Genetic basis of penicillin resistance of S. aureus isolated in bovine mastitis

Background

The blaZ gene encoding penicillin resistance can be located either chromosomally or on plasmids. The aim of this study was to investigate the genetic relationships and to determine the location of the blaZ gene in S. aureus isolated in bovine mastitis in Finland and Sweden.

Methods

Seventy-eight β-lactamase positive S. aureus isolates from bovine mastitis (34 from Finland and 44 from Sweden) were included in the study. The localization of blaZ gene was determined by Southern blotting. The blaZ genes of the isolates were sequenced and the sequences were translated to beta-lactamase proteins and further grouped as different protein signatures. The isolates and, as control, 33 Swedish and 36 Finnish beta-lactamase negative isolates were typed with pulsed-field gel electrophoresis (PFGE).

Results

In 26 out of 34 Finnish isolates (76.5%) and in 25 out of 44 Swedish isolates (56.8%) the blaZ gene was localized on a plasmid. Six different protein signatures were found. One signature was found only in four Swedish isolates, but all other signatures were found both in Finnish and Swedish isolates. The PFGE results revealed a diversity of S. aureus clones. The protein signatures were not clearly associated with certain pulsotypes.

Conclusions

The plasmid location of the blaZ gene was not statistically significantly more common in Finland than in Sweden, and hence does not explain the higher proportion of penicillin-resistant isolates of S. aureus causing bovine mastitis in Finland compared to Sweden.     

Characterization of In Vitro Synthesized Equine Metabolites of the Selective Androgen Receptor Modulators S24 and S4

Several selective androgen receptor modulators (SARMs) have been synthesized and investigated in humans, rats, and dogs in the past, but no data are yet available concerning the metabolism of SARMs in horses. The aryl-propionamide-derived drug candidates S24 and S4 (andarine) have a strong androgen receptor binding affinity and show distinctive specific cell answers. Although no SARM drug candidate (aiming for testosterone replacement therapy) has completed clinical trials yet, S4 has been illicitly available via the Internet. These facts led to the prohibition of SARMs by the German equestrian federation, and the (mis)use of such compounds would further represent a doping rule violation in horse racing. In this study, the drug candidates S24 and S4 were subjected to in vitro metabolism experiments with equine liver microsomal preparations from a female Quarter Horse to obtain information about potential target analytes in equine doping control analysis. The enzymatically synthesized metabolites were characterized by liquid chromatography–tandem mass spectrometry and –high-resolution/high-accuracy mass spectrometry. All observed S24 and S4 equine metabolites are in agreement with earlier in vitro and in vivo studies in humans and dogs. Nevertheless, the relative percentage of generated equine metabolites (as determined from the analytes’ response in full-scan chromatography–tandem mass spectrometry and –high-resolution/high-accuracy mass spectrometry measurements) differs considerably from the reported profiles. Although the S24 metabolite pattern is comparably balanced concerning glucuronidated and sulfonated conjugates, the major S4 metabolite was found to be the unconjugated dephenylated compound, with a proportion of more than 90%.     

Modeling carbon allocation in trees: a search for principles

We review approaches to predicting carbon and nitrogen allocation in forest models in terms of their underlying assumptions and their resulting strengths and limitations. Empirical and allometric methods are easily developed and computationally efficient, but lack the power of evolution-based approaches to explain and predict multifaceted effects of environmental variability and climate change. In evolution-based methods, allocation is usually determined by maximization of a fitness proxy, either in a fixed environment, which we call optimal response (OR) models, or including the feedback of an individual's strategy on its environment (game-theoretical optimization, GTO). Optimal response models can predict allocation in single trees and stands when there is significant competition only for one resource. Game-theoretical optimization can be used to account for additional dimensions of competition, e.g., when strong root competition boosts root allocation at the expense of wood production. However, we demonstrate that an OR model predicts similar allocation to a GTO model under the root-competitive conditions reported in free-air carbon dioxide enrichment (FACE) experiments. The most evolutionarily realistic approach is adaptive dynamics (AD) where the allocation strategy arises from eco-evolutionary dynamics of populations instead of a fitness proxy. We also discuss emerging entropy-based approaches that offer an alternative thermodynamic perspective on allocation, in which fitness proxies are replaced by entropy or entropy production. To help develop allocation models further, the value of wide-ranging datasets, such as FLUXNET, could be greatly enhanced by ancillary measurements of driving variables, such as water and soil nitrogen availability.     

The association between rainfall and seropositivity to Leptospira in outdoor reared pigs

Outdoor reared pigs were used as indicators for investigating the effect of weather conditions in the seroprevalence of Leptospira. Over the period February to March 2008, sera from 386 sows on 11 farms in southern Sweden were tested for antibodies to the following Leptospira serovars: L. interrogans serovar (sv) Bratislava, L. kirschneri sv Grippotyphosa, L. interrogans sv Icterohaemorrhagiae, L. interrogans sv Pomona, L. borgpetersenii sv Tarassovi and one domestic strain (mouse 2A) related to L. borgpetersenii sv Sejroe and L. borgpetersenii sv Istrica. The highest seroprevalence was to this strain (8.0%) followed by sv Bratislava (3.9%). Six of the 11 farms had sows which were seropositive to at least one of the Leptospira serovars. Data on rainfall and temperature were retrieved for the respective farms. For each millimetre of extra rainfall, there was an increase in the odds ratio (OR) for seropositivity to sv Bratislava of 4.3 (95% CI 1.9-10), and to strain mouse 2A of 2.5 (95% CI 1.0-6.4). There was no association between seropositivity and temperature. This study indicates that different climate conditions within the northern temperate climate zone may be of importance for the presence of Leptospira-seropositivity in mammals.     

Genetic and Environmental Correlations Among Female Fertility Traits and Milk Production in Different Parities of Swedish Red and White Dairy Cattle

The main objective of this study was to estimate genetic correlations between fertility and production traits in first, second and third lactations as well as between fertility traits measured in the same way at different ages. The fertility traits studied were: number of inseminations per service period, number of treatments for reproductive disturbances, interval between first and last inseminations, interval between calving and first insemination, and interval between calving and last insemination. Early milk production was measured as the average of the energy-corrected milk yield at the second and third monthly testdays in a lactation. The number of records was approximately 450 000, 350 000, 180 000 and 75 000 in the heifer period, first, second, and third lactations, respectively. A linear, trivariate model that included the effects of herd-year, year, month, age and sire of the cow was applied. To reduce the effect of ongoing selection, 305-days kg protein production in first lactation was included as a variate in all of the analyses. Correlations between the herd-year effects indicated that factors of herd-year level conducive to increased production had a tendency to increase the number of inseminations as well as the number of reproductive treatments, although there was an earlier start and termination of the insemination period. Genetic correlations between fertility traits and production were in the range of 0.2-0.4, all of them unfavourable and higher at later parities. The genetic correlations between fertility traits in the heifer period and the same traits in first lactation were 0.7. Genetic correlations between the first and second lactation varied between 0.7 and 0.9, and between the second and third lactation they were all 0.9 or higher. In conclusion, fertility and production traits need to be selected for simultaneously if fertility is going to be maintained along further genetic improvement on production, and such selection should include fertility results from lactating cows.     

Cross-sectional study on acrylamide hemoglobin adducts in subpopulations from the European Prospective Investigation into Cancer and Nutrition (EPIC) Study

Acrylamide exposure was investigated in subgroups of the EPIC study population (510 subjects from 9 European countries, randomly selected and stratified by age, gender, and smoking status) using hemoglobin adducts of acrylamide (HbAA) and its primary metabolite glycidamide (HbGA). Blood samples were analyzed for HbAA and HbGA by HPLC/MS/MS. Statistical models for HbAA and HbGA were developed including body mass index (BMI), educational level, and physical activity. A large variability in acrylamide exposure and metabolism between individuals and country groups was observed with HbAA and HbGA values ranging between 15-623 and 8-377 pmol/g of Hb, respectively. Both adducts differed significantly by country, sex, and smoking status. HbGA values were significantly lower in high alcohol consumers than in moderate consumers. With increasing BMI, HbGA in nonsmokers and HbAA in smokers decreased significantly. In the assessment of potential health effects related to acrylamide exposure, country of origin, BMI, alcohol consumption, sex, and smoking status should be considered.     

Evaluation of a novel proximity ligation assay for the sensitive and rapid detection of foot-and-mouth disease virus

A novel proximity ligation assay (PLA) using a pan-serotype reactive monoclonal antibody was developed and evaluated for the detection of foot-and-mouth disease virus (FMDV) in clinical samples collected from field cases of disease. The FMDV-specific PLA was found to be 100 times more sensitive for virus detection than the commonly used antigen capture-ELISA (AgELISA). As few as five TCID50 were detected in individual assays, which was comparable with the analytical sensitivity of real-time RT-PCR. Although this assay was capable of detecting diverse isolates from all seven FMDV serotypes, the diagnostic sensitivity of the PLA assay was lower than real-time RT-PCR mainly due to a failure to detect some SAT 1, SAT 2 and SAT 3 FMDV strains. In conclusion, this new PLA format has high analytical sensitivity for the detection of FMDV in clinical samples and may prove valuable as a rapid and simple tool for use in FMD diagnosis.     

Metabolic profiles in five high-producing Swedish dairy herds with a history of abomasal displacement and ketosis

Background

Body condition score and blood profiles have been used to monitor management and herd health in dairy cows. The aim of this study was to examine BCS and extended metabolic profiles, reflecting both energy metabolism and liver status around calving in high-producing herds with a high incidence of abomasal displacement and ketosis and to evaluate if such profiles can be used at herd level to pinpoint specific herd problems.

Methods

Body condition score and metabolic profiles around calving in five high-producing herds with high incidences of abomasal displacement and ketosis were assessed using linear mixed models (94 cows, 326 examinations). Cows were examined and blood sampled every three weeks from four weeks ante partum (ap) to nine weeks postpartum (pp). Blood parameters studied were glucose, fructosamine, non-esterified fatty acids (NEFA), insulin, β-hydroxybutyrate, aspartate aminotransferase, glutamate dehydrogenase, haptoglobin and cholesterol.

Results

All herds had overconditioned dry cows that lost body condition substantially the first 4–6 weeks pp. Two herds had elevated levels of NEFA ap and three herds had elevated levels pp. One herd had low levels of insulin ap and low levels of cholesterol pp. Haptoglobin was detected pp in all herds and its usefulness is discussed.

Conclusion

NEFA was the parameter that most closely reflected the body condition losses while these losses were not seen in glucose and fructosamine levels. Insulin and cholesterol were potentially useful in herd profiles but need further investigation. Increased glutamate dehydrogenase suggested liver cell damage in all herds.     

Therapeutic effects of systemic or intramammary antimicrobial treatment of bovine subclinical mastitis during lactation

The short- and long-term treatment efficacy of administrating penicillin for bovine subclinical mastitis during lactation when using intramuscular (IM; 9.5 mg [15,000 IU]/kg bodyweight of benzyl penicillin potassium) injections twice daily for 5 days, or intramammary (IMM; 0.3g [300,000 IU] penethamate hydroiodide) administration once daily for 5 days was compared with a control group receiving no treatment. One hundred and twenty-six cows met the inclusion criteria, which were lack of clinical symptoms, no recent treatment with antimicrobials, and findings of penicillin-sensitive Staphylococcus aureus, Streptococcus dysgalactiae, or Streptococcus uberis in combination with an inflammatory reaction. At follow-up 42-58 days after treatment, the proportion of cows negative for the original infection was significantly higher in IM and IMM groups compared to controls, but the difference between antimicrobial treatment groups was not significant. The udder quarter milk somatic cell count (SCC) was significantly lower at follow-up in IM and IMM groups than in controls, but milk production did not differ between treatments. The culling rate during the 10-month period following treatment was significantly higher in the group treated with IMM penicillin than in the other two groups, but the risk of new mastitis treatments within 10 months did not differ between the three groups. The cure rate was significantly affected by lactation number (lower in older cows), breed (lower in the Swedish Holstein breed), pathogen (lower for S. aureus), and pre-treatment SCC (higher for above average SCC). In conclusion, beneficial long-term effects of antimicrobial treatment during lactation of subclinical mastitis caused by S. aureus, Str. dysgalactiae or Str. uberis were not found in the present study.