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Daisy Vanrompay - One of the best experts on this subject based on the ideXlab platform.
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Co-Occurrence of Chlamydia suis DNA and Chlamydia suis-Specific Antibodies in the Human Eye
Vector borne and zoonotic diseases (Larchmont N.Y.), 2018Co-Authors: Evelien Kieckens, Servaas A Morré, Laura Van Den Broeck, Matthias Van Gils, Daisy VanrompayAbstract:Chlamydia suis is a swine pathogen that causes economic losses due to reproductive failure. Recently, C. suis has been detected in human eyes. However, knowledge of the zoonotic potential is still limited. C. suis infections in swine could present a risk for public health because (1) tetracycline-resistant C. suis strains are emerging in the pork industry, (2) tetracycline resistance gene transfers in vitro from C. suis to the human pathogen Chlamydia trachomatis and as previously demonstrated, (3) C. suis and C. trachomatis can be both present in the human eye. Pig farmers were sampled during a seminar in West-Flanders. Conjunctival swabs for detection of C. suis and C. trachomatis and for the detection of mucosal antibodies against C. suis and C. trachomatis were collected. The farmers completed a questionnaire designed to assess information on the following: (1) the health status of their pigs, (2) administration of veterinary drugs, (3) their professional and nonprofessional activities, (4) general health status, (5) smoking habits, (6) use of medication, (7) allergies, and (8) clinical signs/history. Thirty-three on 40 (82.5%) farmers participated. None of the conjunctival swabs contained C. trachomatis DNA and mucosal antibodies against C. trachomatis were not detected. Six of 33 (18.2%) farmers had C. suis DNA in their eyes and 22 of 33 (67%) swabs contained C. suis-specific mucosal antibodies. The older the farmer, higher the chance of finding C. suis antibodies in the eye. There was a significant correlation between the presence of conjunctivitis in the pigs and the occurrence of C. suis DNA in the eye of their owner. This study shows that C. suis may transfer from pigs to the human eye as specific mucosal antibodies were detected in conjunctivae of pig farmers. Veterinarians, general practitioners, and occupational physicians should be aware of the zoonotic potential of C. suis.
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Development of a Chlamydia suis‐specific antibody enzyme‐linked immunosorbent assay based on the use of a B‐cell epitope of the polymorphic membrane protein C
Transboundary and emerging diseases, 2018Co-Authors: K. De Puysseleyr, Garry S. A. Myers, Evelien Kieckens, L. De Puysseleyr, H. Van Den Wyngaert, B. Ahmed, S. Van Lent, Heather Huot Creasy, Daisy VanrompayAbstract:Chlamydia suis infections lead to economic loss in the pork industry. Chlamydia suis infections could be successfully treated with tetracyclines until the appearance of a tetracycline resistant phenotype, which was acquired via horizontal gene transfer of the tet(C) gene. Given the importance of C. suis as a swine pathogen and as a recently emerged tetracycline resistant pathogen with zoonotic potential, our aim was to develop a sensitive C. suis-specific antibody ELISA based on the polymorphic membrane proteins (Pmps). Chlamydia Pmps are important virulence factors and candidate antigens for serodiagnosis. We identified nine Pmps (PmpA to I) in C. suis strain MD56 using a recently developed Hidden-Markov model. PmpC was the most promising candidate for the development of a C. suis-specific antibody ELISA as the protein was absent in C. abortus, C. pecorum and C. psittaci which also infect pigs and as the protein contained C. suis-specific amino acid regions, absent in C. trachomatis PmpC. We identified an immunodominant B-cell epitope in C. suis PmpC using experimental porcine sera. The sensitivity and specificity of the PmpC ELISA was compared to the complement fixation test (CFT) and to a recombinant MOMP ELISA using experimental sera. The PmpC ELISA detected all positive control sera and was in contrast to CFT and the rMOMP ELISA 100% C. suis specific as positive control sera against other Chlamydia species did not react in the PmpC ELISA. The test was successfully validated using slaughterhouse sera and sera from clinically affected pigs. The PmpC ELISA could assist in diminishing the spread of C. suis infections in the pork industry.
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quantifying the growth of Chlamydia suis in cell culture using high content microscopy
Microscopy Research and Technique, 2017Co-Authors: Leentje De Puysseleyr, Daisy Vanrompay, Kristien De Puysseleyr, Winnok H. De VosAbstract:The porcine pathogen Chlamydia suis is widespread in pig farming. Isolation of Chlamydia suis in cell culture is crucial for the generation and characterization of new isolates. However, isolation of Chlamydia suis strains from field samples is fastidious. Therefore, we exploited high-content microscopy to quantify the growth of Chlamydia suis strains in different cell lines. We found that the cell line yielding optimal propagation of Chlamydia suis differed among isolates, and we identified cell lines outperforming those routinely used for Chlamydial isolation. We conclude that adaptation of the propagation procedure to the origin of the putative field isolate is highly recommended to improve the recovery rate.
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Quantifying the growth of Chlamydia suis in cell culture using high‐content microscopy
Microscopy research and technique, 2016Co-Authors: Leentje De Puysseleyr, Daisy Vanrompay, Kristien De Puysseleyr, Winnok H. De VosAbstract:The porcine pathogen Chlamydia suis is widespread in pig farming. Isolation of Chlamydia suis in cell culture is crucial for the generation and characterization of new isolates. However, isolation of Chlamydia suis strains from field samples is fastidious. Therefore, we exploited high-content microscopy to quantify the growth of Chlamydia suis strains in different cell lines. We found that the cell line yielding optimal propagation of Chlamydia suis differed among isolates, and we identified cell lines outperforming those routinely used for Chlamydial isolation. We conclude that adaptation of the propagation procedure to the origin of the putative field isolate is highly recommended to improve the recovery rate.
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Assessment of Chlamydia suis Infection in Pig Farmers
Transboundary and emerging diseases, 2015Co-Authors: Leentje De Puysseleyr, Eric Cox, Kristien De Puysseleyr, Lutgart Braeckman, Servaas A Morré, Daisy VanrompayAbstract:Chlamydia suis infections are endemic in domestic pigs in Europe and can lead to conjunctivitis, pneumonia, enteritis and reproductive failure. Currently, the knowledge on the zoonotic potential of C. suis is limited. Moreover, the last decades, porcine tetracycline resistant C. suis strains have been isolated, which interfere with treatment of Chlamydial infections. In this study, the presence of C. suis was examined on nine Belgian pig farms, using Chlamydia culture and a C. suis specific real-time PCR in both pigs and farmers. In addition to diagnosis for C. suis, the farmers' samples were examined using a Chlamydia trachomatis PCR. Additionally, the Chlamydia isolates were tested for the presence of the tet(C) resistance gene. C. DNA was demonstrated in pigs on all farms, and eight of nine farmers were positive in at least one anatomical site. None of the farmers tested positive for C. trachomatis. Chlamydia suis isolates were obtained from pigs of eight farms. Nine porcine C. suis isolates possessing a tet(C) gene were retrieved, originating from three farms. Moreover, C. suis isolates were identified in three human samples, including one pharyngeal and two rectal samples. These findings suggest further research on the zoonotic transfer of C. suis from pigs to humans is needed.
Nicole Borel - One of the best experts on this subject based on the ideXlab platform.
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Chlamydia suis is associated with intestinal NF-κB activation in experimentally infected gnotobiotic piglets.
Pathogens and disease, 2020Co-Authors: Helen Aumayer, Cory Ann Leonard, Theresa Pesch, Barbara Prähauser, Sabina Wunderlin, Franco Guscetti, Nicole BorelAbstract:Chlamydia suis intestinal infection of single-animal experimental groups of gnotobiotic newborn piglets was previously reported to cause severe, temporary small intestinal epithelium damage. We investigated archived intestinal samples for pro-inflammatory nuclear factor kappa B (NF-κB) activation, Interleukin (IL)-6 and IL-8 production and immune cell influx. Samples were collected 2, 4 and 7 days post-inoculation with C. suis strain S45/6 or mock inoculum (control). Increased nuclear localization of epithelial NF-κB, representative of activation, in the jejunum and ileum of C. suis-infected animals, compared to uninfected controls, began by 2 days post-infection (dpi) and persisted through 7 dpi. Infected animals showed increased production of IL-8, peaking at 2 dpi, compared to controls. Infection-mediated CD45-positive immune cell influx into the jejunal lamina propria peaked at 7 dpi, when epithelial damage was largely resolved. Activation of NF-κB appears to be a key early event in the innate response of the unprimed porcine immune system challenged with C. suis. This results in an acute phase, coinciding with the most severe clinical symptoms, diarrhea and weight loss. Immune cells recruited shortly after infection remain present in the lamina propria during the recovery phase, which is characterized by reduced Chlamydial shedding and restored intestinal epithelium integrity.
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Isolation of Tetracycline-Resistant Chlamydia suis from a Pig Herd Affected by Reproductive Disorders and Conjunctivitis.
Antibiotics (Basel Switzerland), 2020Co-Authors: Christine Unterweger, Nicole Borel, Martina Jelocnik, Lukas Schwarz, René Brunthaler, Aleksandra Inic-kanada, Hanna MartiAbstract:Due to various challenges in diagnosing chlamydiosis in pigs, antibiotic treatment is usually performed before any molecular or antibiotic susceptibility testing. This could increase the occurrence of tetracycline-resistant Chlamydia (C.) suis isolates in the affected pig population and potentiate the reoccurrence of clinical signs. Here, we present a case of an Austrian pig farm, where tetracycline resistant and sensitive C. suis isolates were isolated from four finishers with conjunctivitis. On herd-level, 10% of the finishers suffered from severe conjunctivitis and sows showed a high percentage of irregular return to estrus. Subsequent treatment of whole-herd using oxytetracycline led to a significant reduction of clinical signs. Retrospective antibiotic susceptibility testing revealed tetracycline resistance and decreased susceptibility to doxycycline in half of the ocular C. suis isolates, and all isolates were able to partially recover following a single-dose tetracycline treatment in vitro. These findings were later confirmed in vivo, when all former clinical signs recurred three months later. This case report raises awareness of tetracycline resistance in C. suis and emphasizes the importance of preventative selection of tetracycline resistant C. suis isolates.
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Evaluating the antibiotic susceptibility of Chlamydia – new approaches for in vitro assays
Frontiers in microbiology, 2018Co-Authors: Hanna Marti, Nicole Borel, Deborah Dean, Cory Ann LeonardAbstract:Pigs are the natural hosts of Chlamydia suis, the only Chlamydia species known to spontaneously acquire homotypic resistance conferred by a class C tetracycline resistance gene. Various susceptibility assays have existed for several years, but there is no widely accepted, standardized assay to determine Chlamydial antibiotic susceptibility. In this study, we developed new approaches to determine the in vitro susceptibility of Chlamydia to different antibiotics in view of existing protocols. Specifically, the minimal inhibitory concentration (MIC) is based on a consensus of both inclusion number reduction and alteration of inclusion size and morphology upon antibiotic exposure. In addition to these, we employed a recovery assay, allowing observation of the Chlamydial response to drug removal and subsequent recovery, as compared to both continued exposure and to the unexposed control. We propose a simple and fast screening method to detect tetracycline resistant C. suis strains within 2 to 3 days with minimal use of consumables. For proof of principle, we evaluated the susceptibility of three C. suis field strains and the reference strain S45/6 to tetracycline, sulfamethoxazole, and penicillin, antibiotics commonly used to prevent respiratory and gastrointestinal diseases on fattening pig farms. We found that tetracycline sensitive strains can easily be distinguished from resistant strains using the evaluation parameters proposed in this study. Moreover, we report that S45/6 is sensitive to sulfamethoxazole while all evaluated C. suis field strains showed some degree of sulfamethoxazole resistance. Finally, we confirm that Penicillin G induces the Chlamydial stress response in all evaluated C. suis strains.
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Image_2_Evaluating the Antibiotic Susceptibility of Chlamydia – New Approaches for in Vitro Assays.TIF
2018Co-Authors: Hanna Marti, Nicole Borel, Deborah Dean, Cory A. LeonardAbstract:Pigs are the natural hosts of Chlamydia suis, the only Chlamydia species known to spontaneously acquire homotypic resistance conferred by a class C tetracycline resistance gene. Various susceptibility assays have existed for several years, but there is no widely accepted, standardized assay to determine Chlamydial antibiotic susceptibility. In this study, we developed new approaches to determine the in vitro susceptibility of Chlamydia to different antibiotics in view of existing protocols. Specifically, the minimal inhibitory concentration (MIC) is based on a consensus of both inclusion number reduction and alteration of inclusion size and morphology upon antibiotic exposure. In addition to these, we employed a recovery assay, allowing observation of the Chlamydial response to drug removal and subsequent recovery, as compared to both continued exposure and to the unexposed control. We propose a simple and fast screening method to detect tetracycline resistant C. suis strains within 2 to 3 days with minimal use of consumables. For proof of principle, we evaluated the susceptibility of three C. suis field strains and the reference strain S45/6 to tetracycline, sulfamethoxazole, and penicillin, antibiotics commonly used to prevent respiratory and gastrointestinal diseases on fattening pig farms. We found that tetracycline sensitive strains can easily be distinguished from resistant strains using the evaluation parameters proposed in this study. Moreover, we report that S45/6 is sensitive to sulfamethoxazole while all evaluated C. suis field strains showed some degree of sulfamethoxazole resistance. Finally, we confirm that Penicillin G induces the Chlamydial stress response in all evaluated C. suis strains.
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Data_Sheet_1_Evaluating the Antibiotic Susceptibility of Chlamydia – New Approaches for in Vitro Assays.pdf
2018Co-Authors: Hanna Marti, Nicole Borel, Deborah Dean, Cory A. LeonardAbstract:Pigs are the natural hosts of Chlamydia suis, the only Chlamydia species known to spontaneously acquire homotypic resistance conferred by a class C tetracycline resistance gene. Various susceptibility assays have existed for several years, but there is no widely accepted, standardized assay to determine Chlamydial antibiotic susceptibility. In this study, we developed new approaches to determine the in vitro susceptibility of Chlamydia to different antibiotics in view of existing protocols. Specifically, the minimal inhibitory concentration (MIC) is based on a consensus of both inclusion number reduction and alteration of inclusion size and morphology upon antibiotic exposure. In addition to these, we employed a recovery assay, allowing observation of the Chlamydial response to drug removal and subsequent recovery, as compared to both continued exposure and to the unexposed control. We propose a simple and fast screening method to detect tetracycline resistant C. suis strains within 2 to 3 days with minimal use of consumables. For proof of principle, we evaluated the susceptibility of three C. suis field strains and the reference strain S45/6 to tetracycline, sulfamethoxazole, and penicillin, antibiotics commonly used to prevent respiratory and gastrointestinal diseases on fattening pig farms. We found that tetracycline sensitive strains can easily be distinguished from resistant strains using the evaluation parameters proposed in this study. Moreover, we report that S45/6 is sensitive to sulfamethoxazole while all evaluated C. suis field strains showed some degree of sulfamethoxazole resistance. Finally, we confirm that Penicillin G induces the Chlamydial stress response in all evaluated C. suis strains.
Antonietta Di Francesco - One of the best experts on this subject based on the ideXlab platform.
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PREVALENCE OF ChlamydiaCEAE AND TETRACYCLINE RESISTANCE GENES IN WILD BOARS OF CENTRAL EUROPE
'Wildlife Disease Association', 2020Co-Authors: Amira Wahdan, Antonietta Di Francesco, Hanna Marti, Lea Rohner, Luca Nicola Bacciarini, Chiara Menegatti, And Nicole BorelAbstract:Our aim was to investigate the occurrence and distribution of Chlamydia suis and other Chlamydiaceae in the wild boar (Sus scrofa) population of Switzerland and Northern Italy and the detection of tetracycline resistance genes by PCR. We collected a total of 471 conjunctival swabs (n\ubc292), rectal swabs (n\ubc147), and lung tissue samples (n\ubc32) belonging to 292 wild boars. The prevalence of Chlamydiaceae in the investigated wild boar populations was very low (1.4%, 4/292). We found C. suis in rectal or conjunctival swabs but not in lung samples. The low Chlamydial prevalence might be attributed to limited contacts between wild boars and outdoor domestic pigs due to strict biosecurity measures or limited numbers of rural pig herds. The tetA(C) gene fragment was detected in six samples, which were all negative for Chlamydiaceae, and was probably not of Chlamydial origin but more likely from other bacteria. The low tetracycline resistance rate in wild boar might be explained by the lack of selective pressure. However, transmission of resistance genes from domestic pigs to wild boar or selective pressure in the environment could lead to the development and spread of tetracyclineresistant C. suis strains in wild boars
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The Chlamydia suis Genome Exhibits High Levels of Diversity, Plasticity, and Mobile Antibiotic Resistance: Comparative Genomics of a Recent Livestock Cohort Shows Influence of Treatment Regimes.
Genome biology and evolution, 2017Co-Authors: Helena M. B. Seth-smith, Antonietta Di Francesco, Manuela Donati, Adam Polkinghorne, Sabrina Wanninger, Hanna Marti, Nathan L. Bachmann, Nicole BorelAbstract:Chlamydia suis is an endemic pig pathogen, belonging to a fascinating genus of obligate intracellular pathogens. Of particular interest, this is the only Chlamydial species to have naturally acquired genes encoding for tetracycline resistance. To date, the distribution and mobility of the Tet-island are not well understood. Our study focused on whole genome sequencing of 29 C. suis isolates from a recent porcine cohort within Switzerland, combined with data from USA tetracycline-resistant isolates. Our findings show that the genome of C. suis is very plastic, with unprecedented diversity, highly affected by recombination and plasmid exchange. A large diversity of isolates circulates within Europe, even within individual Swiss farms, suggesting that C. suis originated around Europe. New World isolates have more restricted diversity and appear to derive from European isolates, indicating that historical strain transfers to the United States have occurred. The architecture of the Tet-island is variable, but the tetA(C) gene is always intact, and recombination has been a major factor in its transmission within C. suis. Selective pressure from tetracycline use within pigs leads to a higher number of Tet-island carrying isolates, which appear to be lost in the absence of such pressure, whereas the loss or gain of the Tet-island from individual strains is not observed. The Tet-island appears to be a recent import into the genome of C. suis, with a possible American origin.
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Selective Pressure Promotes Tetracycline Resistance of Chlamydia suis in Fattening Pigs.
PloS one, 2016Co-Authors: Sabrina Wanninger, Antonietta Di Francesco, Manuela Donati, Michael Hässig, Karolin Hoffmann, Helena M. B. Seth-smith, Hanna Marti, Nicole BorelAbstract:In pigs, Chlamydia suis has been associated with respiratory disease, diarrhea and conjunctivitis, but there is a high rate of inapparent C. suis infection found in the gastrointestinal tract of pigs. Tetracycline resistance in C. suis has been described in the USA, Italy, Switzerland, Belgium, Cyprus and Israel. Tetracyclines are commonly used in pig production due to their broad-spectrum activity and relatively low cost. The aim of this study was to isolate clinical C. suis samples in cell culture and to evaluate their antibiotic susceptibility in vitro under consideration of antibiotic treatment on herd level. Swab samples (n = 158) identified as C. suis originating from 24 farms were further processed for isolation, which was successful in 71% of attempts with a significantly higher success rate from fecal swabs compared to conjunctival swabs. The farms were divided into three treatment groups: A) farms without antibiotic treatment, B) farms with prophylactic oral antibiotic treatment of the whole herd consisting of trimethoprime, sulfadimidin and sulfathiazole (TSS), or C) farms giving herd treatment with chlortetracycline with or without tylosin and sulfadimidin (CTS). 59 isolates and their corresponding clinical samples were selected and tested for the presence or absence of the tetracycline resistance class C gene [tet(C)] by conventional PCR and isolates were further investigated for their antibiotic susceptibility in vitro. The phenotype of the investigated isolates was either classified as tetracycline sensitive (Minimum inhibitory concentration [MIC] < 2 μg/ml), intermediate (2 μg/ml ≤ MIC < 4 μg/ml) or resistant (MIC ≥ 4 μg/ml). Results of groups and individual pigs were correlated with antibiotic treatment and time of sampling (beginning/end of the fattening period). We found clear evidence for selective pressure as absence of antibiotics led to isolation of only tetracycline sensitive or intermediate strains whereas tetracycline treatment resulted in a greater number of tetracycline resistant isolates.
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Tetracycline Susceptibility in Chlamydia suis Pig Isolates.
PloS one, 2016Co-Authors: Manuela Donati, Nicole Borel, Andrea Balboni, Karine Laroucau, Rachid Aaziz, Fabien Vorimore, Federico Morandi, Edoardo Vecchio Nepita, Antonietta Di FrancescoAbstract:The aims of the present study were to assess the prevalence of Chlamydia suis in an Italian pig herd, determine the tetracycline susceptibility of C. suis isolates, and evaluate tet(C) and tetR(C) gene expression. Conjunctival swabs from 20 pigs were tested for C. suis by real-time polymerase chain reaction, and 55% (11) were positive. C. suis was then isolated from 11 conjunctival swabs resampled from the same herd. All positive samples and isolates were positive for the tet(C) resistance gene. The in vitro susceptibility to tetracycline of the C. suis isolates showed MIC values ranging from 0.5 to 4 μg/mL. Tet(C) and tetR(C) transcripts were found in all the isolates, cultured both in the absence and presence of tetracycline. This contrasts with other Gram-negative bacteria in which both genes are repressed in the absence of the drug. Further investigation into tet gene regulation in C. suis is needed.
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In vitro MICs of tetracycline against 11 Chlamydia suis isolates.
2016Co-Authors: Manuela Donati, Nicole Borel, Andrea Balboni, Karine Laroucau, Rachid Aaziz, Fabien Vorimore, Federico Morandi, Edoardo Vecchio Nepita, Antonietta Di FrancescoAbstract:In vitro MICs of tetracycline against 11 Chlamydia suis isolates.
Manuela Donati - One of the best experts on this subject based on the ideXlab platform.
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The Chlamydia suis Genome Exhibits High Levels of Diversity, Plasticity, and Mobile Antibiotic Resistance: Comparative Genomics of a Recent Livestock Cohort Shows Influence of Treatment Regimes.
Genome biology and evolution, 2017Co-Authors: Helena M. B. Seth-smith, Antonietta Di Francesco, Manuela Donati, Adam Polkinghorne, Sabrina Wanninger, Hanna Marti, Nathan L. Bachmann, Nicole BorelAbstract:Chlamydia suis is an endemic pig pathogen, belonging to a fascinating genus of obligate intracellular pathogens. Of particular interest, this is the only Chlamydial species to have naturally acquired genes encoding for tetracycline resistance. To date, the distribution and mobility of the Tet-island are not well understood. Our study focused on whole genome sequencing of 29 C. suis isolates from a recent porcine cohort within Switzerland, combined with data from USA tetracycline-resistant isolates. Our findings show that the genome of C. suis is very plastic, with unprecedented diversity, highly affected by recombination and plasmid exchange. A large diversity of isolates circulates within Europe, even within individual Swiss farms, suggesting that C. suis originated around Europe. New World isolates have more restricted diversity and appear to derive from European isolates, indicating that historical strain transfers to the United States have occurred. The architecture of the Tet-island is variable, but the tetA(C) gene is always intact, and recombination has been a major factor in its transmission within C. suis. Selective pressure from tetracycline use within pigs leads to a higher number of Tet-island carrying isolates, which appear to be lost in the absence of such pressure, whereas the loss or gain of the Tet-island from individual strains is not observed. The Tet-island appears to be a recent import into the genome of C. suis, with a possible American origin.
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Selective Pressure Promotes Tetracycline Resistance of Chlamydia suis in Fattening Pigs.
PloS one, 2016Co-Authors: Sabrina Wanninger, Antonietta Di Francesco, Manuela Donati, Michael Hässig, Karolin Hoffmann, Helena M. B. Seth-smith, Hanna Marti, Nicole BorelAbstract:In pigs, Chlamydia suis has been associated with respiratory disease, diarrhea and conjunctivitis, but there is a high rate of inapparent C. suis infection found in the gastrointestinal tract of pigs. Tetracycline resistance in C. suis has been described in the USA, Italy, Switzerland, Belgium, Cyprus and Israel. Tetracyclines are commonly used in pig production due to their broad-spectrum activity and relatively low cost. The aim of this study was to isolate clinical C. suis samples in cell culture and to evaluate their antibiotic susceptibility in vitro under consideration of antibiotic treatment on herd level. Swab samples (n = 158) identified as C. suis originating from 24 farms were further processed for isolation, which was successful in 71% of attempts with a significantly higher success rate from fecal swabs compared to conjunctival swabs. The farms were divided into three treatment groups: A) farms without antibiotic treatment, B) farms with prophylactic oral antibiotic treatment of the whole herd consisting of trimethoprime, sulfadimidin and sulfathiazole (TSS), or C) farms giving herd treatment with chlortetracycline with or without tylosin and sulfadimidin (CTS). 59 isolates and their corresponding clinical samples were selected and tested for the presence or absence of the tetracycline resistance class C gene [tet(C)] by conventional PCR and isolates were further investigated for their antibiotic susceptibility in vitro. The phenotype of the investigated isolates was either classified as tetracycline sensitive (Minimum inhibitory concentration [MIC] < 2 μg/ml), intermediate (2 μg/ml ≤ MIC < 4 μg/ml) or resistant (MIC ≥ 4 μg/ml). Results of groups and individual pigs were correlated with antibiotic treatment and time of sampling (beginning/end of the fattening period). We found clear evidence for selective pressure as absence of antibiotics led to isolation of only tetracycline sensitive or intermediate strains whereas tetracycline treatment resulted in a greater number of tetracycline resistant isolates.
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Tetracycline Susceptibility in Chlamydia suis Pig Isolates.
PloS one, 2016Co-Authors: Manuela Donati, Nicole Borel, Andrea Balboni, Karine Laroucau, Rachid Aaziz, Fabien Vorimore, Federico Morandi, Edoardo Vecchio Nepita, Antonietta Di FrancescoAbstract:The aims of the present study were to assess the prevalence of Chlamydia suis in an Italian pig herd, determine the tetracycline susceptibility of C. suis isolates, and evaluate tet(C) and tetR(C) gene expression. Conjunctival swabs from 20 pigs were tested for C. suis by real-time polymerase chain reaction, and 55% (11) were positive. C. suis was then isolated from 11 conjunctival swabs resampled from the same herd. All positive samples and isolates were positive for the tet(C) resistance gene. The in vitro susceptibility to tetracycline of the C. suis isolates showed MIC values ranging from 0.5 to 4 μg/mL. Tet(C) and tetR(C) transcripts were found in all the isolates, cultured both in the absence and presence of tetracycline. This contrasts with other Gram-negative bacteria in which both genes are repressed in the absence of the drug. Further investigation into tet gene regulation in C. suis is needed.
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In vitro MICs of tetracycline against 11 Chlamydia suis isolates.
2016Co-Authors: Manuela Donati, Nicole Borel, Andrea Balboni, Karine Laroucau, Rachid Aaziz, Fabien Vorimore, Federico Morandi, Edoardo Vecchio Nepita, Antonietta Di FrancescoAbstract:In vitro MICs of tetracycline against 11 Chlamydia suis isolates.
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Prevalence of Chlamydial Infections in Fattening Pigs and Their Influencing Factors
PloS one, 2015Co-Authors: Karolin Hoffmann, Antonietta Di Francesco, Manuela Donati, Sabrina Wanninger, Michael Hässig, Franziska Schott, Xaver Sidler, Nicole BorelAbstract:Chlamydial infections in pigs are associated with respiratory disease, diarrhea, conjunctivitis and other pathologies. The aim of this study was to define the prevalence of Chlamydiaceae in Swiss fattening pigs by applying sensitive and specific detection methods and to correlate prior antibiotic treatment and farm related factors with differences in prevalence. Conjunctival and fecal swabs were collected from 636 pigs in 29 Swiss fattening pig farms with and without antibiotic treatment, at the beginning and the end of the fattening period. The swabs were screened by real-time PCR for Chlamydiaceae. For the Chlamydial detection and species-identification, a DNA-microarray analysis was performed. All farms were positive for Chlamydiaceae with 94.3 and 92.0% prevalence in fecal swabs as well as 45.9 and 32.6% in conjunctival swabs at the first and second time points, respectively. Antibiotic treatment could not clear the infection on herd level. Potential contact with wild boars was a significant risk factor, while hygiene criteria did not influence Chlamydial prevalence. A correlation of Chlamydial positivity to diarrhea, but not to conjunctivitis was evident. Chlamydia suis was the predominant species. Mixed infections with C. suis and C. pecorum were common, with a substantial increase in C. pecorum positivity at the end of the fattening period, and this finding was associated with ruminant contact. C. abortus was detected in one conjunctival swab. In this study, C. suis inhabited the intestinal tract of nearly all examined pigs, implying a long-term infection. C. pecorum was also common and might be transmitted to pigs by ruminants.
Eric Cox - One of the best experts on this subject based on the ideXlab platform.
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Assessment of Chlamydia suis Infection in Pig Farmers
Transboundary and emerging diseases, 2015Co-Authors: Leentje De Puysseleyr, Eric Cox, Kristien De Puysseleyr, Lutgart Braeckman, Servaas A Morré, Daisy VanrompayAbstract:Chlamydia suis infections are endemic in domestic pigs in Europe and can lead to conjunctivitis, pneumonia, enteritis and reproductive failure. Currently, the knowledge on the zoonotic potential of C. suis is limited. Moreover, the last decades, porcine tetracycline resistant C. suis strains have been isolated, which interfere with treatment of Chlamydial infections. In this study, the presence of C. suis was examined on nine Belgian pig farms, using Chlamydia culture and a C. suis specific real-time PCR in both pigs and farmers. In addition to diagnosis for C. suis, the farmers' samples were examined using a Chlamydia trachomatis PCR. Additionally, the Chlamydia isolates were tested for the presence of the tet(C) resistance gene. C. DNA was demonstrated in pigs on all farms, and eight of nine farmers were positive in at least one anatomical site. None of the farmers tested positive for C. trachomatis. Chlamydia suis isolates were obtained from pigs of eight farms. Nine porcine C. suis isolates possessing a tet(C) gene were retrieved, originating from three farms. Moreover, C. suis isolates were identified in three human samples, including one pharyngeal and two rectal samples. These findings suggest further research on the zoonotic transfer of C. suis from pigs to humans is needed.
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The immune response against Chlamydia suis genital tract infection partially protects against re-infection
Veterinary Research, 2014Co-Authors: Evelien De Clercq, Bert Devriendt, Lizi Yin, Koen Chiers, Eric Cox, Daisy VanrompayAbstract:The aim of the present study was to reveal the characteristic features of genital Chlamydia suis infection and re-infection in female pigs by studying the immune response, pathological changes, replication of Chlamydial bacteria in the genital tract and excretion of viable bacteria. Pigs were intravaginally infected and re-infected with C. suis strain S45, the type strain of this species. We demonstrated that S45 is pathogenic for the female urogenital tract. Chlamydia replication occurred throughout the urogenital tract, causing inflammation and pathology. Furthermore, genital infection elicited both cellular and humoral immune responses. Compared to the primo-infection of pigs with C. suis, re-infection was characterized by less severe macroscopic lesions and less Chlamydial elementary bodies and inclusions in the urogenital tract. This indicates the development of a certain level of protection following the initial infection. Protective immunity against re-infection coincided with higher Chlamydia-specific IgG and IgA antibody titers in sera and vaginal secretions, higher proliferative responses of peripheral blood mononuclear cells (PBMC), higher percentages of blood B lymphocytes, monocytes and CD8+ T cells and upregulated production of IFN-γ and IL-10 by PBMC.
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Evaluation of the presence and zoonotic transmission of Chlamydia suis in a pig slaughterhouse
BMC Infectious Diseases, 2014Co-Authors: Kristien De Puysseleyr, Eric Cox, Leentje De Puysseleyr, Hendrik Dhondt, Tom Geens, Lutgart Braeckman, Servaas A Morré, Daisy VanrompayAbstract:Background A significant number of studies on pig farms and wild boars worldwide, demonstrate the endemic presence of Chlamydia suis in pigs. However, the zoonotic potential of this pathogen, phylogenetically closely related to Chlamydia trachomatis , is still uninvestigated. Therefore, this study aims to examine the zoonotic transmission in a Belgian pig abattoir. Methods Presence of Chlamydia suis in pigs, contact surfaces, air and employees was assessed using a Chlamydia suis specific real-time PCR and culture. Furthermore, Chlamydia suis isolates were tested for the presence of the tet(C) gene. Results Chlamydia suis bacteria could be demonstrated in samples from pigs, the air and contact surfaces. Moreover, eye swabs of two employees were positive for Chlamydia suis by both PCR and culture. The tet (C) gene was absent in both human Chlamydia suis isolates and no clinical signs were reported. Conclusions These findings suggest the need for further epidemiological and clinical research to elucidate the significance of human ocular Chlamydia suis infections.
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Evaluation of the presence and zoonotic transmission of Chlamydia suis in a pig slaughterhouse.
BMC infectious diseases, 2014Co-Authors: Kristien De Puysseleyr, Eric Cox, Leentje De Puysseleyr, Hendrik Dhondt, Tom Geens, Lutgart Braeckman, Servaas A Morré, Daisy VanrompayAbstract:Background: A significant number of studies on pig farms and wild boars worldwide, demonstrate the endemic presence of Chlamydia suis in pigs. However, the zoonotic potential of this pathogen, phylogenetically closely related to Chlamydia trachomatis, is still uninvestigated. Therefore, this study aims to examine the zoonotic transmission in a Belgian pig abattoir. Methods: Presence of Chlamydia suis in pigs, contact surfaces, air and employees was assessed using a Chlamydia suis specific real-time PCR and culture. Furthermore, Chlamydia suis isolates were tested for the presence of the tet(C) gene. Results: Chlamydia suis bacteria could be demonstrated in samples from pigs, the air and contact surfaces. Moreover, eye swabs of two employees were positive for Chlamydia suis by both PCR and culture. The tet(C) gene was absent in both human Chlamydia suis isolates and no clinical signs were reported.
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Development and validation of a real-time PCR for Chlamydia suis diagnosis in swine and humans.
PloS one, 2014Co-Authors: Kristien De Puysseleyr, Eric Cox, Leentje De Puysseleyr, Julie Geldhof, Daisy VanrompayAbstract:Pigs are the natural host for Chlamydia suis, a pathogen which is phylogenetically highly related to the human pathogen C. trachomatis. Chlamydia suis infections are generally treated with tetracyclines. In 1998, tetracyline resistant C. suis strains emerged on U.S. pig farms and they are currently present in the Belgian, Cypriote, German, Israeli, Italian and Swiss pig industry. Infections with tetracycline resistant C. suis strains are mainly associated with severe reproductive failure leading to marked economical loss. We developed a sensitive and specific TaqMan probe-based C. suis real-time PCR for examining clinical samples of both pigs and humans. The analytical sensitivity of the real-time PCR is 10 rDNA copies/reaction without cross-amplifying DNA of other Chlamydia species. The PCR was successfully validated using conjunctival, pharyngeal and stool samples of slaughterhouse employees, as well as porcine samples from two farms with evidence of reproductive failure and one farm without clinical disease. Chlamydia suis was only detected in diseased pigs and in the eyes of humans. Positive humans had no clinical complaints. PCR results were confirmed by culture in McCoy cells. In addition, Chlamydia suis isolates were also examined by the tet(C) PCR, designed for demonstrating the tetracycline resistance gene tet(C). The tet(C) gene was only present in porcine C. suis isolates.