Swine Dysentery Research Articles

Control of swine dysentery at national level in Sweden

BackgroundSwine dysentery, caused by Brachyspira hyodysenteriae, is a severe pig disease. Resistance to tylosins is common and resistance to tiamulin has been reported since the 1990s. Still, dysentery is not notifiable to authorities. The disease therefore escapes control from an overall population perspective. In Sweden, a program that aimed to control dysentery at national level was initiated in 2020, mainly due to the unexpected diagnosis of tiamulin resistant Brachyspira hyodysenteriae in 2016.ResultsThrough joint efforts of a network including farmers, government, animal health organisations and abattoirs it was concluded that outbreaks of dysentery had taken place in 25 herds between 2016 and 2019. By 1 January 2020, nine of these herds were still not declared free from the disease. From that date, the network decided that Brachyspira hyodysenteriae was to be cultured whenever dysentery could be suspected. Thus, 148, 157 and 124 herds were scrutinised for Brachyspira hyodysenteriae in 2020, 2021 and 2022, respectively, whereof five, three and two new herds were confirmed positive. By 31 December 2022, four herds were judged as impossible to sanitise. However, they posed no problem since they were identified by the network, pigs to and from these enterprises could be transported without jeopardising other herds. When Brachyspira hyodysenteriae was diagnosed in fattening herds purchasing growers, Brachyspira hyodysenteriae could not be detected in the delivering herds. That result, together with other observations, indicated that Brachyspira hyodysenteriae ought to be regarded as ubiquitous, although at a low level in healthy pigs.ConclusionsEradication of dysentery contributed to substantial welfare and financial improvements in affected herds. Dysentery was controlled successfully at national level through the united efforts from competing stake holders, such as different abattoirs and animal health organisations. However, as Brachyspira hyodysenteriae was assumed to be ubiquitous, although at a low level in healthy pigs, the duration of the successful control of dysentery was concluded to only be transient. Without permanent monitoring for Brachyspira hyodysenteriae, the knowledge of the national status will rapidly decline to the level prior to the initiation of the control program.

First molecular detection of Brachyspira hampsonii on pig farms in Poland.

Nowadays, the three strongly beta-haemolytic spirochaetes, Brachyspira hyodysenteriae, Brachyspira suanatina and Brachyspira hampsonii are thought to be causative agents of swine dysentery, an economically devastating disease of grow-finish pigs characterised by severe mucohaemorrhagic diarrhoea. B. hyodysenteriae has been reported in most leading swine-producing regions. B. suanatina and B. hampsonii have been successfully recovered from faecal samples collected in a few countries only. Thepresent study was performed in March 2023 on faecal samples originating from nine Polish finisher farms with 6,000 to 18,000 animals in a location. Samples were obtained from 40 diarrhoeic finishers. Nucleic acid extracted from the samples was analysed using multiplex PCR for Brachyspira spp. From a total of nine sample populations examined in our study, the genetic material of B. hampsonii was identified in two. To the best of our knowledge, this is the first report on molecular detection of B. hampsonii on pig farms outside North America, Belgium and Germany. Our research highlights the need for increased focus directed on laboratory testing strategies, the lack of which may perplex swine practitioners and severely hinder a definite diagnosis.

New insights into swine dysentery: faecal shedding, macro and microscopic lesions and biomarkers in early and acute stages of Brachyspira hyodysenteriae infection

BackgroundSwine dysentery (SD) is a severe mucohaemorrhagic colitis in pigs caused classically by Brachyspira hyodysenteriae. Although several aspects of B. hyodysenteriae infection dynamic are already described, further research in the early stage of this infection is required. In this study, 7-week-old pigs were orally challenged with B. hyodysenteriae to obtain information about faecal shedding, macro and microscopic intestinal lesions and serum acute phase proteins in pigs at the onset of B. hyodysenteriae shedding (early infection group, n = 8), in pigs with mucohaemorrhagic diarrhoea (acute infection group, n = 8) and in non-infected controls (n = 16).ResultsFirst B. hyodysenteriae detection by q-PCR and first loose stools with blood and mucus occurred both at 8 days post-inoculation. The lapse between a positive q-PCR and observation of mucohaemorrhagic diarrhoea ranged from 0 to 3 days, except in a single pig in which this period lasted 5 days. Macroscopic lesions were observed in the large intestine from both infected groups although more frequent and severe in acute infection group. Microscopic observation of the apex mucosa revealed that in early infection only higher ulceration values were observed compared to healthy controls. In contrast, the acute infection group exhibited higher ulceration, neutrophils infiltration and increased mucosal thickness compared to the other two groups. Among the serum biomarkers tested, only haptoglobin, C-reactive protein, and creatine kinase showed a significant increase in pigs in the acute infection period compared to controls, whereas haptoglobin was the only factor with a significant increase at the early infection compared to non-infected animals.ConclusionsThis study provides new insights about SD and remarks the complex and limited options to perform an early detection of infected animals beyond PCR diagnosis.

In vitro susceptibility of Brachyspira hyodysenteriae strains isolated in pigs in northern Italy between 2005 and 2022

Pleuromutilins (tiamulin and valnemulin) are often used to treat swine dysentery due to recurrent resistance to macrolides and lincosamides. Recently, reduced susceptibility of B. hyodysenteriae to pleuromutilin has been reported. 536 strains of B. hyodysenteriae were isolated from symptomatic pigs weighing 30–150 kg in northern Italy between 2005 and 2022. B. hyodysenteriae was isolated by standard methods and confirmed by PCR. The minimum inhibitory concentration (MIC) to doxycycline, lincomycin, tiamulin, tylosin, tylvalosine and valnemulin was evaluated according to CLSI procedures and MIC data were reported as MIC 50 and MIC 90. The temporal trend of the MIC values was evaluated by dividing the data into two groups (2005–2013 and 2014–2022). Comparison of the distribution in frequency classes in the two periods was performed using Pearson's chi-squared test (p < 0.01).MIC 50 was close to the highest values tested for lincomycin and tylosin, while MIC 90 was close to the highest values tested for all antibiotics. 71.7% of the strains were susceptible to tylvalosin, while 75%–80.4% had reduced susceptibility to valnemulin and tiamulin, respectively. The difference in the distribution of MIC classes was statistically significant in the two periods for doxycycline, tiamulin, tylvalosin and valnemulin, and more MIC classes above the epidemiological cut-off were observed in 2014–2022 compared with 2005–2013. The evaluation of the trends during the period considered shows a decreasing rate of wild-type strains with MIC values below the epidemiological cut-off over time and confirms the presence of resistant strains in northern Italy.

T-independent B-cell effect of agents associated with swine grower-finisher diarrhea.

Swine dysentery, spirochetal colitis, and salmonellosis are production-limiting enteric diseases of global importance to the swine industry. Despite decades of efforts, mitigation of these diseases still relies on antibiotic therapy. A common knowledge gap among the 3 agents is the early B-cell response to infection in pigs. Thus, this study aimed to characterize the porcine B-cell response to Brachyspira hyodysenteriae, Brachyspira hampsonii (virulent and avirulent strains), Brachyspira pilosicoli, and Salmonella Typhimurium, the agents of the syndromes mentioned above. Immortalized porcine B-cell line derived from a crossbred pig with lymphoma were co-incubated for 8h with each pathogen, as well as E. coli lipopolysaccharide (LPS) and a sham-inoculum (n = 3/treatment). B-cell viability following treatments was evaluated using trypan blue, and the expression levels of B-cell activation-related genes was profiled using reverse transcription quantitative PCR. Only S. Typhimurium and LPS led to increased B-cell mortality. B. pilosicoli downregulated B-lymphocyte antigen (CD19), spleen associated tyrosine Kinase (syk), tyrosine-protein kinase (lyn), and Tumour Necrosis Factor alpha (TNF-α), and elicited no change in immunoglobulin-associated beta (CD79b) and swine leukocyte antigen class II (SLA-DRA) expression levels, when compared to the sham-inoculated group. In contrast, all other treatments significantly upregulated CD79b and stimulated responses in other B-cell downstream genes. These findings suggest that B. pilosicoli does not elicit an immediate T-independent B-cell response, nor does it trigger antigen-presenting mechanisms. All other agents activated at least one trigger within the T-independent pathways, as well as peptide antigen presenting mechanisms. Future research is warranted to verify these findings in vivo.

Antimicrobial susceptibility of U.S. porcine Brachyspira isolates and genetic diversity of B. hyodysenteriae by multilocus sequence typing.

Swine dysentery, caused by Brachyspira hyodysenteriae and the newly recognized Brachyspira hampsonii in grower-finisher pigs, is a substantial economic burden in many swine-rearing countries. Antimicrobial therapy is the only commercially available measure to control and prevent Brachyspira-related colitis. However, data on antimicrobial susceptibility trends and genetic diversity of Brachyspira species from North America is limited. We evaluated the antimicrobial susceptibility profiles of U.S. Brachyspira isolates recovered between 2013 and 2022 to tiamulin, tylvalosin, lincomycin, doxycycline, bacitracin, and tylosin. In addition, we performed multilocus sequence typing (MLST) on 64 B. hyodysenteriae isolates. Overall, no distinct alterations in the susceptibility patterns over time were observed among Brachyspira species. However, resistance to the commonly used antimicrobials was seen sporadically with a higher resistance frequency to tylosin compared to other tested drugs. B. hampsonii was more susceptible to the tested drugs than B. hyodysenteriae and B. pilosicoli. MLST revealed 16 different sequence types (STs) among the 64 B. hyodysenteriae isolates tested, of which 5 STs were previously known, whereas 11 were novel. Most isolates belonged to the known STs: ST93 (n = 32) and ST107 (n = 13). Our findings indicate an overall low prevalence of resistance to clinically important antimicrobials other than tylosin and bacitracin, and high genetic diversity among the clinical Brachyspira isolates from pigs in the United States during the past decade. Further molecular, epidemiologic, and surveillance studies are needed to better understand the infection dynamics of Brachyspira on swine farms and to help develop effective control measures.

Characterization of the bacterial fecal microbiota composition of pigs preceding the clinical signs of swine dysentery.

Swine dysentery (SD) is a worldwide production-limiting disease of growing-finishing pigs in commercial farms. The importance of the large intestinal microbiota in the swine dysentery pathogenesis has been established, but not well characterized. The objective of this study was to characterize the fecal bacterial microbiota of pigs immediately prior to developing clinical signs of swine dysentery. A total of 60 fecal samples were collected from 15 pigs with SD. Sampling times included a time point prior to SD (d0, n=15), 2 days before mucohaemorrhagic diarrhea was observed (d-2SD, n=15), 1 day before mucohaemorrhagic diarrhea was observed (d-1SD, n=15), and the day when pigs developed mucohemorragic diarrhea (MHD, n=15). Sequencing of cpn60 amplicons was used to profile the microbiome, and analyses were performed on QIIME2. Increased Chao1 index in d-1SD and MHD samples when compared to the d0 was the only change observed in alpha diversity. No differences between sampling times on beta diversity (Bray-Curtis dissimilarity) were found. Although a small sample size was investigated, differential abundance analysis revealed that Alistipes dispar and Parabacteroides gordonii were increased in MHD fecal samples when compared to d-2SD and d-1SD. It is suggested that these taxa may play a role in the pathogenesis of SD, which is known to require the presence of Brachyspira spp. and an anaerobe for severe disease development.

Metagenomic analysis fecal microbiota of dysentery-like diarrhoea in a pig farm using next-generation sequencing.

Porcine enteric diseases including swine dysentery involves a wide range of possible aetiologies and seriously damages the intestine of pigs of all ages. Metagenomic next-generation sequencing is commonly used in research for detecting and analyzing pathogens. In this study, the feces of pigs from a commercial swine farm with dysentery-like diarrhea was collected and used for microbiota analysis by next-generation sequencing. While Brachyspira spp. was not detected in diarrheal pig fecal samples, indicating that the disease was not swine dysentery. The quantity of microbial population was extremely lowered, and the bacterial composition was altered with a reduction in the relative abundance of the probiotics organisms, Firmicutes and Bacteroidetes, with an increase in pathogens like Fusobacterium and Proteobacteria, in which the specific bacteria were identified at species-level. Viral pathogens, porcine circovirus type 2, porcine lymphotropic herpesviruses 1, and porcine mastadenovirus A were also detected at pretty low levels. Carbohydrate-active enzymes (CAZy) analysis indicated that the constitute of Firmicutes and Bacteroidete were also changed. Further, the Kyoto Encyclopedia of Genes and Genomes (KEGG) alignment analysis indicated that the microbiota of diarrheal pigs had a lower ability in utilizing energy sources but were enriched in multi-drug resistance pathways. Comprehensive Antibiotic Resistance Database (CARD) and Virulence Factors of Pathogenic Bacteria (VFDB) analysis indicated that genes for elfamycin and sulfonamide resistance and the iron uptake system were enriched in diarrheal pigs. This revealed potential bacterial infection and can guide antibiotic selection for treating dysentery. Overall, our data suggested that alterations in both the population and functional attributes of microbiota in diarrheal pigs with decreased probiotic and increased pathogenic microorganisms. These results will help elucidate the mechanism of dysentery-like diarrhea and the development of approaches to control the disease.

First molecular detection of Brachyspira suanatina on pig farms in Poland.

Prior to the 2000s, swine dysentery was considered to be caused only by Brachyspira hyodysenteriae with contributing commensal intestinal anaerobes. Nowadays, it is known that the disease is caused by three strongly beta-haemolytic species of the anaerobic spirochaetal genus Brachyspira, i.e. B. hyodysenteriae and newly emerged B. hampsonii and B. suanatina. The present investigation was carried out in November 2022 on nine Polish high-performing finisher pig farms. At every location one fresh pooled faecal sample was collected from 40 randomly selected pigs of between 60 and 110 kg live weight. Nucleic acid extracted from each pooled faecal sample was analysed by an in-house multiplex PCR for Brachyspira spp., which is capable of confirming the Brachyspira genus and detecting and differentiating Brachyspira species. From a total of nine samples examined, the genetic material of B. suanatina was detected in seven. Non-pathogenic/questionably pathogenic Brachyspira spp. were found in six samples. To the best of our knowledge, this is the first report on the identification of B. suanatina in pigs outside Scandinavia, Germany and the United Kingdom. Our research not only provides valuable epidemiological data on B. suanatina infection in Europe but also highlights both the importance of modern laboratory diagnostics and the need for thorough investigation across regions, including retrospective studies.

Synergic Effect of Brachyspira hyodysenteriae and Lawsonia intracellularis Coinfection: Anatomopathological and Microbiome Evaluation.

Brachyspira hyodysenteriae and Lawsonia intracellularis coinfection has been observed in the diagnostic routine; however, no studies have evaluated their interaction. This study aimed to characterize lesions and possible synergisms in experimentally infected pigs. Four groups of piglets, coinfection (CO), B. hyodysenteriae (BRA), L. intracellularis (LAW), and negative control (NEG), were used. Clinical signals were evaluated, and fecal samples were collected for qPCR. At 21 days post infection (dpi), all animals were euthanized. Gross lesions, bacterial isolation, histopathology, immunohistochemistry, and fecal microbiome analyses were performed. Diarrhea started at 12 dpi, affecting 11/12 pigs in the CO group and 5/11 pigs in the BRA group. Histopathological lesions were significantly more severe in the CO than the other groups. B. hyodysenteriae was isolated from 11/12 pigs in CO and 5/11 BRA groups. Pigs started shedding L. intracellularis at 3 dpi, and all inoculated pigs tested positive on day 21. A total of 10/12 CO and 7/11 BRA animals tested positive for B. hyodysenteriae by qPCR. A relatively low abundance of microbiota was observed in the CO group. Clinical signs and macroscopic and microscopic lesions were significantly more severe in the CO group compared to the other groups. The presence of L. intracellularis in the CO group increased the severity of swine dysentery.

Nosological profile and classification of infectious diseases of farm animals in the territory of the Pepublic of Sakha (Yakutia)

The results of the classification of infectious diseases of farm animals spread on the territory of the Republic of Sakha (Yakutia) for the period 2001-2021 are presented. It was found that there were 42 nosological units of animal diseases involved in the epizootic process on the territory of Yakutia. Infections in cattle are caused by 18 types of pathogens, horses - 12, reindeer - 4, pigs - 10, birds - 11, small ruminants - 3, fur-bearing animals - 7, other types of animals - 9. According to the OIE classification, 10 nosoforms of diseases are common in Yakutia: rabies, brucellosis, viral hemorrhagic septicemia, equine infectious anemia, avian infectious bronchitis, infectious bovine rhinotracheitis, paratuberculosis, pasteurellosis and tuberculosis. Diseases not on the OIE list, including Marek’s disease, swine dysentery, colibacillosis, parvoviral enteritis, salmonellosis, diamond-skin disease, blackleg were registered. A number of nosoforms common in Yakutia belong to particularly dangerous diseases for which restrictive measures (quarantine) may be imposed. These include actinomycosis, rabies, brucellosis, infectious equine anemia, avian infectious bronchitis, infectious bovine rhinotracheitis, bovine leukosis, leptospirosis, paratuberculosis, diamond-skin disease, tuberculosis and canine distemper virus. A number of diseases (Marek’s disease, carnivorous infectious hepatitis, bovine leukosis, horse strangles, avian ornithosis, egg drop syndrome, diamond-skin disease, and blackleg) are peculiar to only one species of animals. Many infections are characterized by the persistence of the pathogen in different species of farm and small domestic animals, birds and wildlife, which determines the interspecies degree of epizootic danger of these infections. Infections of bacterial etiology (bacterioses) prevail in Yakutia with 21 nosoforms. Viral diseases (viroses) are represented by 15 nosoforms, mycoplasmosis and chlamydia - one each. The most numerous infections are those with the main alimentary, or fecal-oral, route (22 diseases). Ten infections, including infectious atrophic rhinitis, infectious bovine rhinotracheitis, distemper, are classified as airborne or respiratory (major) pathogen transmission. Transmissive carry-over (by vectors) is characterized by actinomycosis and infectious equine anemia; through the outer covers (without the participation of vectors) - 5 diseases, including balanoposthitis and rabies. The majority of the infections are characterized by one or more additional mechanisms of transmission (rabies, Marek’s disease, bovine leukemia, diamond-skin disease, etc.) in addition to the main mechanism of transmission of the pathogen. According to the source of the causative agent for all diseases (ktenonoses) they are domestic animals, but for such infections as rabies, Marek’s disease, viral hemorrhagic septicemia, leptospirosis and others, along with domestic animals, wild animals are an additional source (ktenotherionoses). In terms of susceptibility, most infections (30) belong to zoonoses peculiar only to animals; the remaining 12 belong to the more dangerous zooanthroponoses common to animals and humans.

Improving the consistency of experimental swine dysentery inoculation strategies

Swine dysentery (SD) caused by pathogenic Brachyspira spp. is an economic challenge for the swine industry. In research settings, experimental reproduction of swine dysentery typically relies on intragastric inoculation which has shown variable success. This project aimed to improve the consistency of the experimental inoculation protocol used for swine dysentery in our laboratory. Over six experiments, we evaluated the influence of group housing in inoculated pigs using a frozen-thawed broth culture of strongly hemolytic B. hyodysenteriae strain D19 (Trial A), compared the relative virulence of B. hyodysenteriae strains D19 and G44 (Trial B), compared inoculum volumes (50 mL vs 100 mL) for G44 and B. hampsonii 30446 (Trial C), and performed three independent trials evaluating intragastric inoculation using different oral inoculation methods: oral feed balls (Trial D), and oral syringe bolus of 100 mL (Trial E) or 300 mL (Trial F). Intragastric inoculation with a fresh broth culture of B. hyodysenteriae strain G44 resulted in a shorter incubation period and a higher proportionate duration of mucohemorrhagic diarrhea (MMHD) compared to D19. Intragastric inoculation with either 50 or 100 mL of B. hampsonii 30446 or B. hyodysenteriae (G44) were statistically equivalent. Oral inoculation with 100 mL or 300 mL also yielded similar results to intragastric inoculation but was more expensive due to the additional work and supplies associated with syringe training. Our future research will use intragastric inoculation with 100 mL of a fresh broth culture containing B. hyodysenteriae strain G44 as it yields a high incidence of mucohaemorrhagic diarrhea with a reasonable cost.

The Impact of PRRS Eradication Program on the Production Parameters of the Hungarian Swine Sector.

The Hungarian national eradication program of PRRS was successfully completed between 2014 and 2022. There were doubts about the efficiency of the eradication program in Hungary from the beginning to the tune that it might only be carried out efficiently through depopulation-repopulation of the infected herds, which is a very costly procedure. In our study, we investigated the impact of the depopulation-repopulation procedure, which played a prominent role in the PRRS eradication program on the productivity of the Hungarian swine sector-namely, on the number of slaughter pigs per sow per year and the total live slaughter weight per sow per year. Since 2014, we monitored the evolution of the PRRS eradication through the depopulation-repopulation approach on the large-scale breeding herds in Hungary. Most producers replaced their herds with animals that were free of PRRS and other infectious diseases (mycoplasmosis, actinobacillosis, swine dysentery, atrophic rhinitis, etc.). On this basis, we evaluated the change in the number of slaughter pigs per sow per year as a consequence of depopulation-repopulation of the herds being carried out. In the statistical analysis linear regression was used. The results of our study demonstrate that the PRRS eradication program with the herd depopulation-repopulation approach led to a considerable improvement of the productivity of Hungarian pig farming. This result also demonstrates that, independent of the PRRS eradication, it is still necessary to consider investments into the individual production units to increase efficiency, and to carry out herd depopulation-repopulation in cases where the current genetics limits improvements in productivity.

The Incidence of Swine Dysentery in Pigs Can Be Reduced by Feeding Diets That Limit the Amount of Fermentable Substrate Entering the Large Intestine

Two experiments were conducted to test the hypothesis that feeding diets which limit the amount of fermentable substrate entering the large intestine would protect pigs against experimental infection with Serpulina hyodysenteriae, the causative agent of swine dysentery. Experiment 1 examined the effect of grain processing (hammer milling vs. steam flaking) and grain type (barley, groats, corn, sorghum and wheat) on indices of fermentation in the large intestine and the incidence of swine dysentery. Experiment 2 examined the role of five diets, steam-flaked corn, steam-flaked sorghum, hammer-milled wheat, extruded wheat and cooked white rice, on these same measures. All diets contained an animal protein supplement and no antibiotics. Pigs fed diets based on steam-flaked corn and steam-flaked sorghum had a lower incidence of disease (11–33%) than pigs fed diets based on other grains (75–100%). Pigs fed the diet based on cooked white rice were fully protected against swine dysentery. Both the soluble non-starch polysaccharide (NSP) concentration and the total NSP concentration of the diets explained a significant proportion of the variation in swine dysentery (R2 = 0.56, P = 0.016, and R2 = 0.71, P = 0.002, respectively), such that pigs eating diets containing <1.0 g/100 g soluble NSP showed reduced disease. However, pigs fed corn, sorghum and steam-flaked sorghum (Experiment 2), which contained only 0.4–0.5 g/100 g soluble NSP, still had a high incidence of disease (>50%). This was attributable to a higher level of resistant starch present in these grains. These data provide evidence that the expression of swine dysentery is associated with an increased concentration of fermentable substrate entering the large intestine.

Prevalence and risk factors of Brachyspira spp. in pig herds with a history of diarrhoea in six European countries

Swine dysentery and porcine intestinal spirochaetosis caused by Brachyspira (B.) hyodysenteriae and B. pilosicoli, respectively, are important diseases in swine production worldwide. The aim of this study was to assess the prevalence of both pathogens in farms with a history of diarrhoea within the last 12 months in Denmark, France, Germany, the Netherlands, Spain, and United Kingdom. In addition, risk factors for their prevalence and correlations between presence of different Brachyspira spp. and Lawsonia intracellularis were investigated. Therefore, faecal samples of 6355 nursery to finishing pigs out of 144 herds were sampled in 2017/2018 during a prevalence study on Lawsonia intracellularis, followed by polymerase chain reaction analysis for Brachyspira spp. detection. Herd prevalence differed significantly between countries, from 4.2% to 45.8% for B. hyodysenteriae and 8.3–87.5% for B. pilosicoli, respectively (p < 0.01). For the within-herd prevalence (in positive herds), these values ranged from 2.2% to 27.0% for B. hyodysenteriae and 3.3–50.8% for B. pilosicoli. Mixed infections occurred in 34.1% and 58.7% of B. hyodysenteriae positive samples with Lawsonia intracellularis or B. pilosicoli, respectively. In 43.2% of B. pilosicoli positive samples, Lawsonia intracellularis was detected simultaneously. Overall, nursery pigs were significantly less often positive for one of the pathogens than growing or finishing pigs (p < 0.001). The absence of gastrointestinal problems like diarrhoea, routine use of antimicrobials and well performed biosecurity measures were some of the factors associated with lower detection rate of Brachyspira spp. Surprisingly, deworming of different age categories also showed associations with the detection of Brachyspira spp. which, however, were not always equally directed, and therefore require further investigations. The only risk factor significant for both Brachyspira spp. was the median number of ≥ 30 nursery pigs per pen after weaning, compared to smaller group sizes. Both pathogens were detected with varying frequency between the six European countries. This should be considered in the probability of disease and in case of transnational transport, to prevent spread of pathogens. In addition, the frequent presence of mixed infections in some countries should be taken into account in diagnostics. The most important protective factors against Brachyspira spp. presence on farm are biosecurity measures, while potential new factors such as deworming still require further investigation.

Porcine intestinal glycosphingolipids recognized by Brachyspira hyodysenteriae.

Swine dysentery caused by Brachyspira hyodysenteriae is a disease present worldwide with an important economic impact on the farming business, resulting in an increased use of antibiotics. In the present study, we investigated the binding of B. hyodysenteriae to glycosphingolipids from porcine small intestinal epithelium in order to determine the glycosphingolipids involved in B. hyodysenteriae adhesion. Specific interactions between B. hyodysenteriae and two non-acid glycosphingolipids were obtained. These binding-active glycosphingolipids, were characterized by mass spectrometry as lactotetraosylceramide (Galβ3GlcNAcβ3Galβ4Glcβ1Cer) and the B5 glycosphingolipid (Galα3Galβ4GlcNAcβ3Galβ4Glcβ1Cer). Comparative binding studies using structurally related reference glycosphingolipids showed that B. hyodysenteriae binding to lactotetraosylceramide required an unsubstituted terminal Galβ3GlcNAc sequence, while for binding to the B5 pentaosylceramide the terminal Galα3Galβ4GlcNAc sequence is the minimum element recognized by the bacteria. Binding of Griffonia simplicifolia IB4 lectin to pig colon tissue sections from healthy control pig and B. hyodysenteriae infected pigs showed that in the healthy pigs the Galα3Gal epitope was mainly present in the lamina propria. In contrast, in four out of five pigs with swine dysentery there was an increased expression of Galα3Gal in the goblet cells and in the colonic crypts, where B. hyodysenteriae also was present. The one pig that had recovered by the time of necropsy had the Galα3Gal epitope only in the lamina propria. These data are consistent with a model where a transient increase in the carbohydrate sequence recognized by the bacteria occur in colonic mucins during B. hyodysenteriae infection, suggesting that the mucins may act as decoys contributing to clearance of the infection. These findings may lead to novel strategies for treatment of B. hyodysenteriae induced swine dysentery.

Acute infection with Brachyspira hyodysenteriae affects mucin expression, glycosylation, and fecal MUC5AC.

Infection with strongly β-hemolytic strains of Brachyspira hyodysenteriae leads to swine dysentery (SD), a production-limiting disease that causes mucohemorrhagic diarrhea and typhlocolitis in pigs. This pathogen has strong chemotactic activity toward mucin, and infected pigs often have a disorganized mucus layer and marked de novo expression of MUC5AC, which is not constitutively expressed in the colon. It has been shown that fucose is chemoattractant for B. hyodysenteriae, and a highly fermentable fiber diet can mitigate and delay the onset of SD. We used lectins targeting sialic acids in α-2,6 or α-2,3 linkages, N-acetylglucosamine (GlcNAc), α-linked L-fucose, and an immunohistochemical stain targeting N-glycolylneuraminic acid (NeuGc) to investigate the local expression of these mucin glycans in colonic tissues of pigs with acute SD. We used a commercial enzyme-linked immunosorbent assay (ELISA) to quantify fecal MUC5AC in infected pigs and assess its potential as a diagnostic monitoring tool and RNA in situ hybridization to detect IL-17A in the colonic mucosa. Colonic mucin glycosylation during SD has an overall increase in fucose, a spatially different distribution of GlcNAc with more expression within the crypt lumens of the upper colonic mucosa, and decreased expression or a decreased trend of sialic acids in α-2,6 or α-2,3 linkages, and NeuGc compared to the controls. The degree of increased fucosylation was less in the colonic mucosa of pigs with SD and fed the highly fermentable fiber diet. There was a significant increase in MUC5AC in fecal and colonic samples of pigs with SD at the endpoint compared to the controls, but the predictive value for disease progression was limited. Fucosylation and the impact of dietary fiber may play important roles in the pathogenesis of SD. The lack of predictive value for fecal MUC5AC quantification by ELISA is possibly due to the presence of other non-colonic sources of MUC5AC in the feces. The moderate correlation between IL-17A, neutrophils and MUC5AC confirms its immunoregulatory and mucin stimulatory role. Our study characterizes local alteration of mucin glycosylation in the colonic mucosa of pigs with SD after B. hyodysenteriae infection and may provide insight into host-pathogen interaction.

Predictive Power of Long-Read Whole-Genome Sequencing for Rapid Diagnostics of Multidrug-Resistant Brachyspira hyodysenteriae Strains.

Infections with Brachyspira hyodysenteriae, the etiological agent of swine dysentery, result in major economic losses in the pig industry worldwide. Even though microbial differentiation of various Brachyspira species can be obtained via PCR, no quick diagnostics for antimicrobial susceptibility testing are in place, which is mainly due to the time-consuming (4 to 7 days) anaerobic growth requirements of these organisms. Veterinarians often rely on a clinical diagnosis for initiating antimicrobial treatment. These treatments are not always effective, which may be due to high levels of acquired resistance in B. hyodysenteriae field isolates. By using long-read-only whole-genome sequencing and a custom-trained Bonito base-calling model, 81 complete B. hyodysenteriae genomes with median Q51 scores and 99% completeness were obtained from 86 field strains. This allowed the assessment of the predictive potential of genetic markers in relation to the observed acquired resistance phenotypes obtained via agar dilution susceptibility testing. Multidrug resistance was observed in 77% and 21% of the tested strains based on epidemiological cutoff and clinical breakpoint values, respectively. The predictive power of genetic hallmarks (genes and/or gene mutations) for antimicrobial susceptibility testing was promising. Sensitivity and specificity for tiamulin [tva(A) and 50SL3N148S, 99% and 67%], valnemulin [tva(A), 97% and 92%), lincomycin (23SA2153T/G and lnuC, 94% and 100%), tylvalosin (23SA2153T/G, 99% and 93%), and doxycycline (16SG1026C, 93% and 87%) were determined. The predictive power of these genetic hallmarks is promising for use in sequencing-based workflows to speed up swine dysentery diagnostics in veterinary medicine and determine proper antimicrobial use. IMPORTANCE Diagnostics for swine dysentery rely on the identification of Brachyspira species using molecular techniques. Nevertheless, no quick diagnostic tools are available for antimicrobial susceptibility testing due to extended growth requirements (7 to 14 days). To enable practitioners to tailor antimicrobial treatment to specific strains, long-read sequencing-based methods are expected to lead to rapid methods in the future. Nevertheless, their potential implementation should be validated extensively. This mainly implies assessing sequencing accuracy and the predictive power of genetic hallmarks in relation to their observed (multi)resistance phenotypes.

ИНВАЗИВНОСТЬ BR. HYODYSENTERIAE И ДИНАМИКА ГИСТО-УЛЬТРАСТРУКТУРНЫХ ИЗМЕНЕНИЙ В СЛИЗИСТОЙ ОБОЛОЧКЕ ТОЛСТОГО КИШЕЧНИКА ПРИ ДИЗЕНТЕРИИ СВИНЕЙ

The paper discusses the urgent problems associated with the effect of β-hemolytic Br. hyodysenteriae on pathological processes in the large intestine. For establishing histological and ultrastructural changes in the large intestine, the study used the material from the slaughtered animals. The intestinal mucosa was examined according to generally accepted histological and ultrastructural methods. The tryptic soyabean casein blood agar was used for isolation of Br. hyodysenteriae under anaerobic conditions. Histological and ultrastructural examination of the intestinal mucosa in pigs with swine dysentery revealed catarrhal hemorrhagic inflammation with focal necrosis of the epithelium, edema of lamina propria. The earliest changes in colonocytes include destruction of microvilli, which is a morphological expression of membrane indigestion and malabsorption. The damaging effect of pathogens involves desquamated colonocytes, bare stroma, circulatory abnormalities, cell element proliferation. Mucoid and fibrinoid swelling of collagen fibres, fragmentation, coarsening of elastic and reticular fibres, proliferation of fibroblasts and histiocytes are observed in the connective tissue stroma of the intestine. The study established the effect of Br.hyodysenteriae on structural changes in the large intestine. The findings should be applied to the areas of a differential diagnosis and swine dysentery control measures.

In vitro evaluation of gentamicin activity against Spanish field isolates of Brachyspira hyodysenteriae

BackgroundThe treatment of swine dysentery (SD) has become constrained in recent years due to the limited availability of effective drugs combined with a rise in antimicrobial resistance. Gentamicin, an aminoglycoside antibiotic, is authorised for the control of this disease in several European countries but has not been extensively used so far. In this study, the in vitro susceptibility of 56 Brachyspira hyodysenteriae field isolates was evaluated against gentamicin using a broth microdilution test. The molecular basis of decreased susceptibility to gentamicin was also investigated by sequencing the 16S rRNA gene and phylogenetic relatedness by multiple-locus variable number tandem-repeat analysis (MLVA).ResultsMost B. hyodysenteriae isolates presented low minimum inhibitory concentration (MIC) values to gentamicin, with a mode of 2 µg/mL, a median or MIC50 of 4 µg/mL and percentile 90 or MIC90 of 16 µg/mL. The distribution of these values over the period studied (2011–2019) did not show a tendency towards the development of resistance to gentamicin. Differences in susceptibility among isolates could be explained by two point-mutations in the 16S rRNA gene, C990T and A1185G, which were only present in isolates with high MICs. These isolates were typed in three different MLVA clusters. Analyses of co-resistance between gentamicin and antimicrobials commonly used for the treatment of SD revealed that resistance to tiamulin and valnemulin was associated with low MICs for gentamicin.ConclusionsThe results provide an accurate characterisation of antimicrobial sensitivity to gentamicin and possible mechanisms of resistance in Spanish B. hyodysenteriae isolates. These findings allow us to propose gentamicin as an alternative in the antibiotic management of SD, particularly in outbreaks caused by pleuromutilin resistant isolates.