Presence of Clostridium difficile in poultry and poultry meat in Egypt

C. difficile has been recognized as a potential zoonotic agent encouraging investigations of C. difficile prevalence and ribotypes in animals. Here we report the prevalence and diversity of Egyptian C. difficile in I) samples from healthy poultry (n = 50), II) samples from diseased poultry (n = 54), and III) poultry meat (n = 150). Thirteen isolates were obtained from seven healthy and five diseased animals, but no C. difficile was cultured from poultry meat. The isolated C. difficile strains belonged to 3 different PCR-ribotypes (039/2, 205 and 001/FLI01). The detection of strains related to RT 001 known for its ability to cause disease in humans makes poultry a potential reservoir for pathogenic C. difficile

ЯКІВ ДМИТРОВИЧ ГРАХОВ – ДІЯЧ КУЛЬТУРИ ТА ОСВІТИ В КАТЕРИНОСЛАВСЬКІЙ ГУБЕРНІЇ

Розглянуто життєвий шлях, освітянську, наукову та музеєзнавчу діяльність директора Катеринославської гімназії та училищ краю Якова Дмитровича Грахова в середині ХІХ ст.The article covers life, scientific and museum activities of Yakiv Dmitrovych Grakhov, a director of Katerinoslav schools in the middle of the 19th centuary

Комплексный подход к выбору технологии разработки залежей тяжелых углеводородов

Целью работы является: выбор и технологическое обоснование разработки залежей тяжелых углеводородов. В выпускной квалификационной работе были рассмотрены сведения о тяжелых углеводородах: географическое распределение, геологические особенности строения пластов с высоковязкими нефтями и физико-химические характеристики тяжелых углеводородов. Были изучены технологии разработки залежей высоковязких нефтей и природных битумов, выделены критерии применения методов увеличения нефтеотдачи, рассмотрено используемое оборудования и выбор сетки скважин в зависимости от геологических условий. Проанализированы техника и технология методов добычи высоковязкой нефти, критерии их применимости, а также выделены достоинства и недостатки каждого метода. Область применения: месторождения высоковязких нефтей.The purpose of the work is: selection and technological justification for the development of heavy hydrocarbon deposits. In the final qualifying work, information about heavy hydrocarbons was considered: geographical distribution, geological features of the structure of reservoirs with high-viscosity oils, and physical and chemical characteristics of heavy hydrocarbons. Technologies for the development of deposits of high-viscosity oils and natural bitumen were studied, criteria for the use of methods for increasing oil recovery were identified, the equipment used and the choice of the well grid depending on geological conditions were considered. The technique and technology of high-viscosity oil production methods, the criteria for their applicability, and the advantages and disadvantage

Clostridioides difficile in South American Camelids in Germany: First Insights into Molecular and Genetic Characteristics and Antimicrobial Resistance

Little is known about zoonotic pathogens and their antimicrobial resistance in South American camelids (SAC) in Germany including Clostridioides (C.) difficile. The aim of this study was to investigate prevalence, molecular characteristics and antimicrobial resistance of C. difficile in SAC. Composite SAC faecal samples were collected in 43 husbandries in Central Germany and cultured for C. difficile. Toxinotyping and ribotyping was done by PCR. Whole genome sequencing was performed with Illumina® Miseq™. The genomes were screened for antimicrobial resistance determinants. Genetic relatedness of the isolates was investigated using core genome multi locus sequence typing (cgMLST) and single nucleotide polymorphism analysis. Antimicrobial susceptibility testing was done using the Etest® method. Eight C. difficile isolates were recovered from seven farms. The isolates belonged to different PCR ribotypes. All isolates were toxinogenic. cgMLST revealed a cluster containing isolates recovered from different farms. Seven isolates showed similar resistance gene patterns. Different phenotypic resistance patterns were found. Agreement between phenotypic and genotypic resistance was identified only in some cases. Consequently, SAC may act as a reservoir for C. difficile. Thus, SAC may pose a risk regarding zoonotic transmission of toxinogenic, potentially human-pathogenic and resistant C. difficile isolates

Use of a Western blot technique for the serodiagnosis of glanders

<p>Abstract</p> <p>Background</p> <p>The in vivo diagnosis of glanders relies on the highly sensitive complement fixation test (CFT). Frequently observed false positive results are troublesome for veterinary authorities and cause financial losses to animal owners. Consequently, there is an urgent need to develop a test with high specificity. Hence, a Western blot assay making use of a partly purified lipopolysaccaride (LPS) containing antigen of three <it>Burkholderia mallei </it>strains was developed. The test was validated investigating a comprehensive set of positive and negative sera obtained from horses and mules from endemic and non endemic areas.</p> <p>Results</p> <p>The developed Western blot assay showed a markedly higher diagnostic specificity when compared to the prescribed CFT and therefore can be used as a confirmatory test. However, the CFT remains the test of choice for routine testing of glanders due to its high sensitivity, its feasibility using standard laboratory equipment and its worldwide distribution in diagnostic laboratories.</p> <p>Conclusions</p> <p>The CFT should be amended by the newly validated Western blot to increase the positive likelihood ratio of glanders serodiagnosis in non endemic areas or areas with low glanders prevalence. Its use for international trade of horses and mules should be implemented by the OIE.</p

Prevalence of Coxiella burnetii in clinically healthy German sheep flocks

<p>Abstract</p> <p>Background</p> <p>Current epidemiological data on the situation of <it>Coxiella (C.) burnetii </it>infections in sheep are missing, making risk assessment and the implementation of counteractive measures difficult. Using the German state of Thuringia as a model example, the estimated sero-, and antigen prevalence of <it>C. burnetii </it>(10% and 25%, respectively) was assessed at flock level in 39/252 randomly selected clinically healthy sheep flocks with more than 100 ewes and unknown abortion rate.</p> <p>Results</p> <p>The CHECKIT™ Q-fever Test Kit identified 11 (28%) antibody positive herds, whereas real-time PCR revealed the presence of <it>C. burnetii </it>DNA in 2 (5%) of the flocks. Multiple-locus variable number of tandem repeats analysis of 9 isolates obtained from one flock revealed identical profiles. All isolates contained the plasmid QpH1.</p> <p>Conclusions</p> <p>The results demonstrate that <it>C. burnetii </it>is present in clinically inconspicuous sheep flocks and sporadic flare-ups do occur as the notifications to the German animal disease reporting system show. Although <it>C. burnetii </it>infections are not a primary veterinary concern due to the lack of significant clinical impact on animal health (with the exception of goats), the eminent zoonotic risk for humans should not be underestimated. Therefore, strategies combining the interests of public and veterinary public health should include monitoring of flocks, the identification and culling of shedders as well as the administration of protective vaccines.</p

Die Rolle des Fettsäuresynthase Acyl-Carrier-Proteins (FAS-ACP) in der Chinoxalinantibiotika-Biosynthese

Chinoxalinantibiotika sind cyclische Octadepsipeptide, die aus zwei antiparallel angeordneten und über Esterbindungen miteinander verknüpften Chinoxalinoyl-tetrapeptidketten bestehen. Die Peptidketten sind an ihren Aminotermini mit Chinoxalin-2-carbonsäure (2QA) acyliert. Diese planaren, heteroaromatischen Seitengruppen sind für den Mechanismus der bifunktionellen Interkalation der Chinoxalinantibiotika in DNA verantwortlich. Als Inhibitoren der Transkription und der Replikation besitzen die Chinoxalin-antibiotika antimikrobielle und cytostatische Aktivität. Chinoxalinantibiotika werden von verschiedenen Streptomyces Arten, zu denen auch Streptomyces triostinicus und Streptomyces echinatus gehören, produziert. Der Aufbau der Peptidketten erfolgt in einem nichtribosomalen Mechanismus durch die Triostin Synthetasen I, II, III und IV (TrsI, II, III und IV). Die Triostin Synthetase I (TrsI) ist eine allein stehende Adenylierungs-Domäne (A-Domäne) und aktiviert die 2QA als Adenylat ohne 2QA-Bindungseigenschaft, während die Triostin Synthetasen II, III und IV typische nichtribosomale Peptidsynthetasen (NRPS) mit insgesamt vier kompletten Modulen sind, die jeweils eine Aminosäure des 2QA-Tetrapeptids durch Adenylierung aktivieren und als Thioester kovalent binden können. Die Peptidsynthetasen TrsII, III und IV sind somit höchstwahrscheinlich für den Aufbau der Tetrapeptidketten verantwortlich. Auf der Suche nach der 2QA-bindenden Proteinkomponente des Chinoxalinantibiotika-NRPS-Komplexes wurde aus Streptomyces echinatus ein ca. 9 kDa großes 2QA-bindendes Protein isoliert, welches durch die TrsI mit 2QA in ATP-abhängiger Weise acyliert wird. Die Reinigung und aminoterminale Sequenzierung des als AcpPSE bezeichneten 2QA-bindenden Proteins sowie die anschließende Klonierung seines Gens (acpPSE) ergab, dass es sich bei diesem Protein um das Acyl-Carrier-Protein (ACP) aus der Fettsäuresynthase (FAS) von S. echinatus handelt. Die heterologe Expression von acpPSE in E. coli lieferte das rekombinante AcpPSE in seiner holo-Form. Die eingehende Charakterisierung des ACPs ergab, dass es von ähnlichen allein stehenden A-Domänen (ACMSI und MKMSI aus der Actinomycin- bzw. Etamycin-Biosynthese) verwandter Chromopeptid-Biosynthesesysteme aus Streptomyceten nicht acyliert wird und ausschließlich ein Substrat für die TrsI darstellt. Aus diesen Daten konnte geschlossen werden, dass es sich bei der TrsI und des AcpPSEs höchstwahrscheinlich um das Initiationsmodul der Chinoxalinantibiotika-Biosynthese handelt, welches Chinoxalin-2-carbonsäure für die Synthese des 2QA-Tetrapeptids bereitstellt. Die Bestimmung der kinetischen Parameter KM und kcat der Acylierungsreaktion für das putative Initiationsmodul TrsI/AcpPSE und der Vergleich mit Initiationsmodulen anderer Chromopeptid-Biosynthese-systeme (Actinomycin-, Enterobactin- und Vibriobactin-Biosynthese) zeigten, dass TrsI/AcpPSE eine typische Acylierungs-Kinetik für diesen Modul-Typ aufweist. Die Umsetzung des Proteinpaares TrsI/AcpPSE mit der Triostin Synthetase II, welche die Module 2 (Serin) und 3 (Alanin) für den Aufbau der Tetrapeptidketten enthält, zeigte, dass AcpPSE spezifisch 2QA auf das an TrsII kovalent gebundene Serin unter Bildung von 2QA-Serin überträgt. Zudem war die TrsII offensichtlich in der Lage, das 2QA-L-Serine in 2QA-D-Serine durch Epimerisierung umzuwandeln. Diese Daten zeigen, dass AcpPSE eine hochspezifische Bindungsdomäne für 2QA in der Initiationsreaktion der 2QA-Peptidsynthese darstellt und hiermit zum ersten Mal die Mitwirkung eines Acyl-Carrier-Proteins der bakteriellen Fettsäuresynthase an einer nichtribosomalen Peptidsynthese gezeigt wird

Multidrug-resistant Clostridium Difficile Ribotypes 078 and 014/5-FLI01 in Piglets From Costa Rica

Though an overlap of Clostridium difficile PCR ribotypes (RT) in humans and animals has been noted -particularly in piglets-information regarding C. difficile isolates from swine is scarce in Latin America. A characterization of 10 C. difficile isolates obtained from this origin in Costa Rica revealed the presence of the RT078 (n = 4) and RT014/5-FLI01 (n = 6) ribotypes. Unlike two previous reports from the region, all isolates were multidrug resistant (MDR). According to a minimum spanning tree (MST) analysis, our RT078 isolates formed a clonal complex with some German RT078 isolates and the already noted overlap of RT078 strains in humans and animals. This unanticipated high level of genetic relatedness confirms the transcontinental spread and geographically unlimited clustering of RT078.Aunque se ha observado una superposición de ribotipos de PCR de Clostridium difficile (RT) en seres humanos y animales -en particular en lechones-, la información relativa a los ribotipos de C. difficile aislados de los cerdos es escasa en América Latina. La caracterización de los difficileisolatos de 10C. obtenidos de este origen en Costa Rica reveló la presencia de los ribotipos RT078 (n¼4) y RT014/5-FLI01 (n¼6). A diferencia de dos informes anteriores de la región, los alisolados eran multiresistentes a las drogas (MDR). Según un análisis de árbol de expansión mínima (MST), nuestros aislados RT078 formaron un complejo clonal con algunos aislados alemanes RT078 y la ya mencionada superposición de cepas RT078 en humanos y animales. Este alto nivel imprevisto de relación genética confirma la propagación transcontinental y la agrupación geográficamente ilimitada de RT078.Escuela Medicina Veterinari

<i>Clostridioides difficile</i> in South American Camelids in Germany: First Insights into Molecular and Genetic Characteristics and Antimicrobial Resistance

Little is known about zoonotic pathogens and their antimicrobial resistance in South American camelids (SAC) in Germany including Clostridioides (C.) difficile. The aim of this study was to investigate prevalence, molecular characteristics and antimicrobial resistance of C. difficile in SAC. Composite SAC faecal samples were collected in 43 husbandries in Central Germany and cultured for C. difficile. Toxinotyping and ribotyping was done by PCR. Whole genome sequencing was performed with Illumina® Miseq™. The genomes were screened for antimicrobial resistance determinants. Genetic relatedness of the isolates was investigated using core genome multi locus sequence typing (cgMLST) and single nucleotide polymorphism analysis. Antimicrobial susceptibility testing was done using the Etest® method. Eight C. difficile isolates were recovered from seven farms. The isolates belonged to different PCR ribotypes. All isolates were toxinogenic. cgMLST revealed a cluster containing isolates recovered from different farms. Seven isolates showed similar resistance gene patterns. Different phenotypic resistance patterns were found. Agreement between phenotypic and genotypic resistance was identified only in some cases. Consequently, SAC may act as a reservoir for C. difficile. Thus, SAC may pose a risk regarding zoonotic transmission of toxinogenic, potentially human-pathogenic and resistant C. difficile isolates