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D Maes

Publications and source records attributed to D Maes.

At least 19 recordsLinked to original sources

Tetracycline-resistance in lactose-positive enteric coliforms originating from Belgian fattening pigs: degree of resistance, multiple resistance and risk factors.

Between March and October 2003 a field study was conducted in 50 randomly selected pig herds to assess the degree of tetracycline-resistance in lactose-positive enteric coliforms (LPEC) originating from fattening pigs and to evaluate the combined effects of various husbandry conditions on the development and persistence of antimicrobial-resistant bacteria. Data on housing, management and antimicrobial-drug consumption were collected, as well as faecal samples at three production stages: end of the nursery period (mean age: 72 days), end of the grower period (mean age: 125 days) and end of the finisher period (mean age: 186 days). The degree of tetracycline-resistant LPEC was determined by means of an agar dilution method. Tetracycline-resistant LPEC were found in every herd. The overall degree of tetracycline-resistance in LPEC was 56.8% (S.D. 22.4%). Only a very weak relation was found between the degrees of TETR in the different production stages within the same herd, indicating that the degree of TETR is mainly associated with the production stage rather than with the farm as a whole. The risk factor analysis showed that besides the antimicrobial-drug use, other factors like inside pen hygiene can influence the development and maintenance of antimicrobial-resistant bacteria in the gastrointestinal tracts of pigs. It was also observed that tetracycline-resistance in commensal Escherichia coli is often linked with resistance to other antimicrobial drugs like ampicillin and trimethoprim-sulphonamides. These results illustrate that the epidemiology of antimicrobial resistance is influenced by antimicrobial-drug use, cross-resistance development and non-antimicrobial risk factors.

Animal Husbandry↗

Protein variability among Mycoplasma hyopneumoniae isolates.

Sodium-dodecyl-sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was used to study the protein variability of Mycoplasma hyopneumoniae isolates. Fifty-six M. hyopneumoniae isolates from 6 different countries and 37 different herds were used. From eight herds, more than one isolate was available. All SDS-PAGE patterns of isolates originating from different herds were clearly divergent. Intra-species protein variability was quantified using the reference strain J and seven field strains all obtained from different herds and classified according to virulence. Between the field strains, a variability of 25% was found, while the culture-adapted strain J was clearly divergent and showed 30% variability with the field strains. No clustering according to virulence was obtained, but a protein band of about 181kDa was present in the two highly virulent isolates whereas this protein band was absent in the moderately and low virulent isolates. Protein patterns of isolates derived from different animals from the same herd, were identical or differed in only a few protein bands. This study clearly indicates that, in agreement with previous studies on genomic diversity of M. hyopneumoniae isolates, proteomic variability within the species is high. Our study did not find clear evidence that more than one M. hyopneumoniae isolate circulates within a herd at a specific time point. The minor differences found between M. hyopneumoniae isolates from the same herd might reflect the organism's ability to alter its proteomic expression profile under field conditions.

Animals↗

Interactions of highly and low virulent Mycoplasma hyopneumoniae isolates with the respiratory tract of pigs.

Mycoplasma hyopneumoniae is the etiological agent of swine enzootic pneumonia, a chronic nonfatal disease affecting pigs of all ages. To obtain better insight in the mechanisms responsible for differences in virulence between highly and low virulent M. hyopneumoniae isolates, 23 caesarean-derived, colostrum-deprived piglets were randomly assigned to three groups. Groups 1 and 2 consisted of nine animals each, which were intratracheally inoculated at 1 week of age with a highly or a low virulent isolate of M. hyopneumoniae, respectively. The remaining five animals were inoculated with sterile culture medium. Animals were euthanized at 5, 10, 15 and 28 days post-inoculation (DPI). Animals inoculated with the highly virulent isolate had more neutrophils in BAL fluid at 10, 15 and 28DPI compared to the other groups. At 10 and 15DPI, animals in the highly virulent group had significantly higher concentrations of TNF-alpha in BAL fluid. IL-1beta concentration in this group was higher at 5 and 28DPI compared to the other groups. From 10DPI onwards, significantly higher titres of M. hyopneumoniae were detected in the BAL fluid of animals inoculated with the highly virulent isolate compared to animals inoculated with the low virulent isolate. Additionally, the in vitro generation time of the highly virulent M. hyopneumoniae isolate was significantly shorter than that of the low virulent isolate. The present study indicates that the difference in pathogenicity between the highly and low virulent isolates is associated with a faster in vitro growth, a higher capacity to multiply in the lungs and the induction of a more severe inflammation process by the highly virulent isolate.

Animals↗

Developmental competence of bovine oocytes is not related to apoptosis incidence in oocytes, cumulus cells and blastocysts.

The number of follicles undergoing atresia in an ovary is very high, and isolation of cumulus-oocyte complexes (COCs) from such atretic follicles may impair subsequent embryo development in vitro. Our aim was to study if stringent selection by morphological assessment of COCs can improve embryo development, and to evaluate whether oocyte diameter is related with apoptotic ratio in oocytes and blastocysts. COCs from slaughtered cattle were recovered by follicle aspiration and classified depending on oocyte diameter: (A) <110 microm; (B) 110-120 microm; (C) >120 microm. COCs were matured, fertilized and cultured in vitro. Early and late stages of apoptosis were detected by Annexin-V and TUNEL staining, respectively, in denuded oocytes, COCs and blastocysts. Immature oocytes from Group A showed higher apoptotic ratio assessed by TUNEL assay, and the COCs corresponding to this group also showed a higher proportion of apoptotic cumulus cells. After maturation, no differences were present in the incidence of apoptosis among oocytes from different groups, but COCs corresponding to the largest diameter showed less apoptotic cumulus cells. In addition, the percentage of apoptotic oocytes decreased during in vitro maturation in all groups. Apoptotic cell ratio (ACR) in blastocysts was not related to oocyte diameter. In conclusion, oocyte selection and oocyte morphological evaluation prior to maturation was not sufficient to select non-atretic oocytes. When oocyte diameter was used as an additional selection the embryonic developmental potential increased together with oocyte diameter, but this improvement was not related to a lower incidence of apoptosis in the largest oocytes.

Animals↗

Comparison of sperm quality of Belgian Blue and Holstein Friesian bulls.

Few data are currently available on sperm quality of Belgian Blue (BB) bulls. The present study compared sperm quality of BB to Holstein Friesian (HF) bulls of several age categories, by means of a classical semen evaluation. Volume and concentration, and consequently total sperm output depended largely on age. Gross, total, and progressive motility, % live and % normal spermatozoa were significantly lower in the BB breed. Primary sperm abnormalities, such as nuclear vacuoles, midpiece defects and cytoplasmic droplets which were noticed most frequently, occurred far more in the BB breed. Hence, disturbances in spermiogenesis are deemed to be the cause of the poorer BB sperm quality. Since these sperm abnormalities occur significantly more in the BB breed than in the HF breed, it seems as if the BB breed is genetically predisposed to a higher susceptibility to environmental stresses which are known to interfere with normal spermiogenesis. The small scrota typical of the inbred BB breed might in part be responsible for this, and therefore selection for larger scrota in the BB breed is advisable.

Age Factors↗

A multiplex PCR to identify porcine mycoplasmas present in broth cultures.

Mycoplasma hyopneumoniae, Mycoplasma hyorhinis and Mycoplasma flocculare can be present in the lungs of pigs at the same time. These three mycoplasma species all require similar growth conditions and can be recovered from clinical samples using the same media. We have developed a multiplex PCR as a helpful tool for rapid differentiation of these three species in the course of isolation. Based on the 16S ribosomal DNA sequences, three different forward primers and a single reverse primer were selected. Each forward primer was compared to available mycoplasma sequences, showing the primers to be specific. The three amplification products observed of 1129 bp (M. hyorhinis), 1000 bp (M. hyopneumoniae) and 754 bp (M. flocculare) were clearly distinguishable on a 1% agarose gel. In addition, no cross-reaction with Mycoplasma hyosynoviae, another porcine mycoplasma, was noted. This multiplex PCR using the proposed set of primers is the first reported assay that allows the simultaneous identification of the different Mycoplasma species isolated from the lungs of pigs.

Animals↗

Variability in antimicrobial resistance among Salmonella enterica strains from fattening pigs and sows.

The aim of this study was to determine the prevalence of antibiotic resistance and different resistance patterns for Salmonella isolates collected from Belgian sows and fattening pigs at different ages and at slaughter. The most frequently isolated serotypes were S. Typhimurium (42.3%), S. Derby (25.1%), S. Goldcoast (7.3%), and S. Infantis (4.8%). All 901 isolates were submitted to antimicrobial susceptibility testing for 14 compounds using the disc agar diffusion test. The highest percentage of resistance was found to oxytetracycline (34.2%), streptomycin (32.5%), sulfamethizole (27.6%), and ampicillin (24.9%). Three of the isolates showed resistance to cephalosporins and none to second-generation fluoroquinolones. Multiresistance (resistance to > or =2 antimicrobials) was observed in 33.2% of the strains. Differences in resistance patterns were observed between and within serotypes as well as genotypes. A significant lower proportion (p < 0.01) of resistant strains was recovered in fecal samples from sows (23/56) than from fattening pigs during the weaning (30/30), growing (79/85), and finishing periods (45/52). The proportion of resistant strains recovered from fecal samples taken at the herd (154/167) was higher than in the samples collected at the slaughterhouse (75/140). When designing antimicrobial resistance surveillance programs for Salmonella in pigs, it is important to take multiple samples within each herd from both sows and fattening pigs at different time points.

Animals↗

Accuracy of trans-abdominal ultrasound pregnancy diagnosis in sows using a linear or sector probe.

The assessment of an early and reliable pregnancy diagnosis in sows is very important for limiting the number of non-productive days in sow herds. The aim of the present study was to investigate the accuracy of trans-abdominal ultrasound detection for early pregnancy diagnosis in sows under field conditions. Particular attention was paid to the time when a reliable diagnosis can be made, and to possible differences between two commonly used types of scanners. In total, 202 sows from two herds were tested daily with scanners A (linear transducer of 5 MHz) and B (sector transducer of 3.5 MHz) from day 16 until 25 after insemination. Ninety-three percentage of the sows were pregnant 36 days after insemination. Using ultrasound, the first pregnancy could be detected already 18 days after insemination. The sensitivity, specificity and overall accuracy were higher than 95% from day 24 (scanner A) or day 23 (scanner B) post-insemination onwards. The positive predictive values from day 19 onwards were consistently higher than 95% with both types of scanners, whereas the negative predictive values were consistently low for both scanners during the entire period. The sensitivity of testing tended to be slightly lower in sows with a litter size of <10 total born piglets, compared to sows with 10 or more piglets. In conclusion, pregnancy diagnosis in sows using trans-abdominal ultrasound testing can be assessed quickly and reliably under field conditions from day 23 of gestation onwards. Predictive values of positive test results were high in these herds, whereas those of negative test results were low. This implies that sows with a negative test result early in pregnancy should be retested later.

Abdomen↗

Prevalence of common gastrointestinal nematode parasites in scavenging pigs of different ages and sexes in eastern centre province, Burkina Faso.

The range and infestation intensities of gastrointestinal parasitic nematode species depend on the type of swine production system. The present study focused mainly on nematodes of veterinary importance in scavenging pigs in Burkina Faso, and aimed at determining the prevalence of gastro-intestinal nematode parasites by means of faecal egg per gram (EPG) counts. Between November 2001 and October 2002, faecal samples from 383 pigs of different sexes and ages (< 5 months, 5-12 months and > 12 months) were collected from the rectum and examined for gastrointestinal nematodes parasites using the Mc Master method. Of the 383 pigs examined, 91% were infected by one or more parasites. Ascaris suum (40%; 100-1 400 EPG) was the most prevalent parasite followed by Strongyloides ransomi (21%; 100-4200 EPG), Oesophagostomum spp. (18%; 100-1000 EPG), Hyostrongylus rubidus (11%; 100-1 800 EPG), Globocephalus spp. (10%; 100-400 EPG) and Trichuris suis (1 %; 100-200 EPG). The prevalence was significantly higher in female pigs (n = 239) than in males. In addition, females excreted significantly (P < 0.05) more eggs in their faeces than males, except in the case of Globocephalus spp. The age of the animal had no effect on the prevalence of A. suum whereas there were significant differences in age categories concerning S. ransomi, H. rubidus, Oesophagostumum spp. and Globocephalus spp. Unexpectedly, the high prevalence of these common parasites was not accompanied by elevated EPG values, which suggests the existence of moderate infestations. The present work indicates that the common nematode infestations in pigs do not necessarily need a systematic herd anthelmintic treatment, as only a small number of worms is required to induce immunity. A further study is needed to formulate appropriate and cost-effective strategies for the control of gastro-intestinal nematode parasites in pigs in Burkina Faso.

Age Factors↗

Breeding soundness and libido examination of Belgian Blue and Holstein Friesian artificial insemination bulls in Belgium and The Netherlands.

Data on breeding soundness and libido evaluations in Belgian Blue (BB) bulls are scarce. The present study compared results of breeding soundness and libido evaluations of young BB bulls to young Holstein Friesian (HF) bulls prior to acceptance into an AI program. Breed differences for breeding soundness exist between BB and HF bulls, as 93.7% of the young BB bulls failed the breeding soundness examination (BSE) compared to 59.3% of the HF bulls (P=0.0005). Within the BB breed, differences were present between bulls of different ages, and bull selection for better fertility with increasing age apparently influenced the results. The number of reasons for which bulls failed the test differed between the age groups in the BB breed, whereas a tendency for more failure reasons in the BB breed was noticed in the breed comparison. The most important reasons for failure were sperm morphology and scrotal circumference (SC), but far more BB bulls failed for these traits compared to the HF breed (82.8% versus 56.0% and 43.8% versus 17.6% in the BB and the HF breed for sperm morphology (P=0.0005) and SC (P<0.0001), respectively). The high proportion of BB bulls with a substandard SC and poor sperm morphology might suggest an increased prevalence of testicular hypoplasia or degeneration within this breed. Concerning libido, the reaction time did not differ either between breeds or between age groups within the BB breed, whereas mounting enthusiasm, although not different between the two breeds, did decline with increasing age, probably due to the greater mating experience of the older bulls. All in all, libido did not seem to be different between the breeds.

Age Factors↗

Effect of non-esterified fatty acids on bovine theca cell steroidogenesis and proliferation in vitro.

Elevated serum non-esterified fatty acid (NEFA) levels associated with a negative energy balance (NEB) may affect ovarian function and hence reproductive performance in high-yielding dairy cows. We have investigated the individual and combined effects of the three major NEFAs on bovine theca cell proliferation and steroidogenesis in vitro. Theca cells from healthy large follicles (>8 mm) obtained from slaughterhouse ovaries were cultured in serum free medium in the presence of 0, 50, 150 and 200 microM of palmitic acid (PA; C16:0); 0, 50, 150 and 250 microM of stearic acid (SA; C18:0); and/or 0, 50, 150 and 250 microM of oleic acid (OA; C18:1). Progesterone and androstenedione concentrations were measured in spent medium after 48 h of culture and cell numbers were determined spectrophotometrically per culture well. Cell viability was assessed by annexin-V FITC/propidium iodide staining. Only the treatment with 200 microM of PA inhibited cell proliferation (P<0.001) when tested individually, both of the mixtures tested (M1=100 microM of PA, 130 microM of SA and 140 microM of OA; M2=200 microM PA, 260 microM of SA and 280 microM of OA) reduced cell numbers (P<0.001). Progesterone and androstenedione production, both per well and per 10(4) cells, were not affected by any of the treatments, with the exception of M2. This mixture reduced progesterone production per well and per 10(4) cells (P<0.05). The effects observed were most likely caused by the cytotoxic action of the NEFAs, as demonstrated by the increased percentage of early apoptotic (M1) and late apoptotic/necrotic cells (M1 and M2) in the combination treatments (P<0.05). When combined, elevated physiological concentrations of PA, SA and OA can modulate theca cell proliferation and steroidogenesis in vitro by reducing theca cell viability. These NEFAs may be one of the mediators through which NEB compromises ovarian functioning and thus fertility in high-yielding dairy cows.

Androstenedione↗

Efficacy of in-feed medication with tylosin for the treatment and control of Mycoplasma hyopneumoniae infections.

The efficacy of in-feed medication with tylosin for the treatment of enzootic pneumonia was examined in an experimental Mycoplasma hyopneumoniae infection model. One group of 10 conventional M. hyopneumoniae-free pigs was inoculated intratracheally with a highly virulent field isolate of M. hyopneumoniae; a second group of 10 pigs was inoculated in the same way and after 12 days was given tylosin at 100 mg/kg feed for 21 days; a third group of 10 pigs was inoculated with sterile culture medium, and these pigs were not given tylosin. The pigs were examined daily for clinical signs and each pig was given a respiratory disease score. Thirty-three days after they had been infected the pigs were euthanased, the lung lesions were quantified and samples of lung were processed for immunofluorescence testing for M. hyopneumoniae. The mean (sd) respiratory disease and lung lesion scores were significantly higher (P<0.05) in both the infected groups than in the uninfected group. Between 23 and 33 days after infection the mean respiratory disease score of the pigs treated with tylosin was 0.54 (0.22), significantly (P<0.05) lower than that of the infected pigs which were left untreated, 1.54 (0.46); similarly, their average lung lesion score, 1.72 (1.20), was significantly lower than that of the untreated pigs, 5.27 (3.85).

Animal Feed↗

New techniques for the assessment of canine semen quality: a review.

Until recently, canine semen assessment was routinely performed by conventional light microscopic techniques. The limitations of these methods include subjectivity, variability, the small number of spermatozoa analyzed, and poor correlation with fertilizing potential. The last decade, several new in vitro techniques have been introduced for canine semen assessment that enable a more detailed evaluation of several sperm characteristics. Numerous fluorescent staining techniques have been developed for the evaluation of specific sperm characteristics and functions, including plasma membrane integrity, capacitation status and the acrosome reaction. By combining fluorescent stains, several functional sperm characteristics can be assessed simultaneously. Moreover, by means of flow cytometry, large numbers of fluorescently labelled spermatozoa can be analysed in a short interval. Following thorough standardization and validation, computer-assisted sperm analysis systems provide objective and detailed information on various motility characteristics and morphometric dimensions that cannot be identified by conventional light microscopic semen analysis. In vitro assays, evaluating the capacity of canine spermatozoa to bind to the zona pellucida or oviductal explants, or to penetrate the oocyte, provide additional information on canine gamete interaction that may be useful in predicting the fertilizing potential of spermatozoa. Although substantial improvements have been made in canine semen assessment, surprisingly few parameters were correlated with in vivo fertility. Therefore, further research is required to determine which sperm characteristics are of clinical value for predicting the in vivo fertility in dogs.

Acrosome Reaction↗

MYRRHA, a Pb-Bi experimental ADS: specific approach to radiation protection aspects.

Since 1998, SCK*CEN, in partnership with IBA s.a. and many European research laboratories, is designing a multipurpose accelerator driven system (ADS) for Research and Development (R&D) applications-MYRRHA-and is conducting an associated R&D support programme. MYRRHA is an ADS under development at Mol in Belgium and is aiming to serve as a basis for the European experimental ADS to provide protons and neutrons for various R&D applications. It consists of a proton accelerator delivering a 350 MeV x 5 mA proton beam to a liquid Pb-Bi spallation target that in turn couples to a Pb-Bi cooled, subcritical fast core. In the first stage, the project focuses mainly on demonstration of the ADS concept, safety research on sub-critical systems and nuclear waste transmutation studies. In a later stage, the device will also be dedicated to research on structural materials, nuclear fuel, liquid metal technology and associated aspects, and on sub-critical reactor physics. Subsequently, it will be used for research on applications such as radioisotope production. A first preliminary conceptual design file of MYRRHA was completed by the end of 2001 and has been reviewed by an International Technical Guidance Committee, which concluded that there are no show stoppers in the project and even though some topics such as the safety studies and the fuel qualification need to be addressed more deeply before concluding it. In this paper, we are reporting on the state-of-the art of the MYRRHA project at the beginning of 2004 and in particular on the radiation shielding assessment and the radiation protection particular aspects through a remote handling operation approach in order to minimise the personnel exposure to radiation.

Bismuth↗

Validation and usefulness of the Sperm Quality Analyzer (SQA II-C) for bull semen analysis.

In this study, an upgrade version of the Sperm Quality Analyzer (SQA), the SQA-IIC was tested for the assessment of bull semen quality. In Expt 1, the device showed good repeatability of measurements within and between capillaries, as evidenced by the low coefficients of variation (CVs; < 13%) at concentrations between 35 and 705 x 10(6) spermatozoa/ml. In Expt 2, 10 semen concentrations (1-1000 x 10(6)/ml) were stored in HEPES TALP for 48 h at room temperature. A time-dependent decrease in sperm motility index (SMI) values was noticed. SMI values increased linearly with increasing sperm concentrations, but remained constant around 500, corresponding to a concentration of approximately 50 x 10(6)/ml. For sperm concentrations below 50 x 10(6)/ml, SMI values were highly correlated with concentration (p < 0.05) and with semen parameters, expressing the overall semen quality (p < 0.05; Expt 3). In Expt 4, a correlation of only 0.44 (p < 0.05) between SMI values of frozen-thawed semen samples of 35 bulls and the corrected 56-day non-return rate (56dNRRc) was found. Prediction of the 56dNRRc based on the SMI value of a semen sample was inaccurate. The present study indicates that the SQA-IIC is suitable for a rapid screening of bull semen diluted to a concentration of approximately 50 x 10(6)/ml. Furthermore, the device seems inappropriate for fertility prediction.

Animals↗

Effect of non-esterified fatty acids on bovine granulosa cell steroidogenesis and proliferation in vitro.

In high-yielding dairy cows, the negative energy balance (NEB) during the first weeks post partum may influence dominant follicle growth and steroidogenesis. Since non-esterified fatty acid (NEFA) concentrations are elevated during NEB and are shown to be toxic for several cell types, we investigated the individual and combined effects of the three main NEFA's on granulosa cell proliferation and steroidogenesis in vitro. Granulosa cells from large follicles were cultured for two days in serum free medium in the presence of palmitic (C16:0) (PA), stearic (C18:0) (SA) and/or oleic acid (C18:1) (OA). Addition of 150, 300 or 500 microM of PA and SA inhibited cell proliferation (P<0.05) while OA only elicited such an effect at 500 microM (P<0.01). In the combination treatment (150 microM of each fatty acid), cell numbers were also reduced (P<0.01). These inhibitory effects on cell number are partly due to the induction of apoptosis by these NEFA's, as was demonstrated by annexin V-FITC/propidium iodide staining of the granulosa cells. Oestradiol-17beta production was stimulated by all doses of PA, by 300 and 500 microM of SA and by 500 microM of OA (P<0.05). Combined treatment with 150 microM of each fatty acid also stimulated oestradiol-17beta production per 10(4) cells (P<0.05). We can conclude that PA, SA and to a lesser degree OA modulate granulosa cell proliferation and steroidogenesis in vitro. These effects may be involved in the occurrence of ovarian dysfunction during the postpartum period in high-yielding dairy cows.

Animals↗

Risk factors for the herd-level bacteriologic prevalence of Salmonella in Belgian slaughter pigs.

Herd-level risk factors for salmonellosis in pigs were investigated in a cross-sectional study on 62 Belgian farrow-to-finish pig herds belonging to one slaughterhouse cooperative. Data concerning housing and ventilation, management, hygiene, biosecurity, production parameters, feeding, disease control and transport to the slaughterhouse were collected during a herd visit by means of a questionnaire. The percentage of positive animals in a slaughterhouse delivery, as determined by qualitative Salmonella isolation in the mesenteric lymph nodes taken from 30 slaughter pigs, was the outcome variable. All samples were taken in 4 different slaughterhouses. Variables first were submitted to a univariable analysis using a logistic mixed regression model, with herd as random effect. Variables which were related to the Salmonella prevalence (P < 0.05) were analysed further in a multivariable model. The clustering of Salmonella infection within a pen also was studied in a generalised mixed model with pen as random effect. Salmonella isolates were identified by serotype. In 57 (92%) of the herds, at least one sample was found positive for Salmonella. The median percentage of positive Salmonella samples per delivery was 64% (range: 0-100%). In the multivariable model, only type of floor was related significantly to the prevalence: 100% (95% CI 88-100) for herds with <50% slatted floors to 54% (36-70) for herds with fully slatted floors. The results from the analysis should be interpreted with care because only 62 herds were included in the study. Clustering between pigs from the same pen could not be demonstrated (variance +/- S.D.: 0.11 +/- 0.16). S. typhimurium (30%) and S. derby (20%) were most common among the 23 different serotypes that were found.

Abattoirs↗

Structural study of the type II 3-dehydroquinate dehydratase from Actinobacillus pleuropneumoniae.

The structure of the type II dehydroquinate dehydratase (DHQase) from Actinobacillus pleuropneumoniae, the third enzyme of the shikimate pathway, has been determined. Crystals diffracting to 1.7 A were obtained in space and on earth using the counter-diffusion technique. The structure was solved using molecular replacement and refined to high resolution. The overall structure of the dodecameric enzyme is described and compared with structures of DHQases from other bacteria. DHQases contain a flexible loop that presumably closes over the active site upon substrate binding. The enzyme can exist in an open or closed conformation. The present structure displays the open conformation, with a sulfate anion bound in the active site. The availability of this structure opens a route to structure-based antibiotics targetting this pathogenic bacterium.

Actinobacillus pleuropneumoniae↗