Engineering a glycerol utilization pathway in Corynebacterium glutamicum for succinate production under O<Subscript>2</Subscript> deprivation

Objective

To explore the glycerol utilization pathway in Corynebacterium glutamicum for succinate production under O₂ deprivation.

Result

Overexpression of a glycerol facilitator, glycerol dehydrogenase and dihydroxyacetone kinase from Escherichia coli K-12 in C. glutamicum led to recombinant strains NC-3G diverting glycerol utilization towards succinate production under O₂ deprivation. Under these conditions, strain NC-3G efficiently consumed glycerol and produced succinate without growth. The recombinant C. glutamicum utilizing glycerol as the sole carbon source showed higher intracellular NADH/NAD⁺ ratio compare with utilizing glucose. The mass conversion of succinate increased from 0.64 to 0.95. Using an anaerobic fed-batch fermentation process, the final strain produced 38.4 g succinate/l with an average yield of 1.02 g/g.

Conclusions

The metabolically-engineered strains showed an efficient succinate production using glycerol as sole carbon source under O₂ deprivation.

     

Co-cultivation of <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis> with <Emphasis Type="Italic">Azotobacter chroococcum</Emphasis> improved H<Subscript>2</Subscript> production

Objectives

To improve H₂ production, the green algae Chlamydomonas reinhardtii cc849 was co-cultured with Azotobacter chroococcum.

Results

The maximum H₂ production of the co-culture was 350% greater than that of the pure algal cultures under optimal H₂ production conditions. The maximum growth and the respiratory rate of the co-cultures were about 320 and 300% of the controls, and the dissolved O₂ of co-cultures was decreased 74%. Furthermore, the in vitro maximum hydrogenase activity of the co-culture was 250% greater than that of the control, and the in vivo maximum hydrogenase activity of the co-culture was 1.4-fold greater than that of the control. In addition, the maximum starch content of co-culture was 1400% that of the control.

Conclusions

Azotobacter chroococcum improved the H₂ production of the co-cultures by decreasing the O₂ content and increasing the growth and starch content of the algae and the hydrogenase activity of the co-cultures relative to those of pure algal cultures.

     

Biomanufacture of nano-Pd(0) by <Emphasis Type="Italic">Escherichia coli</Emphasis> and electrochemical activity of bio-Pd(0) made at the expense of H<Subscript>2</Subscript> and formate as electron donors

Objective

Palladised cells of Desulfovibrio desulfuricans and Shewanella oneidensis have been reported as fuel cell electrocatalysts but growth at scale may be unattractive/costly; we have evaluated the potential of using E. coli, using H₂/formate for Pd-nanoparticle manufacture.

Results

Using ‘bio-Pd’ made under H₂ (20 wt%) cyclic voltammograms suggested electrochemical activity of bio-NPs in a native state, attributed to proton adsorption/desorption. Bio-Pd prepared using formate as the electron donor gave smaller, well separated NPs; this material showed no electrochemical properties, and hence little potential for fuel cell use using a simple preparation technique. Bio-Pd on S. oneidensis gave similar results to those obtained using E. coli.

Conclusion

Bio-Pd is sufficiently conductive to make an E. coli-derived electrochemically active material on intact, unprocessed bacterial cells if prepared at the expense of H₂, showing potential for fuel cell applications using a simple one-step preparation method.

     

The effect of 1,25-dihydroxyvitamin D<Subscript>3</Subscript> on TSLP,IL-33 and IL-25 expression in respiratory epithelium

Background

Airway epithelium is an active and important component of the immunological response in the pathophysiology of obstructive lung diseases. Recent studies suggest an important role for vitamin D₃ in asthma severity and treatment response.

Objective

Our study evaluated the influence of an active form of vitamin D₃ on the expression of selected mediators of allergic inflammation in the respiratory epithelium.

Material and Methods

Primary nasal and bronchial epithelial cells were exposed to1,25D₃ for 1 hour and were then stimulated or not with IL-4, TNF-α, LPS, and poly I:C. After 24 hours TSLP, IL-33, and IL-25 protein levels were measured in culture supernatants usingELISAandmRNAlevels in cells by real time PCR.

Results

1,25D₃ increased TSLP concentration in unstimulated nasal epithelial cells, but did not influence IL-33 and IL-25 expression. In IL-4-stimulated epithelial cell cultures 1,25D₃ mostly inhibited TSLP and IL-33 expression. In LPS-treated cultures 1,25D₃ decreased IL-33 expression. Simultaneously 1,25D₃ augmented IL-25 production in the same model of stimulation.

Conclusion

Our study revealed the dual nature of vitamin D₃ manifested in both pro- and anti-inflammatory properties observed in airway epithelial cells.

     

Transmission and characterization of <Emphasis Type="Italic">bla</Emphasis><Subscript>NDM-1</Subscript> in <Emphasis Type="Italic">Enterobacter cloacae</Emphasis> at a teaching hospital in Yunnan,China

Background

In recent years, New Delhi metallo-beta-lactamases 1 (bla _NDM-1) has been reported with increasing frequency and become prevalent. The present study was undertaken to investigate the epidemiological dissemination of the bla _NDM-1 gene in Enterobacter cloacae isolates at a teaching hospital in Yunnan, China.

Methods

Antimicrobial susceptibility testing was performed using VITEK 2 system and E test gradient strips. The presence of integrons and insertion sequence common region 1 were examined by PCR and sequencing. Clonal relatedness was assessed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing. Conjugation experiments and Southern blot hybridization were performed to determine the transferability of plasmids.

Results

Ten E. cloacae isolates and their Escherichia coli transconjugants were exhibited similar resistant patterns to carbapenems, cephalosporins and penicillins. 8 (80%) of E. cloacae isolates carried class 1 integron and 1 (12.5%) carried class 2 integron. Integron variable regions harbored the genes which encoded resistance to aminoglycosides (aadA1, aadA2, aadA5, aadB, aac(6′)-Ib-cr), sulfamethoxazole/trimethoprim (dfrA17, dfrA12, dfrA15) and Streptozotocin (sat2). Six E. cloacae isolates belonged to ST74 and exhibited highly similar PFGE patterns. Each isolate shared an identical plasmid with ~33.3 kb size that carried the bla _NDM-1 gene, except T3 strain, of which the bla _NDM-1 gene was located on a ~50 kb plasmid.

Conclusions

Our findings suggested that plasmid was able to contribute to the dissemination of bla _NDM-1. Hence, more attention should be devoted to monitor the dissemination of the bla _NDM-1 gene due to its horizontal transfer via plasmid. In addition, nosocomial surveillance system should actively monitor the potential endemic clone of ST74 to prevent their further spread.

     

Microbial community and associated enzymes activity influence soil carbon chemical composition in <Emphasis Type="Italic">Eucalyptus urophylla</Emphasis> plantation with mixing N<Subscript>2</Subscript>-fixing species in subtropical China

Background and aims

Microbial communities and their associated enzyme activities affect the quantity and chemical quality of carbon in soil. We aimed to evaluate the biochemical mechanisms underlying how N₂-fixing species influences soil organic carbon chemical composition through soil microbial functional groups and enzyme activities.

Methods

We examined the effects of N₂-fixing species mixed with Eucalyptus on soil carbon storage, and the chemical composition of an 8-year-old pure Eucalyptus urophylla plantation (PP) and a mixed E.urophylla and Acacia mangium plantation (MP).

Results

The soil carbon stock and recalcitrant carbon chemical component significantly increased in surface soil in MP. The total PLFAs and bacterial PLFAs increased by 29.1% and 27.0% in cool-dry season, while in the warm-wet season, the total PLFAs and bacterial PLFAs increased by 13.1% and 27.3%, respectively. However, the fungal PLFAs decreased significantly in warm-wet season in MP. The total activity of the cellulose-degrading enzyme β-glucosidase was significantly greater with mixed N₂-fixing species in both dry-cool and wet-warm season. The increase in the Alk-C/O-Alk-C ratio and SOC was strongly associated with both C-acquisition activity and bacterial community.

Conclusions

Our findings highlight the importance of N₂-fixing species in regulating both soil microbial communities and their functioning in association with soil extracellular enzyme activities, which contribute to the increased soil carbon storage and recalcitrant carbon composition in Eucalyptus plantations.

     

Iridoid and phenylethanoid/phenylpropanoid metabolite profiles of <Emphasis Type="Italic">Scrophularia</Emphasis> and <Emphasis Type="Italic">Verbascum</Emphasis> species used medicinally in North America

Introduction

Botanicals containing iridoid and phenylethanoid/phenylpropanoid glycosides are used worldwide for the treatment of inflammatory musculoskeletal conditions that are primary causes of human years lived with disability, such as arthritis and lower back pain.

Objectives

We report the analysis of candidate anti-inflammatory metabolites of several endemic Scrophularia species and Verbascum thapsus used medicinally by peoples of North America.

Methods

Leaves, stems, and roots were analyzed by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) and partial least squares-discriminant analysis (PLS-DA) was performed in MetaboAnalyst 3.0 after processing the datasets in Progenesis QI.

Results

Comparison of the datasets revealed significant and differential accumulation of iridoid and phenylethanoid/phenylpropanoid glycosides in the tissues of the endemic Scrophularia species and Verbascum thapsus.

Conclusions

Our investigation identified several species of pharmacological interest as good sources for harpagoside and other important anti-inflammatory metabolites.

     

Long-term H<Subscript>2</Subscript> photoproduction from starch by co-culture of <Emphasis Type="Italic">Clostridium butyricum</Emphasis> and <Emphasis Type="Italic">Rhodobacter sphaeroides</Emphasis> in a repeated batch process

Objectives

To prove the possibility of efficient starch photofermentation in co-culture of heterotrophic and phototrophic bacteria over prolonged period.

Results

Repeated batch photofermentation of starch was demonstrated in co-culture Clostridium butyricum and Rhodobacter sphaeroides under microaerobic conditions. It continued 15 months without addition of new inoculum or pH regulation when using 4–5 g starch l^?1 and 0.04 g yeast extract l^?1. The complete degradation of starch without volatile fatty acids accumulation was shown in this co-culture. The average H₂ yield of 5.2 mol/mol glucose was much higher than that in Clostridium monoculture. The species composition of co-culture was studied by q-PCR assay. The concentration of Clostridium cells in prolonged co-culture was lower than in monoculture and even in a single batch co-culture. This means that Clostridia growth was significantly limited whereas starch hydrolysis still took place.

Conclusion

The prolonged repeated batch photofermentation of starch by co-culture C. butyricum and R. sphaeroides provided efficient H₂ production without accumulation of organic acids under conditions of Clostridia limitation.

     

Pathway-specific metabolome analysis with <Superscript>18</Superscript>O<Subscript>2</Subscript>-labeled <Emphasis Type="Italic">Medicago truncatula</Emphasis> via a mass spectrometry-based approach

Introduction

Oxygen from carbon dioxide, water or molecular oxygen, depending on the responsible enzyme, can lead to a large variety of metabolites through chemical modification.

Objectives

Pathway-specific labeling using isotopic molecular oxygen (¹⁸O₂) makes it possible to determine the origin of oxygen atoms in metabolites and the presence of biosynthetic enzymes (e.g., oxygenases). In this study, we established the basis of ¹⁸O₂-metabolome analysis.

Methods

¹⁸O₂ labeled whole Medicago truncatula seedlings were prepared using ¹⁸O₂-air and an economical sealed-glass bottle system. Metabolites were analyzed using high-accuracy and high-resolution mass spectrometry. Identification of the metabolite was confirmed by NMR following UHPLC–solid-phase extraction (SPE).

Results

A total of 511 peaks labeled by ¹⁸O₂ from shoot and 343 peaks from root were annotated by untargeted metabolome analysis. Additionally, we identified a new flavonoid, apigenin 4′-O-[2′-O-coumaroyl-glucuronopyranosyl-(1–2)-O-glucuronopyranoside], that was labeled by ¹⁸O₂. To the best of our knowledge, this is the first report of apigenin 4′-glucuronide in M. truncatula. Using MSⁿ analysis, we estimated that ¹⁸O atoms were specifically incorporated in apigenin, the coumaroyl group, and glucuronic acid. For apigenin, an ¹⁸O atom was incorporated in the 4′-hydroxy group. Thus, non-specific incorporation of an ¹⁸O atom by recycling during one month of labeling is unlikely compared with the more specific oxygenase-catalyzing reaction.

Conclusion

Our finding indicated that ¹⁸O₂ labeling was effective not only for the mining of unknown metabolites which were biosynthesized by oxygenase-related pathway but also for the identification of metabolites whose oxygen atoms were derived from oxygenase activity.

     

<Emphasis Type="Italic">Artemisia frigida</Emphasis> and <Emphasis Type="Italic">Stipa krylovii</Emphasis>, two dominant species in Inner Mongolia steppe,differed in their responses to elevated atmospheric CO<Subscript>2</Subscript> concentration

Aims

Despite extensive studies on effects of elevated CO₂ concentration ([CO₂]_e) on plant growth, few studies have investigated the responses of native grassland plant species to [CO₂]_e in terms of nutrient acquisition.

Methods

The effects of [CO₂]_e (769 ± 23 ppm) on Artemisia frigida and Stipa krylovii, two dominant species in Inner Mongolia steppe were investigated by growing them for 7 weeks in Open-Top Chambers (OTC).

Results

Exposure to [CO₂]_e enhanced shoot and root growth of A. frigida and S. krylovii. Elevated [CO₂] increased photosynthetic rates (Pn) by 34 % in A. frigida but decreased Pn by 52 % in S. krylovii. Moreover, root-secreted acid phosphatase activity in A. frigida was stimulated by [CO₂]_e, while exudation of malate from roots of S. krylovii was suppressed by [CO₂]_e. Exposure to [CO₂]_e led to a decrease in P concentration in shoots and roots of A. frigida and S. krylovii, but total amount of P accumulated in shoots and roots of both species was increased by [CO₂]_e.

Conclusions

The two dominant species in temperate steppes differed in their responses to [CO₂]_e, such that A. frigida was more adapted to [CO₂]_e than S. krylovii under low availability of soil P.

     

Dual stimuli-responsive Fe<Subscript>3</Subscript>O<Subscript>4</Subscript> graft poly(acrylic acid)-<Emphasis Type="Italic">block</Emphasis>-poly(2-methacryloyloxyethyl ferrocenecarboxylate) copolymer micromicelles: surface RAFT synthesis,self-assembly and drug release applications

Background

Stimuli-responsive polymer materials are a new kind of intelligent materials based on the concept of bionics, which exhibits more significant changes in physicochemical properties upon triggered by tiny environment stimuli, hence providing a good carrier platform for antitumor drug delivery.

Results

Dual stimuli-responsive Fe₃O₄ graft poly(acrylic acid)-block-poly(2-methacryloyloxyethyl ferrocenecarboxylate) block copolymers (Fe₃O₄-g-PAA-b-PMAEFC) were engineered and synthesized through a two-step sequential reversible addition-fragmentation chain transfer polymerization route. The characterization was performed by FTIR, ¹H NMR, SEC, XRD and TGA techniques. The self-assembly behavior in aqueous solution upon triggered by pH, magnetic and redox stimuli was investigated via zeta potentials, vibration sample magnetometer, cyclic voltammetry, fluorescent spectrometry, dynamic light scattering, XPS, TEM and SEM measurements. The experimental results indicated that the Fe₃O₄-g-PAA-b-PMAEFC copolymer materials could spontaneously assemble into hybrid magnetic copolymer micromicelles with core–shell structure, and exhibited superparamagnetism, redox and pH stimuli-responsive features. The hybrid copolymer micromicelles were stable and nontoxic, and could entrap hydrophobic anticancer drug, which was in turn swiftly and effectively delivered from the drug-loaded micromicelles at special microenvironments such as acidic pH and high reactive oxygen species.

Conclusion

This class of stimuli-responsive copolymer materials is expected to find wide applications in medical science and biology, etc., especially in drug delivery system.

     

Determination of Azole Resistance and TR<Subscript>34</Subscript>/L98H Mutations in Isolates of <Emphasis Type="Italic">Aspergillus</Emphasis> Section <Emphasis Type="Italic">Fumigati</Emphasis> from Turkish Cystic Fibrosis Patients

Background

Aspergillus fumigatus is the species section Fumigati most frequently isolated from the respiratory tract of cystic fibrosis (CF) patients. Recent studies suggest that mutations in the Cyp51 gene, particularly TR₃₄/L98H, are responsible for azole resistance.

Objectives and Methods

The focus of this study was on section Fumigati isolates isolated from the respiratory tract samples of CF patients. More specifically, the goal was to detect A. fumigatus isolates, test their antifungal susceptibility to itraconazole, voriconazole and posaconazole, and finally determine the presence of TR₃₄/L98H and other mutations in the isolates Cyp51A gene.

Results and Conclusions

A set of 31 isolates of Aspergillus section Fumigati were obtained from the sputum samples of 6 CF patients and subsequently identified to species level by microsatellite genotyping. All isolates were determined as A. fumigatus and involved 14 different genotypes. The minimal inhibitory concentrations to the three azoles were determined by the E-test method, and the Cyp51A gene was sequenced. One of the genotypes was found to be resistant to all azoles but no mutations were detected in the Cyp51A gene, especially the TR₃₄/L98H mutation. Therefore, mutations in genes other than Cyp51A or other distinct mechanisms may be responsible for this reported multiazole resistance found in a Turkish CF patient.

     

Effectiveness of fractional CO<Subscript>2</Subscript> laser in women with stress urinary incontinence

Background

Stress urinary incontinence (SUI) is a relatively common disorder that significantly affects the quality of life. Many conservative and surgical treatment methods have been recommended for SUI, but they have major limitations.

Aims

To assess the use of the CO₂ fractional laser in the treatment of SUI.

Methods

This clinical trial included 55 patients with confirmed SUI. Patients underwent fractional CO₂ laser treatment 3 times at 30-day intervals. Data on age, smoking history, sexual activity, menopause, and history of hormone replacement therapy (HRT) were collected. Response to treatment was assessed by SUI severity and the level of sexual satisfaction was assessed using the visual analog scale (VAS). Patients were evaluated at 3 different time points: before treatment, and 45 days and 6 months after the last laser treatment.

Results

The mean patient age was 44.4±11.4 years (range: 28 to 68 years). Smoking history was positive in 6 patients (9.1%); 19 (54.3%) were menopausal on HRT. The SUI severity score at baseline (before treatment) was 8.56±0.62 and decreased to 2.28 6 months after treatment (p<0.0001). The sexual satisfaction score was 3±0.94 at baseline and increased to 7.87±0.93 6 months after treatment (day 180) (p<0.0001, slope = + 2.2)

Conclusion

Our findings are in line with a previous study that showed the value of transvaginal CO₂ fractional laser treatment for alleviation of SUI symptoms and its potential as an alternative treatment. We also observed improved sexual satisfaction in SUI patients.

     

Contribution of glutamate decarboxylase in <Emphasis Type="Italic">Lactobacillus reuteri</Emphasis> to acid resistance and persistence in sourdough fermentation

Background

Acid stress impacts the persistence of lactobacilli in industrial sourdough fermentations, and in intestinal ecosystems. However, the contribution of glutamate to acid resistance in lactobacilli has not been demonstrated experimentally, and evidence for the contribution of acid resistance to the competitiveness of lactobacilli in sourdough is lacking. It was therefore the aim of this study to investigate the ecological role of glutamate decarboxylase in L. reuteri.

Results

A gene coding for a putative glutamate decarboxylase, gadB, was identified in the genome of L. reuteri 100-23. Different from the organization of genetic loci coding for glutamate decarboxylase in other lactic acid bacteria, gadB was located adjacent to a putative glutaminase gene, gls3. An isogenic deletion mutant, L. reuteri ?gadB, was generated by a double crossover method. L. reuteri 100-23 but not L. reuteri ?gadB converted glutamate to γ-aminobutyrate (GABA) in phosphate butter (pH 2.5). In sourdough, both strains converted glutamine to glutamate but only L. reuteri 100-23 accumulated GABA. Glutamate addition to phosphate buffer, pH 2.5, improved survival of L. reuteri 100-23 100-fold. However, survival of L. reuteri ?gadB remained essentially unchanged. The disruption of gadB did not affect growth of L. reuteri in mMRS or in sourdough. However, the wild type strain L. reuteri 100-23 displaced L. reuteri ?gadB after 5 cycles of fermentation in back-slopped sourdough fermentations.

Conclusions

The conversion of glutamate to GABA by L. reuteri 100-23 contributes to acid resistance and to competitiveness in industrial sourdough fermentations. The organization of the gene cluster for glutamate conversion, and the availability of amino acids in cereals imply that glutamine rather than glutamate functions as the substrate for GABA formation. The exceptional coupling of glutamine deamidation to glutamate decarboxylation in L. reuteri likely reflects adaptation to cereal substrates.

     

Metabolic response of porcine colon explants to in vitro infection by <Emphasis Type="Italic">Brachyspira hyodysenteriae</Emphasis>: a leap into disease pathophysiology

Introduction

Swine dysentery caused by Brachyspira hyodysenteriae is a production limiting disease in pig farming. Currently antimicrobial therapy is the only treatment and control method available.

Objective

The aim of this study was to characterize the metabolic response of porcine colon explants to infection by B. hyodysenteriae.

Methods

Porcine colon explants exposed to B. hyodysenteriae were analyzed for histopathological, metabolic and pro-inflammatory gene expression changes.

Results

Significant epithelial necrosis, increased levels of l-citrulline and IL-1α were observed on explants infected with B. hyodysenteriae.

Conclusions

The spirochete induces necrosis in vitro likely through an inflammatory process mediated by IL-1α and NO.

     

Elevated CO<Subscript>2</Subscript> and virus infection impacts wheat and aphid metabolism

Introduction

The aphid Rhopalosiphum padi L. is a vector of Barley yellow dwarf virus (BYDV) in wheat and other economically important cereal crops. Increased atmospheric CO₂ has been shown to alter plant growth and metabolism, enhancing BYDV disease in wheat. However, the biochemical influences on aphid metabolism are not known.

Objectives

This work aims to determine whether altered host-plant quality, influenced by virus infection and elevated CO₂, impacts aphid weight and metabolism.

Methods

Untargeted ¹H NMR metabolomics coupled with multivariate statistics were employed to profile the metabolism of R. padi reared on virus-infected and non-infected (sham-inoculated) wheat grown under ambient CO₂ (aCO₂, 400 µmol mol^?1) and future, predicted elevated CO₂ (eCO₂, 650 µmol mol^?1) concentrations. Un-colonised wheat was also profiled to observe changes to host-plant quality (i.e., amino acids and sugars).

Results

The direct impacts of virus or eCO₂ were compared. Virus presence increased aphid weight under aCO₂ but decreased weight under eCO₂; whilst eCO₂ increased non-viruliferous (sham) aphid weight but decreased viruliferous aphid weight. Discriminatory metabolites due to eCO₂ were succinate and sucrose (in sham wheat), glucose, choline and betaine (in infected wheat), and threonine, lactate, alanine, GABA, glutamine, glutamate and asparagine (in aphids), irrespective of virus presence. Discriminatory metabolites due to virus presence were alanine, GABA, succinate and betaine (in wheat) and threonine and lactate (in aphids), irrespective of CO₂ treatment.

Conclusion

This study confirms that virus and eCO₂ alter host-plant quality, and these differences are reflected by aphid weight and metabolism.

     

<Emphasis Type="Italic">massPix</Emphasis>: an R package for annotation and interpretation of mass spectrometry imaging data for lipidomics

Introduction

Mass spectrometry imaging (MSI) experiments result in complex multi-dimensional datasets, which require specialist data analysis tools.

Objectives

We have developed massPix—an R package for analysing and interpreting data from MSI of lipids in tissue.

Methods

massPix produces single ion images, performs multivariate statistics and provides putative lipid annotations based on accurate mass matching against generated lipid libraries.

Results

Classification of tissue regions with high spectral similarly can be carried out by principal components analysis (PCA) or k-means clustering.

Conclusion

massPix is an open-source tool for the analysis and statistical interpretation of MSI data, and is particularly useful for lipidomics applications.

     

Seroprevalence of <Emphasis Type="Italic">Borrelia burgdorferi</Emphasis> sensu lato and <Emphasis Type="Italic">Anaplasma phagocytophilum</Emphasis> in Danish horses

Background

Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum are able to infect horses. However, the extend to which Danish horses are infected and seroconvert due to these two bacteria is unknown. The aim of the present study was to evaluate the seroprevalence of B. burgdorferi sensu lato and A. phagocytophilum in Danish horses.

Methods

A total of 390 blood samples collected from all major regions of Denmark and with a geographical distribution corresponding to the density of the Danish horse population were analyzed. All samples were examined for the presence of antibodies against B. burgdorferi sensu lato and A. phagocytophilum by the use of the SNAP^®4DX ^® ELISA test.

Results

Overall, 29.0% of the horses were seropositive for B. burgdorferi sensu lato whereas 22.3% were seropositive for A. phagocytophilum.

Conclusions

Antibodies against B burgdorferi sensu lato and A. phagocytophilum are commonly found among Danish horses thus showing that Danish horses are frequently infected by these organisms.