to the persistence of S. aureus on human skin were absent in LA-MRSA ST5 isolates. The data presented here are important evidence in evaluating the potential risks that LA-MRSA ST5 isolates pose to humans who come into contact with livestock.

Draft Genome Sequences of 63 Swine-Associated Methicillin-Resistant Staphylococcus aureus Sequence Type 5 Isolates from the United States.

Methicillin-resistant Staphylococcus aureus colonizes humans and other animals such as swine. Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) sequence type 5 (ST5) isolates are a public concern due to their pathogenicity and ability to acquire mobile genetic elements. This report presents draft genome sequences for 63 LA-MRSA ST5 isolates in the United States.

Draft Genome Sequences of 14 Livestock-Associated Methicillin-Resistant Staphylococcus aureus Sequence Type 398 Isolates from Swine Farms in the United States.

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) is a bacterium carried by or obtained from swine and other livestock. The initial and predominant swine-associated LA-MRSA sequence type (ST) identified is ST398. Here, we present 14 draft genome sequences from LA-MRSA ST398 isolates found in the United States.

Draft Genome Sequences of Nine Livestock-Associated Methicillin-Resistant Staphylococcus aureus Sequence Type 5 Isolates Obtained from Humans after Short-Term Swine Contact.

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) sequence type 5 (ST5) has raised concerns surrounding the potential for these isolates to colonize or cause disease in humans with swine contact. Here, we report draft genome sequences for nine LA-MRSA ST5 isolates obtained from humans after short term swine contact.

Complete Genome Sequence of Livestock-Associated Methicillin-Resistant Staphylococcus aureus Sequence Type 398 Isolated from Swine in the United States.

Methicillin-resistant Staphylococcus aureus (MRSA) colonizes and causes disease in many animal species. Livestock-associated MRSA (LA-MRSA) isolates are represented by isolates of the sequence type 398 (ST398). These isolates are considered to be livestock adapted. This report provides the complete genome sequence of one swine-associated LA-MRSA ST398 isolate from the United States.

Complete Genome Sequence of a Livestock-Associated Methicillin-Resistant Staphylococcus aureus Sequence Type 5 Isolate from the United States.

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) may be the largest MRSA reservoir outside the hospital setting. One concern with LA-MRSA is the acquisition of novel mobile genetic elements by these isolates. Here, we report the complete genome sequence of a swine LA-MRSA sequence type 5 isolate from the United States.

Efficacy of NH3 as a secondary barrier treatment for inactivation of Salmonella Typhimurium and methicillin-resistant Staphylococcus aureus in digestate of animal carcasses: Proof-of-concept.

Managing the disposal of infectious animal carcasses from routine and catastrophic disease outbreaks is a global concern. Recent research suggests that burial in lined and aerated trenches provides the rapid pathogen containment provided by burial, while reducing air and water pollution potential and the length of time that land is taken out of agricultural production. Survival of pathogens in the digestate remains a concern, however. A potential answer is a 'dual'-barrier approach in which ammonia is used as a secondary barrier treatment to reduce the risk of pathogen contamination when trench liners ultimately leak. Results of this study showed that the minimum inhibitory concentration (MIC) of NH3 is 0.1 M (~1,468 NH3-N mg/L), and 0.5 M NH3 (~7,340 NH3-N mg/L) for ST4232 & MRSA43300, respectively at 24 h and pH = 9±0.1 and inactivation was increased by increasing NH3 concentration and/or treatment time. Results for digestate treated with NH3 were consistent with the MICs, and both pathogens were completely inactivated within 24 h.

Method for sampling and analysis of volatile biomarkers in process gas from aerobic digestion of poultry carcasses using time-weighted average SPME and GC-MS.

A passive sampling method, using retracted solid-phase microextraction (SPME) - gas chromatography-mass spectrometry and time-weighted averaging, was developed and validated for tracking marker volatile organic compounds (VOCs) emitted during aerobic digestion of biohazardous animal tissue. The retracted SPME configuration protects the fragile fiber from buffeting by the process gas stream, and it requires less equipment and is potentially more biosecure than conventional active sampling methods. VOC concentrations predicted via a model based on Fick's first law of diffusion were within 6.6-12.3% of experimentally controlled values after accounting for VOC adsorption to the SPME fiber housing. Method detection limits for five marker VOCs ranged from 0.70 to 8.44ppbv and were statistically equivalent (p>0.05) to those for active sorbent-tube-based sampling. The sampling time of 30min and fiber retraction of 5mm were found to be optimal for the tissue digestion process.

Zinc Resistance within Swine-Associated Methicillin-Resistant Staphylococcus aureus Isolates in the United States Is Associated with Multilocus Sequence Type Lineage.

Zinc resistance in livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) sequence type 398 (ST398) is primarily mediated by the czrC gene colocated with the mecA gene, encoding methicillin resistance, within the type V staphylococcal cassette chromosome mec (SCCmec) element. Because czrC and mecA are located within the same mobile genetic element, it has been suggested that the use of zinc in feed as an antidiarrheal agent has the potential to contribute to the emergence and spread of methicillin-resistant S. aureus (MRSA) in swine, through increased selection pressure to maintain the SCCmec element in isolates obtained from pigs. In this study, we report the prevalence of the czrC gene and phenotypic zinc resistance in U.S. swine-associated LA-MRSA ST5 isolates, MRSA ST5 isolates from humans with no swine contact, and U.S. swine-associated LA-MRSA ST398 isolates. We demonstrated that the prevalence of zinc resistance in U.S. swine-associated LA-MRSA ST5 isolates was significantly lower than the prevalence of zinc resistance in MRSA ST5 isolates from humans with no swine contact and swine-associated LA-MRSA ST398 isolates, as well as prevalences from previous reports describing zinc resistance in other LA-MRSA ST398 isolates. Collectively, our data suggest that selection pressure associated with zinc supplementation in feed is unlikely to have played a significant role in the emergence of LA-MRSA ST5 in the U.S. swine population. Additionally, our data indicate that zinc resistance is associated with the multilocus sequence type lineage, suggesting a potential link between the genetic lineage and the carriage of resistance determinants.IMPORTANCE Our data suggest that coselection thought to be associated with the use of zinc in feed as an antimicrobial agent is not playing a role in the emergence of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) ST5 in the U.S. swine population. Additionally, our data indicate that zinc resistance is more associated with the multilocus sequence type lineage, suggesting a potential link between the genetic lineage and the carriage of resistance markers. This information is important for public health professionals, veterinarians, producers, and consumers.

Multilaboratory Survey To Evaluate Salmonella Prevalence in Diarrheic and Nondiarrheic Dogs and Cats in the United States between 2012 and 2014.

Eleven laboratories collaborated to determine the periodic prevalence of Salmonella in a population of dogs and cats in the United States visiting veterinary clinics. Fecal samples (2,965) solicited from 11 geographically dispersed veterinary testing laboratories were collected in 36 states between January 2012 and April 2014 and tested using a harmonized method. The overall study prevalence of Salmonella in cats (3 of 542) was <1%. The prevalence in dogs (60 of 2,422) was 2.5%. Diarrhea was present in only 55% of positive dogs; however, 3.8% of the all diarrheic dogs were positive, compared with 1.8% of the nondiarrheic dogs. Salmonella-positive dogs were significantly more likely to have consumed raw food (P = 0.01), to have consumed probiotics (P = 0.002), or to have been given antibiotics (P = 0.01). Rural dogs were also more likely to be Salmonella positive than urban (P = 0.002) or suburban (P = 0.001) dogs. In the 67 isolates, 27 unique serovars were identified, with three dogs having two serovars present. Antimicrobial susceptibility testing of 66 isolates revealed that only four of the isolates were resistant to one or more antibiotics. Additional characterization of the 66 isolates was done using pulsed-field gel electrophoresis and whole-genome sequencing (WGS). Sequence data compared well to resistance phenotypic data and were submitted to the National Center for Biotechnology Information (NCBI). This study suggests an overall decline in prevalence of Salmonella-positive dogs and cats over the last decades and identifies consumption of raw food as a major risk factor for Salmonella infection. Of note is that almost half of the Salmonella-positive animals were clinically nondiarrheic.

Draft Genome Sequences of Nine Streptococcus suis Strains Isolated in the United States.

Streptococcus suis is a swine pathogen responsible for economic losses to the pig industry worldwide. Additionally, it is a zoonotic agent that can cause severe infections in those in close contact with infected pigs and/or who consume uncooked or undercooked pork products. Here, we report nine draft genome sequences of S. suis.

Comparative Prevalence of Immune Evasion Complex Genes Associated with ß-Hemolysin Converting Bacteriophages in MRSA ST5 Isolates from Swine, Swine Facilities, Humans with Swine Contact, and Humans with No Swine Contact.

Livestock associated methicillin-resistant Staphylococcus aureus (LA-MRSA) draws concern from the public health community because in some countries these organisms may represent the largest reservoir of MRSA outside hospital settings. Recent studies indicate LA-MRSA strains from swine are more genetically diverse than the first reported sequence type ST398. In the US, a diverse population of LA-MRSA is found including organisms of the ST398, ST9, and ST5 lineages. Occurrence of ST5 MRSA in swine is of particular concern since ST5 is among the most prevalent lineages causing clinical infections in humans. The prominence of ST5 in clinical disease is believed to result from acquisition of bacteriophages containing virulence or host-adapted genes including the immune-evasion cluster (IEC) genes carried by ß-hemolysin converting bacteriophages, whose absence in LA-MRSA ST398 is thought to contribute to reduced rates of human infection and transmission associated with this lineage. The goal of this study was to investigate the prevalence of IEC genes associated with ß-hemolysin converting bacteriophages in MRSA ST5 isolates obtained from agricultural sources, including swine, swine facilities, and humans with short- or long-term swine exposure. To gain a broader perspective, the prevalence of these genes in LA-MRSA ST5 strains was compared to the prevalence in clinical MRSA ST5 strains from humans with no known exposure to swine. IEC genes were not present in any of the tested MRSA ST5 strains from agricultural sources and the ß-hemolysin gene was intact in these strains, indicating the bacteriophage's absence. In contrast, the prevalence of the ß-hemolysin converting bacteriophage in MRSA ST5 strains from humans with no exposure to swine was 90.4%. The absence of ß-hemolysin converting bacteriophage in LA-MRSA ST5 isolates is consistent with previous reports evaluating ST398 strains and provides genetic evidence indicating LA-MRSA ST5 isolates may harbor a reduced capacity to cause severe disease in immunocompetent humans.

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry for rapid identification of Brachyspira species isolated from swine, including the newly described "Brachyspira hampsonii".

The Brachyspira species traditionally associated with swine dysentery and other diarrheal diseases in pigs are Brachyspira hyodysenteriae, Brachyspira pilosicoli, and, to a lesser extent, Brachyspira murdochii. "Brachyspira hampsonii" is a recently proposed novel species that causes clinical disease similar to that caused by B. hyodysenteriae. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems are increasingly available in veterinary diagnostic laboratories, are less expensive, and are faster than traditional microbiological and molecular methods for identification. Thirty-three isolates associated with Brachyspira species of importance to swine were added to an existing MALDI-TOF MS database library. In total, species included in the library were: B. hyodysenteriae, "B. hampsonii" clades I and II, Brachyspira innocens, Brachyspira intermedia, B. murdochii, and B. pilosicoli. A comparison between MALDI-TOF MS and nox sequencing was completed on 176 field isolates. Of the 176 field isolates, 174 (98.9%) matched species identification by both methods. Thirty field isolates were identified by both methods as "B. hampsonii". Twenty-seven of the 30 (90%) "B. hampsonii" field isolates matched clade designation in both assays. The nox sequencing identified 26 as "B. hampsonii" clade I and 4 as clade II. Comparatively, MALDI-TOF MS identified 25 of the 30 as "B. hampsonii" clade I and 5 as clade II. The current study indicates MALDI-TOF MS is a reliable tool for the identification of swine Brachyspira species; however, final clade designation for "B. hampsonii" may still require molecular techniques.