[Infectious agent detection in reproductive disorders in swine herds. Retrospective evaluation of diagnostic laboratory examinations]. / Erregernachweis bei Reproduktionsstörungen in Schweinebeständen. Retrospektive Auswertung labordiagnostischer Untersuchungen.

OBJECTIVE: The detection rate of various viral and bacterial agents causing reproductive failure in sows was analysed. MATERIAL AND METHODS: Samples from abortion/uterus (n=714), sera from weak born piglets (n=317), cervical swabs (n=881) and urogenital organs from slaughtered sows (n=416) that had been submitted between January 2006 and June 2009 were included in this analysis. Various PCR assays were run to detect PRRSV, PCV2, PPV, Chlamydia spp. and Leptospira spp. Other bacterial agents were examined using standard cultural methods. RESULTS: In material from abortion, detection rates of 14.7% for PCV2 and 6.8% for PRRSV EU genotype were revealed using PCR screening. The other agents were detected in single cases only (PPV 2.2%, PRRSV US genotype 1.8%, Chlamydia spp. 1.0%, Leptospira spp. 0.8%). Single PCR yielded a significantly higher detection rate for PCV2 than PCR screening. Comparing results from abortion/uterus and serum samples from weak born piglets, a significantly higher detection rate of PCV2 and PRRSV was found in sera. Bacteriological examination revealed bacterial agents in more than 75% of all cervical swabs. However, half of them had to be considered as contaminated due to the diversity of the isolated bacteria. Bacteriological testing of urogenital organs from slaughtered sows yielded positive results in 60% of all samples, with a remarkably lower proportion of contaminated samples of 7.4%. CONCLUSION: It is assumed that 60-70% of all cases of reproductive failure are similarly not caused by primary infections. If PRRSV infection is suspected, examination of serum samples from weak born piglets is much better than the testing of foetuses or other material from abortion. Due to poor detection rates, the examination of foetuses/abortion material by screening PCR cannot be recommended. In the case of bacterial infections of the urogenital system, the cultural examination of organs from slaughtered sows is more often successful than the testing of cervical swabs.

Occurrence of Mycoplasma hyopneumoniae infections in suckling and nursery pigs in a region of high pig density.

The occurrence of Mycoplasma hyopneumoniae infections in young pigs was surveyed in a retrospective study of 1122 datasets obtained from routine diagnostics where either suckling or nursery pigs were examined for M hyopneumoniae in lung tissue. Findings were correlated with the presence of lung lesions, detection of other respiratory pathogens, vaccination history and parameters describing the herd of origin. The prevalence of M hyopneumoniae in lung tissue from 201 suckling pigs was 2.0 per cent and, therefore, significantly lower than in lung tissue from 921 nursery pigs, which was 9.3 per cent. Previous use of antimicrobials and the vital status of the pigs when delivered for postmortem examination did not influence the detection of M hyopneumoniae infection. The presence of the porcine reproductive and respiratory syndrome virus (PRRSV)-EU genotype, Pasteurella multocida, Haemophilus parasuis, Mycoplasma hyorhinis or Streptococcus suis was correlated with a higher probability of also finding M hyopneumoniae. The history of vaccination, the time of the first or second application, and the type of vaccine (one- versus two-shot) did not influence the detection of M hyopneumoniae. A correlation between the type of herd and the presence of M hyopneumoniae was statistically insignificant and no effect of farrowing rhythm could be confirmed.

Livestock associated methicillin-resistant Staphylococcus aureus (LaMRSA) isolated from lesions of pigs at necropsy in northwest Germany between 2004 and 2007.

An increasing number of reported detections of methicillin-resistant Staphylococcus aureus (MRSA) in food animals since 2007 has led to the assumption that there is an emerging zoonotic problem with livestock associated (la)MRSA potentially aggravating the MRSA problem in humans. It was the objective of the study to investigate, whether MRSA was present in clinical specimens of pigs collected at post-mortem since 2004 and to further characterize these isolates. We studied 138 isolates of S. aureus collected between 2004 and 2007 from various pathological lesions of pigs at necropsy. Potential MRSA were identified by growth on selective chromogenic media. Isolates were confirmed as MRSA using multiplex PCR. Confirmed isolates were spa- and SCCmec-typed and were tested for antimicrobial resistance. Overall, 60 (43%) S. aureus isolates were identified as MRSA. The majority (57/60) of the MRSA isolates found in the altered porcine tissues were spa-types associated with MRSA ST398. Three MRSA were ST97 isolates, a type that has not been described as an MRSA in pigs before. Other clonal complexes (ST9, ST30) dominated among the methicillin-sensitive S. aureus. MRSA were found in similar frequency in all 4 years. We assume that MRSA in pigs may have occurred earlier than 2004 and might be not really 'emerging', but rather have been overlooked until recently. The potentially causative role of the MRSA in the lesions warrants further investigation.

[Bronchoalveolar lavage on pig breeding farms]. / Lungenspülungen bei Schweinen im Erzeugerbetrieb.

In 182 pigs lung lavage was performed using a endotracheal tube and a catheter. The collected bronchoalveolar lavage fluid (BALF) was examined microbiologically. With decreasing numbers alpha-hämolytic Streptococci, Bordetella bronchiseptica, Haemophilus parasuis, Pasteurella multocida were cultured. Actinobacillus pleuropneumoniae was isolated from 3 BALFs. In one farm piglets were lavaged routinely for monitoring of the lung health status.

The 13C-urea breath test--detection of Helicobacter pylori infection in patients with partial gastrectomy.

The 13C-urea breath test (13C-UBT) for diagnosis of Helicobacter pylori (Hp) infection was evaluated in 41 patients after partial gastrectomy and was used for determination of the Hp-prevalence after two different procedures of reconstruction of the gastrointestinal tract, i.e. Billroth's II operation and Roux-en-Y anastomosis. Breath samples were taken at various time points within 30 minutes after a motility inhibiting liquid test meal with citric acid followed by 75 mg of 13C-urea. The 13CO2/12CO2-ratio (delta-value) was measured using isotope ratio mass spectrometry and the recovery of tracer in the exhaled breath was calculated (UBT-value). In all patients and in the corresponding control groups comparison of established reference methods (culture, CLO test, and Fuchsin staining) with the 4 point breath analysis for detection of Hp was investigated. In patients with partial gastrectomy, the sensitivity of the 13C-UBT to detect the presence of Hp and the negative predictive values were 100%, whereas the specificity and the positive predictive values were about 80%. In patients without gastric surgery quality control parameters were not significantly different. Hp-prevalence in postoperative patients was about 45%. All results were independent of their expression either as delta-value or as UBT-value and were not significantly different between the patients with Billroth's II operation and the patients with Roux-en-Y anastomosis. In conclusion, the 13C-UBT is a suitable method for diagnosis and therapeutic monitoring of Hp-status in patients after partial gastrectomy.

The simplified 13C-urea breath test--one point analysis for detection of Helicobacter pylori infection.

A 13C-urea breath test for detection of Helicobacter pylori infection was validated in 306 patients. Breath samples (four, two or one) were taken at various time points within 30 minutes after a motility inhibiting liquid test meal with citric acid followed by 75 mg of 13C-urea. The 13CO2/12 CO2-ratio (delta-value) was measured using isotope ratio mass spectrometry and the recovery of tracer in the exhaled breath was calculated (UBT-value). In 172 patients a comparison of established reference methods (culture, CLO test, and Fuchsin staining) with the 4 point breath analysis for detection of Helicobacter pylori showed high values of quality parameters (greater than 80% for sensitivity, specificity, positive and negative predictive value), independent of expression of the results as delta-value or as UBT-value. In 134 patients a reduction of samples to one single breath sample taken 30 minutes after ingestion of the tracer showed no significant differences in the quality parameters when compared with the standard 4 point breath analysis. This indicates that the analysis of a single breath sample is suitable for detection of Helicobacter pylori status in man.

[Helicobacter pylori colonization in surgical patients]. / Untersuchungen zur Helicobacter-pylori-Besiedlung bei operierten Patienten.

The prevalence of Helicobacter pylori (HP) was examined in 387 patients undergoing endoscopy of the upper gastrointestinal tract. Of central interest was the question to which extent surgical intervention influences the colonisation of the gastric mucosa with HP. The bacillic status was appraised using double microbiological examinations, histological determination and the CLO-test. In 229 patients a 13C-urea-breath test was also carried out (sensitivity 98%). HP could be detected in 90% of all patients presenting with duodenal ulcers as well as in 70% of patients with gastric ulcers, whereas in those patients in whom a lesion of the upper gastrointestinal tract could be excluded through endoscopy. HP was found in only 27%. The prevalence of HP did not increase with age. In patients having undergone distal gastric resection due to gastric ulcers, HP was only rarely found (19%) in the mucosa in the vicinity of the anastomosis following removal of the apparently pathogenetically important antrum mucosa. There was no association between anastomosis ulcers and bacillic colonisation. Following selective proximal vagotomy in patients with duodenal ulcers, HP was found in 80% of all cases. In these patients there was also no association between recurrent ulceration and a HP-positive status. Our results describe the postoperative HP-status after different surgical procedures of ulcer therapy: whereas a distal gastric resection removes the antrum mucosa, which provides the necessary environmental milieu, the HP-colonisation rate after selective proximal vagotomy is similar to that in non-operated ulcer disease.

A comparison of secretory proteinases from different strains of Candida albicans.

Randomly selected strains of Candida albicans were grown with bovine serum albumin (BSA) as a single nitrogen source. From all strains tested, culture supernatant contained carboxyl proteinase (E.C.3.4.23) as has been shown that with hemoglobin as a substrate and by specific inhibition with pepstatin-A. According to the separation pattern of BSA fragments, secretory proteinases from C. albicans belong to at least three groups. We have purified the partially proteolytic enzyme of strain 113 and have compared its properties with those of the totally proteolytic enzyme of strain CBS 2730. Both enzymes have virtually identical molecular weight (ca. 44,000) and cross-react immunologically; they differ in pH optimum, isoelectric point, substrate specificity, and resistance against alkali. IgG1, which is the prevalent immunoglobulin of human serum, was not cleaved by enzyme 113. Immunoglobulins A1, A2 and secretory component were cleaved by both enzymes, which points to a role of the secretory proteinases in the persistence of yeasts on mucous membranes. Differences in the course of alkaline denaturation indicate that only a fraction of strain-specific proteinases is capable to convey long-range effects in the host.