Mycobacterium tuberculosis exposure of livestock in a German dairy farm: implications for intra vitam diagnosis of bovine tuberculosis in an officially tuberculosis-free country.

Germany has been an officially bovine tuberculosis (bTB)-free (OTF) country since 1996. Gradually rising numbers of bTB herd incidents due to Mycobacterium bovis and M. caprae in North-Western and Southern Germany during the last few years prompted the competent authorities to conduct a nationwide bTB survey in 2013/2014. This led to the detection of a dairy herd in which as many as 55 cattle reacted positively to consecutive intra vitam testing. Test-positive animals lacked visible lesions indicative of bTB at necropsy. Extensive mycobacterial culturing as well as molecular testing of samples from 11 tissues for members of the M. tuberculosis complex (MTC) yielded negative results throughout. However, caseous lymphadenitis of Ln. mandibularis accessorius was observed during meat inspection of a fattening pig from the same farm at regular slaughter at that time. Respective tissue samples tested MTC positive by polymerase chain reaction, and M. tuberculosis T1 family were identified by spoligotyping. Four human reactors within the farmer's family were also found to be immunoreactive. As exposure of livestock to M. tuberculosis is not generally considered, its impact may result in regulatory and practical difficulties when using protocols designed to detect classical bTB, particularly in OTF countries.

Pattern of enterotoxin genes seg, seh, sei and sej positive Staphylococcus aureus isolated from bovine mastitis.

PCR detection of the genes encoding the newly described staphylococcal enterotoxins (SE) SEG, SEH, SEI and SEJ was carried out for 104 randomly selected Staphylococcus aureus field strains isolated from cases of bovine mastitis. Sixty-one (58.7%) isolates were positive for one or more of these novel enterotoxin genes. Thirty-six field strains were classified as carrier of seg, 22 of sei gene and 23 were positive for sej gene. None of the 104 investigated ruminant S. aureus strains carried the seh gene. Thirty-seven of these S. aureus strains showed a combination of genes encoding enterotoxin types SEA to SEE or toxic shock syndrome toxin 1 (TSST 1). Thirteen cultures harboured only one, 28 two, 12 three and 8 four enterotoxin genes. Among the 61 S. aureus field strains 14 (23.0%) were positive for the genes encoding SEJ and SED and 10 (16.4%) isolates for those encoding SEG and SEI. Isolates harbouring the sed/sej genes were further characterized by macrorestriction analysis and pulsed-field-gelelectrophoresis (Pfge). Macrorestriction analysis revealed six patterns. Nine of these14 S. aureus isolates (64.3%) exhibited two patterns with a high degree of relationship (>80%).

[Coxiella burnetii as zoonotic pathogen with special regard to food hygiene]. / Coxiella bumetii als Zoonoseerreger unter besonderer Berücksichtigung der Lebensmittelhygiene.

In Hesse, Germany, bulk milk of farms producing raw milk cheese is examined by PCR for Coxiella burnetii yearly. In 2003 the pathogen has been detected unusually frequent. By means of two examples the hygienic measures are shown, which were initiated by the veterinary administration. To detect Coxiella burnetii means always the preoccupation with unsolved questions. It is particularly uncertain, whether there is a risk of oral infection for the human being. From the point of view of food hygiene, surveys are needed urgently to work out a risk assessment. Based on this a uniform risk management and a reasonable risk communication can be fixed.

Shiga-toxin-producing Escherichia coli in faeces of healthy dairy cows, sheep and goats: prevalence and virulence properties.

The prevalence of Shiga-toxin-producing Escherichia coli (STEC) in healthy dairy ruminants was investigated between 1996 and 1998 by a multiplex polymerase chain reaction (mPCR) technique. A total of 13 552 E. coli colonies from 726 cows, 28 sheep and 93 goats out of 112 randomly selected dairy farms in Hessia, Germany were analysed. STEC strains were recovered from 131 (18.0%) cows, nine (32.1%) sheep and 70 (75.3%) goats. Further characterization of the STEC isolates showed that 89 (0.66% of the investigated colonies) of animal field strains carried stx1 gene, 64 (0.47%) stx2 gene and 57 (0.42%) stx1 and stx2 gene. Sixty (93.8%) out of 64 stx2 field strains were harboured by cows. In contrast, 74 (83.1%) out of 89 stx1 dairy animal field strains were from ovine or caprine origin. Only 17 (8. 1%) stx-positive isolates (13 from cattle, three from sheep and only one from goat) were positive for eaeA gene. Eight (9.0%) of the stx1, five (7.8%) of the stx2 and four (7.0%) of the stx1/stx2 gene-positive field strains carried the eaeA gene. The prevalence of EHEC-haemolysin (EHEC-hlyA) gene sequence was 88.8% (79 isolates) of the stx1 and 68.8% (44 isolates) of the stx2 isolates. Out of 57 stx1- and stx2-positive field-strains, 34 (59.6%) carried the EHEC-hlyA gene. E. coli O serovars O:157 and O:111 were not found. Only one isolate was positive with O26 antiserum.

[Thermophilic Campylobacter from fecal samples of animals and their behavior in the ganglioside (GM1)-ELISA, Vero cell test and salt aggregation test]. / Thermophile Campylobacter aus Faecesproben von Tieren und deren Verhalten im Gangliosid (GM1)-ELISA, Vero-Zelltest und Salzaggregationstest.

From 397 fecal specimens from apparently healthy and from diarrheic pigs, dogs, cats and cattle 59 strains (= 15%) of thermophilic Campylobacter (C.) spp. were isolated by culture. 39 strains were identified as C. coli and 18 as C. jejuni whereas 2 isolates could not be classified. None of the strains was found to be positive for cytotoxic enterotoxin in the GM1-ELISA. In the Vero-cell test 5 isolates showed a cytotoxic effect. The salt aggregation test (SAT) for indicating cell surface hydrophobicity was positive with 24 strains (5 C. jejuni, 19 C. coli). A correlation of isolation results with clinical manifestation could not be observed.

Multifocal serous chorioretinitis in beagle dogs.

In the years 1988 and 1989, routine ophthalmological examinations of dogs from the company-owned beagle colony revealed a clinically inapparent chorioretinitis in 7.4 and 10% of the animals, respectively, as it has previously been described by Weisse et al. (1981). The alterations were seen mainly in the non-tapetal fundus, and they appeared more frequently in both eyes than in just one eye. Infection tests as well as virologic, bacteriologic and histopathologic investigations were performed in order to clarify the origin. A direct evidence of virus particles from processed ocular material by electron microscopy was not possible. Tests for growth on MDCK cells were negative. In bacteriologic tests, a gram positive, filiform, branched microorganism was isolated. The histopathologic findings in the subacute stage were a focal atrophy of the first retinal neuron and a focal proliferation of glia cells.

[Comparative studies of the detection of rotavirus in fecal samples of calves with diarrhea with the latex test "Slidex Rota-Kit 2" and electron microscopy]. / Vergleichende Untersuchung zum Nachweis von Rotavirus in Kotproben durchfallkranker Kälber mit dem Latextest "Slidex Rota-Kit 2" und dem Elektronenmikroskop.

Of 68 fecal samples from calves with diarrhoea which were tested for rotavirus with the latex agglutination test "Slidex Rota-Kit2" and by electron microscopy 33 samples were positive and 33 were negative with both tests respectively. Divergent results (latex test positive/EM negative and vice versa) were observed in one specimen only, respectively. Cross reactions with other viruses diagnosed by electron microscopy were not observed with the latex agglutination test. The "Slidex Rota-Kit2" is another suitable test for the diagnostic laboratory as well as for the veterinary practitioner for the detection of rotavirus in fecal samples of calves.

[Microbiological study results (bacteriology and electron microscopy) of diarrhea in dog whelps]. / Mikrobiologische Untersuchungsergebnisse (Bakteriologie und Elektronenmikroskopie) bei der Diarrhö des Hundewelpen.

Bacteriological examinations of 159 faeces and intestinal contents of dogs with diarrhoea revealed E. coli in 157 specimens. 73 of these samples contained non haemolytic strains, 18 haemolytic isolates, and 66 haemolytic, as well as non haemolytic strains. Klebsiella sp. and Staphylococcus aureus were found in 9 cases, and Salmonella sp. (group B) was isolated once only. By electron microscopy parvovirus could be detected in 19 samples. Ten were positive for coronavirus and one for rotavirus. Morphologically not finally identified coronavirus like and picornavirus like particles were found in 3 cases, respectively. A significant relationship between the occurrence of virus infections and the isolation of certain bacteria species was not found. In 45 E. coli strains virulence factors, such as the heat labile enterotoxin (LT) and the verotoxin (VT), could not be detected, but 31 of these isolates showed different haemagglutination patterns which were still present by 14 of them in the presence of mannose (mannose resistant haemagglutination, MR-HA). These MR-HA inducing E. coli isolates were present more often in parvovirus positive samples (in 6 of 10) than they could be detected in parvovirus negative ones (in 8 of 35), implicating a potential pathogenic role of these E. coli strains for the parvovirus enteritis of dogs.

[Microbiological development studies in a clinically healthy, closed primate colony (Macaca mulatta, Macaca arctoides) after the establishment of a Salmonella infection]. / Mikrobiologische Verlaufsuntersuchungen in einer klinisch gesunden, geschlossenen Primatenkolonie (Macaca mulatta, Macaca arctoides) nach Feststellung einer Salmonellose.

A case of a Salmonella infantis septicemia in a primate (Macaca mulatta) probably induced by shipment stress gave rise to microbiological examination of fecal samples from all animals of a since 1976 closed colony of Macaca mulatta and Macaca arctoides. During 3 investigations carried out in 3 week intervals salmonella-positive animals were separated immediately from the stock. At the first examination 3 of 52, and at the second 2 of 49 samples were found to be salmonella-positive. At the third series all of the remaining 47 animals proved to be salmonella-negative. All isolates were identified as S. infantis. Feeding of contaminated, dried prawns was suspected as the most probable source of infection. A simultaneous examination of 96 of the stool specimens revealed Y. enterocolitica in 4 samples. Additionally, electron-microscopy of pooled fecal samples (3 to 4 animals each) led to demonstration of corona-, adenovirus and small round virus-like particles in one pool each. Moreover, coronavirus-like particles could be detected in 7 of 13 (1st examination series), 5 of 15 (2nd series) and 1 of 13 pools (3rd series).