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Infections caused by species of the genus Mannheimia cause diverse disease complexes in many wild and domestic animals worldwide. Fast and accurate detection of single species within the genus remains an unsolved problem till today. To resolve this diagnostic challenge, we developed a real-time PCR assay for the rapid and specific identification of five species of the genus Mannheimia (M. haemolytica, M. varigena, M. ruminalis, M. granulomatis and M. glucosida) from bacterial cultures and tissue samples. The assay was validated with reference strains, field isolates and bacteria spiked tissue samples. The sodA gene was used as target region for species-specific primer pairs. The real-time PCR assay demonstrated species specificity for all five examined Mannheimia spp. and a rapid test completion time of less than 5 h. This is a considerable advantage compared to the traditional phenotyping methods currently used to distinguish between the species of the genus. The assay was able to detect approximately 10(3) bacterial cells per gram lung tissue sample, as determined with spiked tissue samples. We assume that the assay could become useful for fast laboratory diagnostic assessment particularly of respiratory infections caused by Mannheimia in animals.

A total of 500 streptococci from two indications of swine (beta-haemolytic streptococci from infections of the urinary/genital tract including strains from the mastitis metritis agalactia syndrome as well as S. suis from infections of the central nervous system and the musculoskeletal system), two indications of horses (S. equi from respiratory tract infections and beta-haemolytic streptococci from infections of the genital tract), as well as three indications of dogs and cats (beta-haemolytic streptococci from infections of the respiratory tract, the urinary/genital tract, and skin/ear/mouth) were investigated for their susceptibility to antimicrobial agents. Regardless of the animal origin and indication, the most frequently detected resistance properties were resistances against sulfamethoxazole (20-78%), tetracycline (17-93%) as well as gentamicin (14-79%). Resistance to penicillins or cephalosporins was very rarely detected - if at all.

A total of 120 isolates of Klebsiella spp. and Proteus spp. collected from horses and small animals (dogs and cats) were screened for their susceptibility to 24 different antimicrobial agents. Klebsiella spp. were included from infections of the genital tract (GT) of horses (36 isolates) and the urinary/genital tract (UGT) from dogs and cats (17 isolates), while Proteus spp. were from small animal (dogs and cats) infections of the UGT (37 strains) and the skin (incl. ear/mouth) (30 isolates). In Klebsiella spp. resistance appeared most frequently to ampicillin (53-67%), sulfamethoxazole (19-29%) and potentiated sulfonamides (trimethoprim/sulfamethoxazole 1/19 combination) (19-24%). A further 29% of enrofloxacin resistant Klebsiella isolates were observed for the UGT of small animals. From the GT of horses for this antimicrobial agent there was no isolate detected with a comparably high minimum inhibitory concentration (MIC) value. In Proteus spp. highest percentages of resistance occurred against tetracycline (90-92%). Due to drug efflux proteins, high MIC values against this antimicrobial agent have been frequently reported in literature. In Proteus spp. relevant resistance percentages also occurred for potentiated sulfonamides (27-37%), sulfamethoxazole (24-37%) and chloramphenicol (24-37%).

To determine the current status of antimicrobial susceptibility of bacterial pathogens from animals in Germany, the Bff-GermVet monitoring program was initiated as a complementary program to the German national monitoring program GERM-Vet conducted by the Federal Office of Consumer Protection and Food Safety (BVL). In the Bff-GermVet program, a total of 1,626 bacterial strains, obtained during a 27-month period (01/2004-03/2006) from 31 indications, was screened for susceptibility against 22 antimicrobial agents and two combinations of antimicrobial agents. Selected bacteria were additionally tested for their susceptibility against additional three combinations of antimicrobial agents and the corresponding single substances. This paper describes the overall aims and the structure of the program with particular reference to the sampling strategy, the methodology for susceptibility testing and the interpretive criteria used for evaluation of the results.

A total of 417 isolates of Escherichia coli collected from five animal species/organ system combinations from swine [urinary/genital tract (UGT) incl. mastitis metritis agalactia syndrome], horses [genital tract (GT)] and dogs/cats [respiratory tract (RT), UGT and gastrointestinal tract (GIT)] were analysed quantitatively for their susceptibility against different antimicrobial agents by determination of minimum inhibitory concentrations. Regardless of which animal species the strains originated from, resistance appeared most frequently against sulfamethoxazole (18-59%), tetracycline (14-54 %), and ampicillin (14-39%). High percentages of intermediate isolates were observed for cephalothin (39-46 %). In general, low prevalences of resistance were detected for amoxicillin/clavulanic acid (1-4%), gentamicin (1-9%), and cefazolin (0-11%). Generally speaking, the antimicrobial resistance situation among E. coli isolates from horses and small animals is relatively good.

Objectives To discern the relevance of ST648 extended-spectrum β-lactamase (ESBL)-producing Escherichia coli as a putative new group of multiresistant and extraintestinal pathogenic strains in animals, its frequency, ESBL types, antimicrobial resistance patterns and virulence gene (VG) profiles should be determined and compared with ST131 strains from the same collection of strains. Methods ESBL-producing E. coli isolates (n = 1152), consecutively sampled from predominantly dogs, cats and horses between 2008 and 2011, were assigned to a phylogenetic group by PCR. Partial multilocus sequence typing was performed for group D and B2 strains and strains presumed to be D-ST648 and B2-ST131 were fully typed. ESBL genes and extraintestinal pathogenic E. coli (ExPEC)-like VGs were characterized by PCR and sequence analysis and antimicrobial resistance was determined by broth dilution. Clonal analysis was done by PFGE. Results Forty (3.5%) ESBL-producing E. coli were determined as D-ST648, whereas B2-ST131 isolates occurred less frequently (2.8%). Although the predominant ESBL type in both groups was CTX-M-15 (72.5% versus 46.9%), ST648 strains from companion animals and horses displayed a lower variety of ESBL types (CTX-M-1, -3, -14, -15 and -61 versus CTX-M-1,-2,-14,-15,-27 and -55 and SHV-12). In contrast to ST131 strains, a higher proportion of ST648 strains showed resistance to most non-β-lactam antibiotics. Overall, VGs were less abundant in ST648 strains, although some strains had VG profiles comparable to those of ST131 strains. ExPEC-associated serotype O1:H6 was predominant (46.8%) among the ST648 strains. Some PFGE clusters comprised ST648 isolates from pets, horses and wild birds and humans included from previous studies. Conclusions Our findings demonstrate that certain subgroups of E. coli D-ST648-CTX-M may represent a novel genotype that combines multiresistance, extraintestinal virulence and zoonotic potential. [ABSTRACT FROM PUBLISHER]

Antimicrobial resistance of Staphylococcus aureus is a major problem in human and veterinary medicine. The aim of this study was to characterise S. aureus isolates from wild and zoo animals mainly associated with bacterial infections. In total, 23 S. aureus isolates, including nine from wild animals and 14 from zoo animals, were obtained during routine diagnostics. All isolates were subjected to multilocus sequence typing (MLST), spa typing, macrorestriction analysis with subsequent SmaI pulsed-field gelelectrophoresis (PFGE), antimicrobial susceptibility testing and S. aureus -specific DNA-microarray analysis. Resistant isolates were also tested for their respective resistance genes by PCR. Isolates from zoo animals and wildlife showed a high diversity of MLST types, spa types and PFGE patterns. Nineteen different spa types were identified, including three novel types and 16 main macrorestriction patterns. Only few isolates were resistant to members of four classes of antimicrobial agents and harboured the respective resistance genes (β-lactams [ blaZ, mecA, mecC ], tetracyclines [ tet (K), tet (L)] and chloramphenicol [ cat pC221 ]) or mutations (fluoroquinolones). The DNA microarray analysis identified one isolate from a zoo animal harbouring the toxic shock syndrome toxin gene tst1 . Moreover, several enterotoxin genes were detected in five S. aureus isolates. All isolates were negative for Panton-Valentine leukocidin (PVL) genes, but the animal-associated leukocidin genes lukM / lukF-P83 were found in three isolates from two animals. [ABSTRACT FROM AUTHOR]