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2022
Fawzy, A., A. - S. Giel, L. Fenske, A. Bach, C. Herden, K. Engel, E. Heuser, M. Boelhauve, R. G. Ulrich, K. Vogel, et al., "Development and validation of a triplex real-time qPCR for sensitive detection and quantification of major rat bite fever pathogen Streptobacillus moniliformis.", Journal of microbiological methods, vol. 199, pp. 106525, 2022. Abstract

Streptobacillus (S.) moniliformis is the most important pathogen causing rat bite fever (RBF) worldwide. This zoonotic pathogen is understudied mainly due to difficulties in culturing S. moniliformis as a fastidious microorganism. Therefore, advances in molecular detection techniques are highly needed, especially with regard to the widespread availability of real-time quantitative (q) PCR in laboratories. In this study, we aimed to develop a qPCR for the identification of Streptobacillus species and quantification of S. moniliformis in clinical samples, especially those derived from tissue samples of animal origin. We optimized a previously described PCR protocol in order to develop a qPCR, which can detect different Streptobacillus species with high specificity and is simultaneously able to quantitate S. moniliformis in different clinical matrices. The qPCR exhibited a limit of detection (LOD) of 21 copies/reaction representing ~4-5 streptobacilli, while the limit of quantification (LOQ) was 2.1 × 10 copies/reaction. It was also more sensitive than conventional PCR by two orders of magnitude and proved to have a substantial agreement (Kappa 0.74) compared to it with a superior detection rate in 374 samples from wild rats, laboratory rats and animals from holdings of wild-trapped rats. To conclude, the qPCR described in this study is an important molecular tool that is able to quantify S. moniliformis in tissue samples of animal origin. It represents a suitable tool for future establishment and evaluation of other molecular assays that are highly needed for a better understanding of epidemiology and pathophysiology of RBF. In experimental studies, it will also be useful for titration purposes since the quantification of the organism using classical plate counting technique is problematic and inaccurate.

Fenske, L., I. Noll, J. Blom, C. Ewers, T. Semmler, A. Fawzy, and T. Eisenberg, "A dominant clonal lineage of Streptococcus uberis in cattle in Germany.", Antonie van Leeuwenhoek, vol. 115, issue 7, pp. 857-870, 2022. Abstract

Bovine mastitis causes enormous economic losses in the dairy industry with Streptococcus uberis as one of the most common bacterial pathogens causing clinical and subclinical variations. In most cases mastitis can be cured by intramammary administration of antimicrobial agents. However, the severity of the clinical manifestations can vary greatly from mild to severe symtoms. In this study, a comparative genomic analysis of 24 S. uberis isolates from three dairy farms in Germany, affected by different courses of infection was conducted. While there were sporadic mild infections in farm A and B, a large number of infections were observed within a very short period of time in farm C. The comparison of virulence genes, antimicrobial resistance genes and prophage regions revealed no features that might be responsible for this severe course. However, almost all isolates from farm C showed the same, novel MLST profile (ST1373), thus a clonal outbreak cannot be excluded, whereby the actual reason for the particular virulence remains unknown. This study demonstrates the importance of extensive metagenomic studies, including the host genomes and the environment, to gain further evidence on the pathogenicity of S. uberis.

Eisenberg, T., K. Mühldorfer, M. Erhard, A. Fawzy, S. Kehm, C. Ewers, T. Semmler, J. Blom, A. Lipski, J. Rau, et al., "sp. nov., sp. nov. and sp. nov., three novel species of the genus, and emended description of .", International journal of systematic and evolutionary microbiology, vol. 72, issue 7, 2022. Abstract

Seven genotypically distinct strains assigned to the genus were isolated in different laboratories from several animal sources. Strain D17_0559-3-2-1 and three further strains were isolated from samples of duck, pig and goose. The strains had >99 % 16S rRNA gene sequence similarity to each other and to strain VA92-K48 and two further strains isolated from samples of medical leech and a turtle. The closest related type strains to the seven strains were those of (96.74 %) and (95.93 %). Average nucleotide identity, amino acid identity and DNA-DNA hybridization results showed that the strains represented two separate novel species. One further phylogenetically distinct strain (165301687) was isolated from fox urine. The strain had highest 16S rRNA gene sequence similarity to the type strains of (95.67 %), followed by (95.58 %) and (94.22 %) and represented a further novel species. Chemotaxonomic and physiological data of the novel strains were assessed, but failed to unequivocally differentiate the novel species from existing members of the genus. MALDI-TOF MS data proved the discrimination of at least strain 165301687 from all currently described species. Based on the presented phylogenomic and physiological data, we propose three novel species, sp. nov. with strain D17_0559-3-2-1 (=DSM 111258= CIP 111884=CCM 9044) as type strain, sp. nov. with strain VA92-K48 (=DSM 106012=LMG 30351=CIP 111492) as type strain and sp. nov. with strain 165301687 (=DSM 106013= LMG 30352= CIP 111494) as type strain.

2021
Matt, U., J. Schmiedel, A. Fawzy, J. Trauth, K. Schmidt, K. Vogel, S. Herold, T. Karrasch, C. Imirzalioglu, and T. Eisenberg, "Infection in a Young Immunocompetent Male Caused by Streptobacillus felis, a Putative Zoonotic Microorganism Transmitted by Cats.", Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, vol. 72, issue 10, pp. 1826-1829, 2021. Abstract

Rat bite fever (RBF) is predominantly caused by Streptobacillus moniliformis. We report a human infection with Streptobacillus felis. Clinical presentation was consistent with RBF, but serologic testing was negative for S moniliformis. Eventually, S felis-specific sequences were detected in skin lesions of the patient and in the oropharynx of local cats.

Eisenberg, T., S. Gronow, J. Falgenhauer4, C. Imirzalioglu, K. Mühldorfer, J. Rau, J. Blom, A. Fawzy, S. P. Glaeser, and P. Kämpfer, "Sneathia vaginalis sp. nov. (Fusobacteriales, Leptotrichiaceae) as a replacement of the species ‘Sneathia amnii’ Harwich et al. 2012 and ‘Leptotrichia amnionii’ Shukla et al. 2002, and emended description of Sneathia Collins et al. 2001", International Journal of systemic and evolutionary microbiology, vol. 71, issue 3, 2021.
2020
Eisenberg, T., A. Fawzy, U. Kaim, A. Nesseler, K. Riße, I. Völker, S. Hechinger, N. Schauerte, C. Geiger, T. Knauf-Witzens, et al., "Chronic wasting associated with Chlamydia pneumoniae in three ex situ breeding facilities for tropical frogs", Antonie van Leeuwenhoek, vol. 113, issue 12, pp. 2139 - 2154, 2020. Abstract

A number of different Chlamydia spp. have been detected in the class Amphibia with C. pneumoniae being the predominant species involved. Chlamydiae have been linked to mass mortality events, thereby representing significant pathogens that deserve attention with respect to worldwide amphibian decline. We here present six cases of chlamydiosis and asymptomatic chlamydial infections in different frog species from three ex situ amphibian conservation facilities. Clinical signs predominantly characterised by regurgitation, chronic wasting, lethargy and suspended breeding were associated with C. pneumoniae infection. Despite various treatment regimens, it was not possible to clear infections. However, intra vitam diagnostics succeeded from skin, faeces and urine for the first time.

Fawzy, A., M. Zschöck, C. Ewers, and T. Eisenberg, "Development of a hierarchical typing approach for Mycobacterium avium subsp. paratuberculosis (MAP) and characterization of MAP field cultures from Central Germany", Journal of applied microbiology, vol. 129, issue 5, pp. 1193 - 1206, 2020. Abstract

AIMSDevelopment of a novel hierarchical Mycobacterium avium subsp. paratuberculosis (MAP) typing approach and characterization of MAP field cultures in Central Germany.

METHODS AND RESULTS

By combining single nucleotide polymorphisms (SNPs) and mycobacterial interspersed repetitive unit-variable number tandem repeat, we developed a highly discriminating and phylogenetically accurate hierarchical MAP typing approach. Moreover, a novel stepwise workflow was employed to reduce the number of SNP reactions required making the typing approach more affordable. MAP field cultures (n = 142) from dairy herds in Central Germany were classified as cattle type and showed a high level of heterogeneity. Intra-herd multiple genotypes were evident in (13-25%) of the investigated herds.

CONCLUSIONS

The hierarchical MAP typing approach proved to be useful in fine discrimination between MAP cultures within limited geographical regions. This could potentially be used in unravelling MAP transmission chains in the respective regions. The observed heterogeneity in some herds is assumed to be due to either multiple introductions through inter-herd trade or intra-herd evolution over time.

SIGNIFICANCE AND IMPACT OF THE STUDY

Future MAP epidemiological studies will benefit from the advantages of the novel hierarchical typing approach. The SNP number reduction approach employed here could be extrapolated for other analogous pathogens.

Eisenberg, T., K. Schlez, A. Fawzy, I. Völker, S. Hechinger, M. Curić, N. Schauerte, C. Geiger, J. Blom, and H. C. Scholz, "Expanding the host range: infection of a reptilian host (Furcifer pardalis) by an atypical Brucella strain", Antonie van Leeuwenhoek, vol. 113, issue 10, pp. 1531 - 1537, 2020. Abstract

Atypical brucellae show deviant phenotypes and/or genotypes. Besides Brucella inopinata, B. microti and B. vulpis, atypical strains have been described infecting humans, rodents, amphibians and fish. They represent potential zoonotic agents. Here, we provide evidence that reptiles as the remaining poikilothermic vertebrate class also represent susceptible hosts for atypical Brucella.

Eisenberg, T., C. Heydel, E. Prenger-Berninghoff, A. Fawzy, U. Kling, V. Akimkin, T. Semmler, K. Mühldorfer, P. Kämpfer, J. Blom, et al., "Streptobacillus canis sp. nov. isolated from a dog", International journal of systematic and evolutionary microbiology, vol. 70, issue 4, pp. 2648 - 2656, 2020. Abstract

From a phlegmon in a dog an aerobic and facultatively anaerobic, indole-, oxidase- and catalase-negative, non-motile bacterium was isolated in 2019 in Germany that stained Gram-negative and showed a pleomorphic, rod-shaped, non-spore-forming appearance. Based on the results of 16S rRNA gene sequence analyses, strain IHIT1603-19T was assigned to the genus Streptobacillus with sequence similarities of 98.6, 98.0, 97.9, 97.1 and 94.4 % to the type strains of Streptobacillus felis, Streptobacillus notomytis, Streptobacillus ratti, Streptobacillus moniliformis and Streptobacillus hongkongensis, respectively. Strain IHIT1603-19T could also clearly be differentiated from other Streptobacillus species by rpoB, groEL and recA gene, nucleotide and amino acid sequence analyses as well as by core genome phylogeny. Regarding DNA-DNA relatedness, strain IHIT1603-19T demonstrated an average nucleotide identity of 83.00 and 82.28 % compared to S. felis 131000547T and S. moniliformis DSM 12112T, respectively. Chemotaxonomic and physiological data of strain IHIT1603-19T were in congruence with other closely related members of the family Leptotrichiaceae, represented by highly similar enzyme profiles and fatty acid patterns. MALDI-TOF MS analysis also proved suitable in unequivocally discriminating strain IHIT1603-19T from all currently described taxa of the genus Streptobacillus. On the basis of these data, we propose the novel species Streptobacillus canis sp. nov. with the type strain IHIT1603-19T (=DSM 110501T=CCUG 74118T=CIP 111795T). The G+C content of the DNA of the type strain is 26.6 mol%, genome size is 1.60 Mbp.

Fawzy, A., J. Rau, K. Riße, N. Schauerte, C. Geiger, J. Blom, C. Imirzalioglu, J. Falgenhauer, A. Bach, C. Herden, et al., "Streptobacillus felis, a member of the oropharynx microbiota of the Felidae, isolated from a tropical rusty-spotted cat", Antonie van Leeuwenhoek, vol. 113, issue 10, pp. 1455 - 1465, 2020. Abstract

Streptobacillus felis is a fastidious microorganism and a novel member of the potentially zoonotic bacteria causing rat bite fever. Since its description, this is the second isolation of S. felis in a diseased member of the Felidae. Interestingly, the strain from this study was isolated from a zoo held, rusty-spotted cat (Prionailurus rubiginosus), with pneumonia, thereby indicating a possible broader host range in feline species. A recent preliminary sampling of domestic cats (Felis silvestris forma catus) revealed that this microorganism is common in the oropharynx, suggesting that S. felis is a member of their normal microbiota. Due to unawareness, fastidiousness, antibiotic sensitivity and lack of diagnostics the role of S. felis as a cat and human pathogen might be under-reported as with other Streptobacillus infections. More studies are necessary to elucidate the role of S. felis in domestic cats and other Felidae in order to better estimate its zoonotic potential.

2019
Eisenberg, T., S. Gronow, J. Falgenhauer, C. Imirzalioglu, K. Mühldorfer, J. Rau, J. Blom, A. Fawzy, S. P. Glaeser, and P. Kämpfer, "sp. nov. () as a replacement of the species '' Harwich . 2012 and '' Shukla . 2002, and emended description of Collins . 2001.", International journal of systematic and evolutionary microbiology, vol. 71, issue 3, 2019. Abstract

Members of the genus are fastidious bacteria that predominantly colonise the female genital tract and are significantly associated with reproductive disorders and genital and neonatal disease. From a taxonomical perspective, the genus only comprises the species . Numerous reports on a second species, '', have been published, but the name has never been validated. The same is the case for '', which was previously shown to belong to the same species as ''. We studied strains DSM 16631 and DSM 16630, which have been identified and deposited as '' previously. At the time of isolation, these strains were found to be most closely related to, but clearly different from, based on 16S rRNA gene sequence similarities. Both strains proved to be almost indistinguishable from '' based on molecular, morphological and physiological traits. The 16S rRNA gene sequence analysis revealed that strain DSM 16631 was assigned to the genus with a sequence similarity of 95.47 % to CCUG 41628, followed by type strains of (93.03 %), (92.68 %) and (91.97 %) as the next closely related members of the . The novel species was also clearly differentiated from other related taxa by core genome phylogeny, average nucleotide and amino acid identities, DNA-DNA hybridization and MALDI-TOF MS. With respect to chemotaxonomic and physiological patterns, strains DSM 16631 and DSM 16630 were again highly similar to . On the basis of these data, we propose the novel species sp. nov. with the type strain DSM 16631 (=CCUG 52977=CCUG 52889A) and a second strain DSM 16630 (=CCUG 52976=CCUG 52888) that were both isolated from bloodstream infections in women with puerperal fever in France. The G+C content of the DNA of the type strain is 28.4 mol% and the genome size is 1.28 Mbp. Based on the observed extremely high similarities of genotypic and phenotypic traits of the novel proposed species to those reported for '', we recommend using this new name in all further publications on this taxon.

Eisenberg, T., S. Gronow, J. Falgenhauer, C. Imirzalioglu, K. Mühldorfer, J. Rau, J. Blom, A. Fawzy, S. P. Glaeser, and P. Kämpfer, "sp. nov. () as a replacement of the species '' Harwich . 2012 and '' Shukla . 2002, and emended description of Collins . 2001.", International journal of systematic and evolutionary microbiology, vol. 71, issue 3, 2019. Abstract

Members of the genus are fastidious bacteria that predominantly colonise the female genital tract and are significantly associated with reproductive disorders and genital and neonatal disease. From a taxonomical perspective, the genus only comprises the species . Numerous reports on a second species, '', have been published, but the name has never been validated. The same is the case for '', which was previously shown to belong to the same species as ''. We studied strains DSM 16631 and DSM 16630, which have been identified and deposited as '' previously. At the time of isolation, these strains were found to be most closely related to, but clearly different from, based on 16S rRNA gene sequence similarities. Both strains proved to be almost indistinguishable from '' based on molecular, morphological and physiological traits. The 16S rRNA gene sequence analysis revealed that strain DSM 16631 was assigned to the genus with a sequence similarity of 95.47 % to CCUG 41628, followed by type strains of (93.03 %), (92.68 %) and (91.97 %) as the next closely related members of the . The novel species was also clearly differentiated from other related taxa by core genome phylogeny, average nucleotide and amino acid identities, DNA-DNA hybridization and MALDI-TOF MS. With respect to chemotaxonomic and physiological patterns, strains DSM 16631 and DSM 16630 were again highly similar to . On the basis of these data, we propose the novel species sp. nov. with the type strain DSM 16631 (=CCUG 52977=CCUG 52889A) and a second strain DSM 16630 (=CCUG 52976=CCUG 52888) that were both isolated from bloodstream infections in women with puerperal fever in France. The G+C content of the DNA of the type strain is 28.4 mol% and the genome size is 1.28 Mbp. Based on the observed extremely high similarities of genotypic and phenotypic traits of the novel proposed species to those reported for '', we recommend using this new name in all further publications on this taxon.

Eisenberg, T., S. Gronow, J. Falgenhauer, C. Imirzalioglu, K. Mühldorfer, J. Rau, J. Blom, A. Fawzy, S. P. Glaeser, and P. Kämpfer, "sp. nov. () as a replacement of the species '' Harwich . 2012 and '' Shukla . 2002, and emended description of Collins . 2001.", International journal of systematic and evolutionary microbiology, vol. 71, issue 3, 2019. Abstract

Members of the genus are fastidious bacteria that predominantly colonise the female genital tract and are significantly associated with reproductive disorders and genital and neonatal disease. From a taxonomical perspective, the genus only comprises the species . Numerous reports on a second species, '', have been published, but the name has never been validated. The same is the case for '', which was previously shown to belong to the same species as ''. We studied strains DSM 16631 and DSM 16630, which have been identified and deposited as '' previously. At the time of isolation, these strains were found to be most closely related to, but clearly different from, based on 16S rRNA gene sequence similarities. Both strains proved to be almost indistinguishable from '' based on molecular, morphological and physiological traits. The 16S rRNA gene sequence analysis revealed that strain DSM 16631 was assigned to the genus with a sequence similarity of 95.47 % to CCUG 41628, followed by type strains of (93.03 %), (92.68 %) and (91.97 %) as the next closely related members of the . The novel species was also clearly differentiated from other related taxa by core genome phylogeny, average nucleotide and amino acid identities, DNA-DNA hybridization and MALDI-TOF MS. With respect to chemotaxonomic and physiological patterns, strains DSM 16631 and DSM 16630 were again highly similar to . On the basis of these data, we propose the novel species sp. nov. with the type strain DSM 16631 (=CCUG 52977=CCUG 52889A) and a second strain DSM 16630 (=CCUG 52976=CCUG 52888) that were both isolated from bloodstream infections in women with puerperal fever in France. The G+C content of the DNA of the type strain is 28.4 mol% and the genome size is 1.28 Mbp. Based on the observed extremely high similarities of genotypic and phenotypic traits of the novel proposed species to those reported for '', we recommend using this new name in all further publications on this taxon.

Mühldorfer, K., J. Rau, A. Fawzy, C. Heydel, S. P. Glaeser, M. van der Linden, P. Kutzer, T. Knauf-Witzens, M. Hanczaruk, A. S. Eckert, et al., "Streptococcus castoreus, an uncommon group A Streptococcus in beavers.", Antonie van Leeuwenhoek, vol. 112, issue 11, pp. 1663-1673, 2019. Abstract

Streptococcus castoreus is a rarely encountered beta-haemolytic group A Streptococcus with high tropism for the beaver as host. Based on 27 field isolates under study, evidence strongly suggests that S. castoreus behaves as an opportunistic pathogen in beavers. Although it belongs to the resident mucosal microbiota, this Streptococcus species is associated with purulent lesions in diseased animals. With few exceptions, isolates proved to be highly similar in a panel of phenotypic (including biochemistry, resistance pattern, MALDI-TOF mass spectrometry and Fourier transform-infrared spectroscopy) and classic molecular (16S rRNA and sodA gene) analyses, and thus did not show any specific pattern according to host species or spatio-temporal origin. Conversely, S. castoreus isolates were differentiated into a multitude of pulsed-field gel electrophoresis 'pulsotypes' that did not seem to reflect true epidemiologic lineages. In contrast, single reactions of genomic fingerprinting using BOX-, (GTG)- and RAPD-PCRs revealed at least subclusters with respect to host species, geographic origin or year, and confirmed the co-colonization of individuals with more than one isolate. In addition to isolates from free-ranging Eurasian beavers (Castor fiber), this study includes S. castoreus from captive North American beavers (Castor canadensis) for the first time.

2018
Michel, V., C. Ulber, D. Pöhle, B. Köpke, K. Engel, U. Kaim, A. Fawzy, S. Funk, J. Fornefett, C. G. Baums, et al., "Clinical infection in house rats (Rattus rattus) caused by Streptobacillus notomytis.", Antonie van Leeuwenhoek, vol. 111, issue 10, pp. 1955-1966, 2018 Oct. Abstract

Rat bite fever is an under-reported, under-diagnosed emerging zoonosis with worldwide distribution. Besides Spirillum minus, Streptobacillus moniliformis is the major causative microorganism although it usually colonises rats without any clinical signs. A group of house rats (Rattus rattus) kept in a zoo exhibition for educational purposes suffered from neurological signs including disorientation, torticollis, stall walking, ataxia and death. Gross pathological and histo-pathological examinations of the investigated rats revealed high-grade otitis interna et media, from which Streptobacillus notomytis was isolated in pure culture or as the predominant microorganism. This case series underlines a previously expressed hypothesis that R. rattus might be naturally colonised with S. notomytis, whereas the traditional rat bite fever organism, S. moniliformis, might be restricted to the Norway rat (Rattus norvegicus). However, the general paucity of Streptobacillus isolates, especially from their respective animal hosts, precludes definitive proof of these host tropisms. This is the first report of S. notomytis detection outside Asia and Australia and the first evidence for its role as a facultative pathogen in house rats.

Fawzy, A., M. Zschoeck, C. Ewers, and T. Eisneberg, "Genotyping methods and molecular epidemiology of Mycobacterium avium subsp. paratuberculosis (MAP)", Int J Vet Sci Med, vol. 6, issue 2, pp. 258-264, 2018. map_genotyping.pdf
2017
Eisenberg, T., J. Rau, U. Westerhüs, T. Knauf-Witzens, A. Fawzy, K. Schlez, M. Zschöck, E. Prenger-Berninghoff, C. Heydel, R. Sting, et al., "Streptococcus agalactiae in elephants - A comparative study with isolates from human and zoo animal and livestock origin.", Veterinary microbiology, vol. 204, pp. 141-150, 2017 May. Abstract

Streptococcus (S.) agalactiae represents a significant pathogen for humans and animals. However, there are only a few elderly reports on S. agalactiae infections in wild and zoo elephants even though this pathogen has been isolated comparatively frequently in these endangered animal species. Consequently, between 2004 and 2015, we collected S. agalactiae isolates from African and Asian elephants (n=23) living in four different zoos in Germany. These isolates were characterised and compared with isolates from other animal species (n=20 isolates) and humans (n=3). We found that the isolates from elephants can be readily identified by classical biochemistry and MALDI-TOF mass spectrometry. Further characterisations for epidemiological issues were achieved using Fourier transform-infrared spectroscopy, capsule typing and molecular fingerprinting (PFGE, RAPD PCR). We could demonstrate that our elephant isolate collection contained at least six different lineages that were representative for their source of origin. Despite generally broad antimicrobial susceptibility of S. agalactiae, many showed tetracycline resistance in vitro. S. agalactiae plays an important role in bacterial infections not only in cattle and humans, but also in elephants. Comparative studies were able to differentiate S. agalactiae isolates from elephants into different infectious clusters based on their epidemiological background.

Eisenberg, T., S. Poignant, Y. Jouan, A. Fawzy, W. Nicklas, C. Ewers, L. Mereghetti, and A. Guillon, "Acute Tetraplegia Caused by Rat Bite Fever in Snake Keeper and Transmission of Streptobacillus moniliformis.", Emerging infectious diseases, vol. 23, issue 4, pp. 719-721, 2017 04. Abstract

We report acute tetraplegia caused by rat bite fever in a 59-year old man (snake keeper) and transmission of Streptobacillus moniliformis. We found an identical characteristic bacterial pattern in rat and human samples, which validated genotyping-based evidence for infection with the same strain, and identified diagnostic difficulties concerning infection with this microorganism.

Noll, I., T. Eisenberg, K. Failing, M. Rohde, K. Schlez, W. Wolter, A. Fawzy, and M. Zschöck, "Untersuchung zur Verbreitung von Mycobacterium avium ssp. paratuberculosis (MAP) in hessischen Milchviehbeständen", Berliner und Münchener Tierärztliche Wochenschrift , vol. 130, pp. 13-20, 2017.
2016
Fawzy, A., A. Fayed, H. Youssef, A. El-Sayed, and M. Zschöck, "First report of MIRU-VNTR genotyping of Mycobacterium avium subsp. paratuberculosis isolates from Egypt.", Iranian journal of veterinary research, vol. 17, issue 2, pp. 130-133, 2016 Spring. Abstract

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne's disease, an economically important disease in ruminants worldwide. It was first isolated in Egypt in 2005. Since then, the pathogen has been detected in different Egyptian provinces. In order to trace the source of infection, genotyping using simple methods of high discriminatory power such as mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTR) were carried out in different countries. Until now there is no published information about MIRU-VNTR genotyping of MAP isolates in Egypt. To address that point, 100 faecal samples were collected and cultivated from 3 different suspected dairy farms. Fourteen isolates belonging to one farm were identified as MAP and subjected to genotyping using 8 different MIRU-VNTR loci PCRs. Two different genotypes were recognized based on size polymorphism observed in one locus (VNTR-7) that was confirmed by sequencing. Our work provides a preliminary basis of constructing a MIRU-VNTR genotyping database of MAP in Egypt.

Eisenberg, T., A. Fawzy, W. Nicklas, T. Semmler, and C. Ewers, "Phylogenetic and comparative genomics of the family Leptotrichiaceae and introduction of a novel fingerprinting MLVA for Streptobacillus moniliformis.", BMC genomics, vol. 17, issue 1, pp. 864, 2016 Nov 03. Abstract

BACKGROUND: The Leptotrichiaceae are a family of fairly unnoticed bacteria containing both microbiota on mucous membranes as well as significant pathogens such as Streptobacillus moniliformis, the causative organism of streptobacillary rat bite fever. Comprehensive genomic studies in members of this family have so far not been carried out. We aimed to analyze 47 genomes from 20 different member species to illuminate phylogenetic aspects, as well as genomic and discriminatory properties.

RESULTS: Our data provide a novel and reliable basis of support for previously established phylogeny from this group and give a deeper insight into characteristics of genome structure and gene functions. Full genome analyses revealed that most S. moniliformis strains under study form a heterogeneous population without any significant clustering. Analysis of infra-species variability for this highly pathogenic rat bite fever organism led to the detection of three specific variable number tandem analysis loci with high discriminatory power.

CONCLUSIONS: This highly useful and economical tool can be directly employed in clinical samples without laborious prior cultivation. Our and prospective case-specific data can now easily be compared by using a newly established MLVA database in order to gain a better insight into the epidemiology of this presumably under-reported zoonosis.

Fawzy, A., M. Zschöck, C. Ewers, and T. Eisenberg, "New polymorphisms within the variable number tandem repeat (VNTR) 7 locus of Mycobacterium avium subsp. paratuberculosis.", Molecular and cellular probes, vol. 30, issue 3, pp. 132-7, 2016 Jun. Abstract

Variable number tandem repeat (VNTR) is a frequently employed typing method of Mycobacterium avium paratuberculosis (MAP) isolates. Based on whole genome sequencing in a previous study, allelic diversity at some VNTR loci seems to over- or under-estimate the actual phylogenetic variance among isolates. Interestingly, two closely related isolates on one farm showed polymorphism at the VNTR 7 locus, raising concerns about the misleading role that it might play in genotyping. We aimed to investigate the underlying basis of VNTR 7-polymorphism by analyzing sequence data for published genomes and field isolates of MAP and other M. avium complex (MAC) members. In contrast to MAP strains from cattle, strains from sheep displayed an "imperfect" repeat within VNTR 7, which was identical to respective allele types in other MAC genomes. Subspecies- and strain-specific single nucleotide polymorphisms (SNPs) and two novel (16 and 56 bp) repeats were detected. Given the combination of the three existing repeats, there are at least five different patterns for VNTR 7. The present findings highlight a higher polymorphism and probable instability of VNTR 7 locus that needs to be considered and challenged in future studies. Until then, sequencing of this locus in future studies is important to correctly assign the underlying allele types.(1).

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