Gastrointestinal parasite assemblages from the wild rodent capybara (Hydrochoerus hydrochaeris) inhabiting a natural protected area from Argentina.

Knowledge about parasitic diseases of wildlife will help us to understand the dynamics of parasites and their effects on host populations. The capybara (Hydrochoerus hydrochaeris) is the largest living rodent in the world, and its distribution is associated with the presence of tropical and subtropical wetlands in South America. The Los Padres Lake Integral Reserve (LPLIR) is an important conservation zone in the pampean region of Argentina. One of the emblematic species found within the reserve is the capybara. The objective of this study was to determine the gastrointestinal parasites present in wild capybaras of the LPLIR and to compare different coprological methodologies. Free-ranging capybara fresh feces from 57 individuals were randomly collected from the area of LPLIR in the summer of 2022. Three different techniques were applied: spontaneous sedimentation technique (SS), INTA modified McMaster technique (MM), and Mini-FLOTAC (MF) technique. Fifty-six samples from all samples analysed (56/57, 98%) were found to be positive for gastrointestinal parasites. Two species of Strongylida, Protozoophaga obesa, Echinocoleus hydrochaeris, one unidentified nematode, one unidentified spirurid, and at least two morphotypes of Eimeria spp. oocysts were recorded. There were found significant differences in the proportion of positive samples and in richness by technique, but no significant differences were found in parasite counting. In conclusion, the choice of methodology depends on the specific objectives of the study. This is the first parasitological study of capybaras from the LPLIR and represents an exploration of parasite communities present in these wild rodents at their southernmost distribution.

Associations between host characteristics and antimicrobial resistance of Salmonella typhimurium.

A collection of Salmonella Typhimurium isolates obtained from sporadic salmonellosis cases in humans from Lower Saxony, Germany between June 2008 and May 2010 was used to perform an exploratory risk-factor analysis on antimicrobial resistance (AMR) using comprehensive host information on sociodemographic attributes, medical history, food habits and animal contact. Multivariate resistance profiles of minimum inhibitory concentrations for 13 antimicrobial agents were analysed using a non-parametric approach with multifactorial models adjusted for phage types. Statistically significant associations were observed for consumption of antimicrobial agents, region type and three factors on egg-purchasing behaviour, indicating that besides antimicrobial use the proximity to other community members, health consciousness and other lifestyle-related attributes may play a role in the dissemination of resistances. Furthermore, a statistically significant increase in AMR from the first study year to the second year was observed.

Use of multilocus variable-number tandem repeat analysis (MLVA) in eight European countries, 2012.

Genotyping of important medical or veterinary prokaryotes has become a very important tool during the last decades. Rapid development of fragment-separation and sequencing technologies has made many new genotyping strategies possible. Among these new methods is multilocus variable-number tandem repeat analysis (MLVA). Here we present an update on the use of MLVA in eight European countries (Denmark, France, Germany, Ireland, Italy, the Netherlands, Norway and Sweden). Researchers in Europe have been active in developing and implementing a large array of different assays. MLVA has been used as a typing tool in several contexts, from aiding in resolving outbreaks of foodborne bacteria to typing organisms that may pose a bioterrorist threat, as well as in scientific studies.

[Role of pathogen surveillance and subtyping in outbreak recognition of food-borne bacterial infections. Microbiological perspective - Aims, methods and prospects of pathogen subtyping]. / Rolle der Erregersurveillance und -subtypisierung zur Ausbrucherkennung bei lebensmittelbedingten bakteriellen Infektionen. Sicht der Mikrobiologie - Ziele, Methoden und Perspektiven der Erregersubtypisierung.

The recognition of infection clusters via determination of clonal relationships between pathogen isolates represents the major aim of pathogen subtyping during outbreaks. In addition, a continuing and comprehensive subtyping of pathogen isolates is a prerequisite for early recognition of changes within pathogen populations, especially of new pathogen types and variants. Here, in an exemplary manner, we outline the current practice in Germany for three important agents of food-borne infections, Salmonella enterica, Listeria monocytogenes and enterohemorrhagic Escherichia coli (EHEC). Pathogen subtyping is mostly performed in specialized laboratories. Collection of representative pathogen isolates is therefore critical for comprehensive pathogen surveillance. Salmonella and L. monocytogenes are usually isolated by sample culturing in primary diagnostic laboratories and a considerable number are sent to the respective reference laboratories for further subtyping. However, the current situation in terms of EHEC is problematic. As the detection of shiga toxin (or gene) is sufficient for diagnosis and case reporting, primary diagnostic laboratories actually rarely isolate EHEC; therefore, a concept for appropriate retrieval of isolates is needed to ensure effective EHEC surveillance in Germany.

A quantitative approach to analyse linkages between antimicrobial resistance properties in Salmonella Typhimurium isolates.

This study used statistical methods to investigate linkages in phenotypic resistance profiles in a population sample of 321 Salmonella Typhimurium isolates from sporadic salmonellosis cases in Lower Saxony, Germany, collected during 2008-2010. A resistance index was applied to calculate the conditional probability of resistance to one antimicrobial agent given the resistance to one or more other antimicrobial agent(s). A susceptibility index was defined analogously. A contingency plot, which visualizes the association between resistances to two antimicrobial agents, facilitated the interpretation. Linkages between minimum inhibitory concentrations (MIC) were analysed using Spearman's rank correlation coefficient and jittered scatter plots. Applying these methods provided a compact description of multi-resistance and linkages between resistance properties in large datasets. Moreover, this approach will improve monitoring of antimicrobial resistance dynamics of bacteria in human or animal populations by identifying linked resistance to antimicrobial agents (cross- or co-resistance) with a non-molecular method.

Comparative biology of IncQ and IncQ-like plasmids.

Plasmids belonging to Escherichia coli incompatibility group Q are relatively small (approximately 5 to 15 kb) and able to replicate in a remarkably broad range of bacterial hosts. These include gram-positive bacteria such as Brevibacterium and Mycobacterium and gram-negative bacteria such as Agrobacterium, Desulfovibrio, and cyanobacteria. These plasmids are mobilized by several self-transmissible plasmids into an even more diverse range of organisms including yeasts, plants, and animal cells. IncQ plasmids are thus highly promiscuous. Recently, several IncQ-like plasmids have been isolated from bacteria found in environments as diverse as piggery manure and highly acidic commercial mineral biooxidation plants. These IncQ-like plasmids belong to different incompatibility groups but have similar broad-host-range replicons and mobilization properties to the IncQ plasmids. This review covers the ecology, classification, and evolution of IncQ and IncQ-like plasmids.

Nucleotide sequence of the bacterial streptothricin resistance gene sat3.

The nucleotide sequence of the sat3 gene which encodes resistance of enteric bacteria to the antibiotic streptothricin is reported. A protein with a molecular mass of about 23 kDa is expressed from this gene. The sat3 gene is not obviously related to any one of the streptothricin resistance determinants identified so far among Gram-negative or Gram-positive bacteria.

Epidemiological analysis of the dynamic and diversity of Salmonella spp. in five German pig production clusters using pheno- and genotyping methods: an exploratory study.

An exploratory study in five conventional pig production clusters was carried out to investigate the dynamic and diversity of Salmonella spp. within different production stages and sample site categories (pooled feces, direct and non-direct environment). Observing two production cycles per production cluster, a total of 1276 samples were collected along the pig production chain. Following a microbiological examination via culture, 2246 subcultures were generated out of 285 Salmonella positive samples and analysed by pheno- and genotyping methods. Based on a combination of serotyping, MLVA (multiple-locus variable-number tandem repeat (VNTR) analysis), PFGE (pulse-field gel electrophoresis) and MLST (multilocus sequence typing), an amount of 22.3% Salmonella positive samples were characterized in clonal lineages and its variants. Within each production cluster, one main clonal lineage could be identified and persisted over both production cycles with a large diversity of variants and a wide distribution in sample site categories and production stages. Results underline the importance of biosecurity with emphasis on the environment to prevent persistence and circulation of Salmonella within herds. Furthermore, the combined implementation of MLVA, PFGE and MLST with conventional culture techniques for isolate classification could be successfully applied as an effective and valuable tool for identifying similar pattern of Salmonella occurrence within pig production clusters.

Prevalence and deletion types of the pathogenicity island ETT2 among Escherichia coli strains from oedema disease and colibacillosis in pigs.

Piglet pathogenic Escherichia coli encoding Shigatoxin 2e and F18 adhesins are the etiological agents of oedema disease as well as of non-oedema disease colibacillosis. In order to reveal virulence differences among this pathogen, the presence of the pathogenicity island (PAI) E. coli type three secretion system 2 (ETT2) was examined. Using PCR and Southern blot techniques for the identification of the right, the middle, and the left region of this 29.9kb large genetic element, the entire ETT2 was found among E. coli O138:H(-), O139:H1, and O147:H6 strains originated from cases of oedema disease in Germany between 1995 and 2001 and belonging to various clonal types. In contrast, non-oedema disease E. coli isolates (e.g. O8:H19, 101:H(-), O141:H4) contain deleted subtypes of ETT2. These deletions cover the translocon part of the putative ETT2-encoded type III secretion apparatus. It is suggested that the entire ETT2 is associated with a particular virulence trait of piglet oedema disease E. coli (EDEC).

Streptothricin resistance as a novel selectable marker for transgenic plant cells.

Streptothricins are known as antimicrobial agents produced by Streptomyces spp. Bacterial resistance to streptothricin is mediated by specific enzymes exhibiting an acetyltransferase activity which renders the drug non-toxic for bacteria. The nucleotide sequence of several streptothricin resistance genes from bacteria have been described. Certain cells of eukaryotic parasites (such as Ustilago maydis or Leishmania spp.) are sensitive to streptothricin and the introduction of the bacterial resistance gene sat2 renders them resistant. We show that numerous species of plants are sensitive to low concentrations of streptothricin. Moreover, introduction of the bacterial resistance gene sat3 under the control of the 35S cauliflower mosaic virus promoter protects these cells from the toxic action of streptothricin. Therefore, sat3-mediated streptothricin resistance appears to be a promising selective marker for genetic manipulation of plant cells.

[Localization of insertion into E. coli chromosome of Tn7-like transposons controlling resistance to trimethoprim and streptothricin]. / Lokalizatsiia vnedreniia v khromosomu E. coli Tn7-podobnykh transpozonov, kontroliruiushchikh ustoichivost' k trimetoprimu i streptotritsinu.

To localize the insertion sites for Tn7-like transposons Tn1824, Tn1825 and Tn1826 the EcoRI-cleaved fragments of E. coli recA+ and recA- strains chromosome carrying the transposons were hybridized. It was shown that transposition of Tn7-like elements takes place in the strictly defined region of E. coli chromosome, like it had been previously described for transposon Tn7.

[Biotinylated DNA probe for detection of streptotricin acetylation genes]. / Biotinilirovannyi DNK-zond dlia obnaruzheniia genov atsetilirovaniia streptotritsina.

The biotin-labelled DNA probe for identification of functioning and silent genes for streptotricin acetylation has been constructed. The probe is homologous to sat1 gene of the movable genetic element Tn1825. The simplified modification of the hybridization technique using the biotin-labelled DNA probe is described.

[Characteristics of transposition of Tn7-like elements determining resistance to trimethoprim and streptomycin]. / Kharakteristika transpozitsii Tn7-podobnykh élementov, opredeliaiushchikh ustoichivost' k trimetoprimu i streptotritsinu.

The transposing elements Tn7, Tn1824, controlling the resistance to trimethoprim and Tn1925, Tn1826, carrying the streptothricin resistance genes were classified as a new transposon family on the basis of their physical structure. The comparative genetic analysis of the frequency, specificity and insertion orientation in different replicons, obtained in independent research systems in this study, demonstrated the identity of transposition characteristics of the transposons. The latter makes it possible to classify them as an independent transposon family. The peculiar feature of the Tn7-like elements family is their RecA-dependent transposition into the chromosome of Escherichia coli stimulated by bacteriophage Plkc transduction of the transposons.

[Evaluation of the possibility of using genetic and microbiological research methods for resolving current problems in the epidemiology of salmonellosis]. / Otsenka vozmozhnosti ispol'zovaniia geneticheskikh i mikrobiologicheskikh metodov issledovaniia dlia resheniia aktual'nykh voprosov épidemiologii sal'monellezov.

S. typhimurium strains isolated in 14 regions of the USSR have, parallel with considerable similarity in their biological characteristics, a number of essential differences. These differences become manifest in the determination of the plasmid spectra of the above organisms. The transfer of R-plasmid PLE518 F1 me fin+ with a molecular weight of 96 Md to strains, sensitive to the action of typing bacteriophages, renders these strains resistant to the lytic action of a number of phages, which leads to the conversion of their phage type. In some cases it may deteriorate the validity of the method of phage typing, used for epidemiological purposes.

Nucleotide sequence and genetic characterization of the novel IncQ-like plasmid pIE1107.

The analysis of the complete nucleotide sequence of the small resistance plasmid pIE1107 revealed a close similarity to the well-known IncQ plasmids. Highly conserved replication proteins and nearly identical origins of replication (oriV) suggest equivalent functions in the related replication systems. However, pIE1107 contains two copies of IncQ-oriV-like DNA which are slightly different regarding the iterons. Upon deletion of a silent copy of IncQ-oriV-like DNA the resulting plasmid is fully compatible with IncQ plasmids, indicating that there is no mutual communication between the replication control of the respective replicons. Experiments with cloned oriV DNA strongly suggest that the replication initiation protein of pIE1107 has specialized into the distinct target-iterons of its own oriV which differs only by a few nucleotides from the oriV of IncQ plasmids. Implications from the apparent highly specific protein-DNA recognition and from the incompatibility properties of pIE1107 for the evolution of a family of compatible, IncQ-like plasmids are discussed.

Genetic and molecular characterization of R plasmids incompatible with R387 (IncK).

More than 70 conjugative R plasmids have been isolated from wild-type strains originating mainly in South-East Europe and identified as incompatible with the reference plasmid R387 of the incompatibility group K. These plasmids, governing different resistance patterns, have been characterized as P1 cotransducible species of between 50 and 60 mega-daltons. In contrast to the genetic similarity between all these plasmids and the reference type R387, their DNA revealed different digestion patterns after EcoRI treatment, although a number of common fragments could be identified. The molecular and genetic properties of these plasmid species demonstrate a phylogenetic relatedness.

Temperature-dependent expression of conjugation pili by IncM plasmid-harbouring bacteria: identification of plasmid-encoded regulatory functions.

A 1,3 kb DNA fragment of the IncM plasmid R446, if cloned in a multicopy plasmid, inhibits in trans the expression of conjugation pili by IncM plasmid-harbouring host bacteria as indicated by their insensitivity to the IncM pilus-dependent bacteriophage phi M (Iml, insensitivity to phi M-mediated lysis). Determination of the nucleotide sequence of this DNA fragment, the introduction of deletions an the analysis of transposon insertions reveal two determinants, imlA and imlB, responsible for the Iml phenotype. A stretch of 80 bp of DNA containing imlA and about 450 bp of adjacent DNA comprising imlB, together, bring about inhibition of the typical expression of conjugation pili at 30 degrees C. The introduction of a transposable promoter probe and the construction of respective lacZ fusions indicate transcription of complementary strands in vivo overlapping in the region comprising imlA and imlB. Moreover, the expression of reporter genes discloses temperature-dependent transcription of the imlA-imlB-region in one direction. A particular subfragment of the 1.3 kb IncM plasmid-derived DNA does not inhibit conjugation pilus expression at 30 degrees C but stimulates in trans the formation of pili at 42 degrees C to give rise to untypical sensitivity to phi M at 42 degrees C in addition to 30 degrees C. Other subfragments reveal vital interferences with IncM plasmid-harbouring host cells. The putative nature of the cloned determinants interfering with the normal expression of IncM plasmid DNA is discussed.

Plasmid pattern analysis of natural bacterial isolates and its epidemiological implication.

Natural isolates of Escherichia coli, Salmonella typhimurium, Klebsiella pneumoniae, and Providencia stuartii were analysed to determine their plasmid content. This data allowed the identification of nosocomial strains of K. pneumoniae and P. stuartii and helped in the differentiation of epidemic strains of E. coli 0111 and S. typhimurium. Phenotypically similar isolates of S. typhimurium could be shown to be of independent origin using plasmid pattern analysis. The dissemination of a particular plasmid through different strains of S. typhimurium resulted in a simulation of a very widely distributed epidemic strain, because the plasmid interfered with the phage type of its host strain in addition to determining resistance properties. Plasmid pattern analysis disclosed two independently existing but interacting epidemic processes: a bacterial 'epidemic' strain may become disseminated over a large territory and may predominate there for a long time; a single plasmid, however, may also become distributed through many different bacterial strains and may spread over a large territory. Plasmid pattern analysis provides a valuable and universal epidemiological laboratory method.

Characterization of conjugative plasmids belonging to a new incompatibility group (IncZ).

More than 30 conjugative R plasmids between 60 and 70 Md in size were identified in wild type strains of Enterobacteriaceae isolated in German Democratic Republic, Bulgaria, Mongolia, Poland, Ethiopia, Iraque, and Soviet Union. They have been characterized by means of several genetic and molecular techniques as members of a new incompatibility group, termed IncZ. The properties of these plasmids including digestion pattern after EcoRI treatment demonstrate a phylogenetic relatedness.

The trimethoprim resistance transposon Tn7 contains a cryptic streptothricin resistance gene.

The transposon Tn7 codes for a trimethoprim resistance and for a streptomycin/spectinomycin resistance function of the bacterial host cells. Cloning of a restriction fragment of Tn7 into the vector plasmid pUC19 reveals the presence in Tn7 of an additional potential resistance determinant. A streptothricin resistance gene, which appears cryptic in the original Tn7 context becomes activated in the recombinant plasmid upon supplying the promoter function of the lacZ system of pUC19. These results together with previously published sequence data further disclose the modular character in the resistance gene regions of Tn7-like transposons.