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Biomedical subjects

A De Kruif

Publications and source records attributed to A De Kruif.

At least 19 recordsLinked to original sources

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↗

Evaluation of the epidemiological importance of classical swine fever infected, E2 sub-unit marker vaccinated animals with RT-nPCR positive blood samples.

It has been demonstrated that pigs that have been double vaccinated with an E2 sub-unit marker vaccine and that are infected with classical swine fever virus (CSFV) through a natural contact infection may react positive in a CSFV detecting RT-nPCR test, whereas no virus could be isolated by using the conventional virus isolation (VI) technique. To evaluate whether these vaccinated and infected pigs may spread the virus, three experiments were set up. In the first, susceptible pigs were inoculated with serum originating from vaccinated RT-nPCR positive pigs. In the second, vaccinated RT-nPCR positive pigs were brought into contact with sentinel animals. In the third, vertical transmission was evaluated in RT-nPCR positive vaccinated pregnant gilts. In the first two experiments, no proof of virus transmission was found, whereas in the third vertical transmission was observed. The conclusion is that in vaccinated pigs that are positive in RT-nPCR but negative in VI, the level of circulating virus is probably not high enough for horizontal transmission, whereas vertical transmission of the virus is possible.

Animals↗

Inhibition of bovine sperm-oocyte fusion by the p-aminophenyl derivative of D-mannose.

Several steps in the process of mammalian fertilization are mediated by carbohydrates. This study investigated the role of the p-aminophenyl derivative of d-mannose (APMP) during bovine fertilization. Inseminating cumulus-oocyte complexes (COCs) in the presence of increasing APMP concentrations resulted in a significant dose-dependent decrease of the fertilization rate (P < 0.05). No negative effect of 50 mM APMP on total sperm motility and progressive motility was found. Subsequently, the fertilization steps at which this blocking effect could be exerted were investigated, i.e., sperm penetration of the cumulus oophorus, sperm-zona binding, acrosome reaction, sperm-oolemma binding, and/or sperm-oocyte fusion. Inseminating cumulus-enclosed and cumulus-denuded oocytes in the presence of 50 mM APMP significantly decreased the fertilization rate to a comparable minimum level (P < 0.05). There was no significant relationship between the number of spermatozoa bound to the zona pellucida and the APMP concentration, and APMP nor d-mannosylated bovine serum albumin (BSA) suppressed or stimulated sperm acrosomal status. Inseminating zona-free oocytes in the presence of 50 mM APMP did not influence sperm-oolemma binding, but significantly inhibited sperm-oocyte fusion (P < 0.05). Preincubating zona-free oocytes with 200 microg/ml Con A but not with 50 mM APMP inhibited the sperm-oocyte fusion rate to the same extent as when the gametes were simultaneously exposed to 50 mM APMP. These data indicate that the blocking effect of APMP on bovine fertilization is mainly due to an inhibition of sperm-oocyte fusion, probably by specific obstruction of the sperm receptor sites that are responsible for the fusion process.

Acrosome Reaction↗

Antimicrobial resistance in livestock.

Antimicrobial resistance may become a major problem in veterinary medicine as a consequence of the intensive use and misuse of antimicrobial drugs. Related problems are now arising in human medicine, such as the appearance of multi-resistant food-borne pathogens. Product characteristics, dose, treatment interval and duration of treatment influence the selection pressure for antimicrobial drug resistance. There are theoretical, experimental and clinical indications that the emergence of de novo resistance in a pathogenic population can be prevented by minimizing the time that suboptimal drug levels are present in the infected tissue compartment. Until recently, attention has been focused on target pathogens. However, it should be kept in mind that when antimicrobial drugs are used in an individual, resistance selection mainly affects the normal body flora. In the long term, this is at least equally important as resistance selection in the target pathogens, as the horizontal transfer of resistance genes converts almost all pathogenic bacteria into potential recipients for antimicrobial resistance. Other factors contributing to the epidemiology of antimicrobial resistance are the localization and size of the microbial population, and the age, immunity and contact intensity of the host. In livestock, dynamic herd-related resistance patterns have been observed in different animal species.

Animals↗

Non-infectious factors associated with macroscopic and microscopic lung lesions in slaughter pigs from farrow-to-finish herds.

A cross-sectional epidemiological study was conducted in 150 randomly selected farrow-to-finish herds to investigate which non-infectious factors might act as risk indicators for the prevalence and severity of macroscopic and microscopic lung lesions in slaughter pigs. Data were collected during herd visits through inspections of the pigs and through interviews with the farmers. Macroscopic lung lesions of pneumonia and pleuritis were recorded at slaughter from 25 pigs per herd, and microscopic lung lesions of lymphohistiocytic infiltration were recorded from 10 pigs per herd. The median herd level prevalences were 24 per cent for pneumonia, 16 per cent for pleuritis and 60 per cent for lymphohistiocytic infiltration. Pneumonia lesions were negatively associated with pleuritis lesions and positively associated with lymphohistiocytic infiltration. Pleuritis lesions were negatively associated with lymphohistiocytic infiltration. The prevalence and the severity of pneumonia lesions were increased by a high frequency of purchasing gilts and by a slaughter date in January to February. The presence of a growing unit also increased the severity of pneumonia. The prevalence and the severity of pleuritis lesions were higher when there were more pig herds in the municipality, and when there were poor biosecurity measures, and their prevalence was increased by a slaughter date in January to February, and their severity by a slaughter date in March to April. An increase in the airspace stocking density in the finishing unit also increased the prevalence of pleuritis. The prevalence and the severity of lymphohistiocytic infiltration in the lung tissue were higher in herds purchasing gilts. Pigs raised in pens with slatted floors were also at higher risk of more severe lesions of lymphohistiocytic infiltration.

Animal Husbandry↗

Descriptive epidemiology of a classical swine fever outbreak in the Limburg Province of Belgium in 1997.

This paper describes the epidemiological characteristics of the 1997 Classical Swine Fever (CSF) outbreak that occurred in the Limburg Province of Belgium, where there is a policy of non-vaccination, intensive surveillance and eradication. Between 30 June and 17 July 1997, eight herds, located in three different areas, were confirmed to be CSF-positive. CSF virus was transmitted from the primary infected herd of one area to another five herds in the same area and to one herd in a different area. The mode of virus introduction for this primary infected herd and for the one herd that was not infected by this primary herd could not be determined. Clinical, serological, and virological findings indicated that the CSF-infected herds were detected in an early stage of the infection. The early detection of the infection together with a preventive stamping out procedure resulted in a rapid elimination of the CSF virus. A total of 46,561 pigs were slaughtered to control the spread of the infection. Another 27,579 pigs were slaughtered in the framework of the market support. The total direct costs of the episode were estimated at [symbol: see text] 10,893,337.

Animals↗

An experimental infection with classical swine fever virus in pregnant sows: transmission of the virus, course of the disease, antibody response and effect on gestation.

An experimental infection with classical swine fever (CSF) virus in 12 conventional gilts, housed in a sow-box housing system, was conducted in order to evaluate horizontal transmission, clinical, virological and serological response, and the effect on gestation. Two of the 12 gilts, of which 10 were pregnant, were experimentally inoculated. They became viraemic for the first time 6 days post-inoculation (dpi). The contact gilts became viraemic between 18 and 21 days post inoculation. On the basis of virological findings and the martingale estimate of R0 (13.0) it was concluded that the two experimentally inoculated gilts infected all contact gilts, although random contacts between gilts were not possible. The prescence of CSF infection could be diagnosed earlier and during a longer period when the leucocyte count or polymerase chain reaction were used in comparison with virus isolation in whole blood (P < 0.05). The observed clinical symptoms were atypical and highly variable between the gilts, which hampered clinical diagnosis. The pregnant gilts became infected between day 43 and 67 of gestation. In all cases vertical virus transmission occurred and this resulted partially in abortion and/or mummification.

Abortion, Veterinary↗

Chemiluminescence of bovine polymorphonuclear leucocytes during the periparturient period and relation with metabolic markers and bovine pregnancy-associated glycoprotein.

The respiratory burst activity of polymorphonuclear leucocytes (PMN) was evaluated in eight Holstein cows from 8 weeks before until 6 weeks after calving by chemiluminescence (CL). The CL response started to decrease 1 week before parturition, reaching a minimum during the first 2 weeks after calving. From week 3 of lactation, CL increased again and returned to original levels by week 6 of lactation. Plasma concentrations of 3-hydroxybutyric acid, total bilirubin and bovine pregnancy-associated glycoprotein started to increase before parturition to reach a maximum during the first or second week of lactation. The concentrations of glutamic-oxaloacetic transaminase, lactate dehydrogenase, non-esterified fatty acids and bilirubin increased after calving, reaching a maximum during the second week. A small decrease in plasma cholesterol during the week before and after calving was followed by an increase. The CL response of the PMN showed significant temporal relationships with the plasma concentrations of 3-hydroxybutyric acid, bovine pregnancy-associated glycoprotein, bilirubin, glutamic-oxaloacetic transaminase, non-esterified fatty acids; that with cholesterol was nearly significant. This means that the change in the CL response with time coincided with the changes in plasma concentrations of these substances with time and that these changes were significantly related with each other. The results of this study show that the decreased respiratory burst activity of bovine PMN around parturition may be related to the extent of the metabolic and hormonal changes. Although the causative relationships are not proven, these results support earlier results suggesting that 3-hydroxybutyric acid and bovine pregnancy-associated glycoprotein may directly affect neutrophil function, whereas non-esterified fatty acids, cholesterol, bilirubin, and liver enzymes may have potential as diagnostic markers of impaired neutrophil function and consequently increased disease risk around parturition.

3-Hydroxybutyric Acid↗

Effect of bovine herpesvirus-1 or bovine viral diarrhea virus on development of in vitro-produced bovine embryos.

In previous experiments, zona pellucida (ZP)-intact in vitro-produced (IVP) embryos incubated for 1 hr with 10(6.3) TCID(50)/ml bovine herpes virus-1 (BHV-1), 10(5.3) TCID(50)/ml cytopathic (CP) bovine viral diarrhea virus (BVDV) or 10(5.3) TCID(50)/ml noncytopathic (NCP) BVDV showed no signs of virus replication or embryonic degeneration. The aims of the present study were to investigate whether a prolonged presence (24 hr or 8 days) of 10(6.3) TCID(50)/ml BHV-1 or 10(5.3) TCID(50)/ml BVDV in an in vitro embryo production system affected the rate of cleavage and embryonic development of ZP-intact embryos, and to point out eventual causes of adverse effects. When virus was present in each step of an IVP system, significantly lower rates of cleavage and blastocyst formation of virus-exposed embryos were observed, in comparison with control embryos (P < 0.01). When embryos were only exposed to virus during the in vitro fertilization (IVF), the rates of cleavage and blastocyst formation were significantly affected. The introduction of BHV-1 or BVDV during in vitro maturation (IVM) or in vitro culture (IVC) resulted only in significantly lower rates of blastocyst (P < 0.01). In all experiments, virus replication was not detected in the embryonic cells. On the other hand, virus replication was clearly demonstrated in oviductal cells in the co-culture system, resulting in a degeneration of these cells. In an additional experiment, synthetic oviduct fluid (SOF) without somatic cells was used as an alternative culture system. Even when SOF-embryos were exposed to 10(6.3) TCID(50)/ml BHV-1 or 10(5.3) TCID(50)/ml CP, and NCP BVDV, the rates of blastocyst formation of the BHV-1-, CP-, and NCP BVDV-exposed embryos were not different from the unexposed control embryos, 23%, 24%, and 24%, respectively, vs. 27%. Taken together, it can be concluded that the virus-induced adverse effects on embryonic development in conventional co-cultures were due to changes in the embryonic environment caused by infection of oviductal cells.

Animals↗

Successful treatment of an equine preputial fibrosarcoma using 5-fluorouracil/evaluation of the treatment using quantitative PCNA and Ki67 (MIB 1) immunostaining. (case report).

An 18-month-old cross-bred Fjord pony colt presenting a fibrosarcoma of the prepuce with severe infiltrative growth, was treated by topical application of 5-fluorouracil. Biopsies were taken before and after a treatment period of 2 months. As a control, preputial tissue from a healthy horse was biopsied. Tissue sections were stained immunohistochemically for Proliferating Cell Nuclear Antigen (PCNA) and Proliferation related factor--Ki67 (MIB1). Using computerized quantitative image analysis of these sections, Mitotic index (PCNA), Growth Fraction (Ki67) and total nuclear area percentage of the dermis were calculated. The Mitotic index, growth fraction and total nuclear area percentage were significantly (P < 0.001) reduced at the end of the treatment period compared to the initial biopsy tissue, but they still were significantly higher than the control values, due to a remnant inflammatory reaction to the cytostatic drug. After treatment of the lesion, the pony fully recovered with no visible residual lesions on the prepuce. On follow-up 6 months after treatment, the horse showed no signs of recurrence. The success of the present treatment should encourage further clinical trials in cases of malignant fibrous tumours of skin in horses. The quantitative analysis of cell proliferation in biopsies can be used to evaluate treatment follow-up.

Animals↗

HPLC on Chiralcel OJ-R for enantiomer separation and analysis of ketoprofen, from horse plasma, as the 9-aminophenanthrene derivative.

Racemic ketoprofen is a non-steroidal anti-inflammatory drug used to treat musculoskeletal and colic conditions in horses. The enantioselective chiral inversion of ketoprofen administered to horses has been studied by use of cellulose tris(4-methylbenzoate), also known as Chiralcel OJ-R, as chiral stationary phase; acetonitrile - 0.02 M perchlorate buffer (pH 2.0)-methanol, 60:15:25 (v/v/v) was used as mobile phase. Before chromatography, to effect adequate chiral interaction with the chiral stationary phase ketoprofen was derivatized with 9-aminophenanthrene, under acid conditions, after solid-phase (C18) extraction and then liquid-liquid extraction, to ensure effective removal of endogenous plasma materials. The 9-aminophenanthrene derivative of S-ibuprofen was used as internal standard. The enantiomers of ketoprofen were separated to baseline (Rs = 6.44, alpha = 1.76) within a short analysis time. The results indicate that the bio-inversion of R-ketoprofen to the S isomer is significant in equine species. However, considerable differences in pharmacokinetic parameters were observed, indicating large inter-animal variation.

Animals↗

Influence of parity and stage of lactation on the somatic cell count in bacteriologically negative dairy cows.

This study examines the influence of parity, stage of lactation, and single isolations (i.e., the isolation of a microorganism that could not be reisolated in the same quarter in the next sampling) of staphylococci other than Staphylococcus aureus (coagulase-negative staphylococci), Corynebacterium bovis, or esculin-positive cocci other than Streptococcus uberis (referred to as esculin-positive cocci throughout) on the monthly log(e)-transformed somatic cell count (SCC) for 180 first, second, and third parity cows that were observed over a whole lactation. Repeated measures ANOVA was used to analyze the data. No significant effect was found for the infection variable. However, the results indicated that even single isolations of coagulase-negative staphylocci, C. bovis, or esculin-positive cocci resulted in a numerical or statistically significant increase in SCC. Least squares mean SCC (log(e)-transformed) for bacterio-logically negative cows and cows with single isolations of coagulase-negative staphylococci, C. bovis, or esculin-positive cocci were 3.90, 3.97, 4.08, and 4.17, respectively. Significant effects of parity, stage of lactation, and the interaction of parity and stage of lactation could not be found when only bacteriologically negative cows were considered. Least squares mean SCC for first, second, and third parity cows were 3.80, 3.93, and 3.97, respectively. However, the effects of parity, stage of lactation, and the interaction of parity and stage of lactation were significant when all 180 cows were included. Therefore, these effects must be due to factors that were present in the infected groups.

Animals↗

Cell allocation to the inner cell mass and the trophectoderm in bovine embryos cultured in two different media.

Data from other laboratories have shown that speed of bovine blastocyst development is higher when Ménézo B2 is used for coculture compared to TCM199. It was our purpose to investigate whether this early blastocyst formation was also indicative of embryo quality by studying the allocation of inner cells in embryos generated by B2-coculture and by TCM199-coculture. For this purpose, a differential staining technique was used. General embryo development was similar for TCM199- and B2-embryos expressed as rate of cleavage at day 3 and morula-blastocyst formation at day 8 (P > 0.05), but significantly different when expressed as number of eight-cell stages at day 3 and expanded or hatched blastocysts at day 8 (P < 0.01). B2-embryos cultured until day 5, 6, and 7 post insemination, had total cell numbers of 24, 65, and 109 respectively, which was significantly higher than the cell number of TCM199-embryos cultured over the same time period (18, 41, and 71 respectively, P < 0.001). Morphological differentiation was significantly more advanced for B2-embryos at day 7 and 8 (P < 0.0001 and P < 0.001, respectively). First presumptive inner cells appeared in eight- to 16-cell stages at day 3. Because the determination of inner cells by differential staining is depending upon the presence of functional tight junctions, we concluded that the establishment of the tight junction seal in B2-embryos differed from that in TCM199-embryos: Inner cells appeared 0.56 cell cycle later in B2-embryos (P < 0.001) and a larger variation existed in the number of ICM-cells in B2-blastocysts (P < 0.001). The higher total cell number of B2-expanded blastocysts was mainly acquired by trophectoderm growth (P < 0.06). These data indicate that the apparent better quality of B2-embryos (faster cleavage, earlier blastocyst formation) is not reflected in a reliable number of inner cells of B2-blastocysts.

Animals↗

[Perinatal mortality and the birth of weak calves].

In this review article the problem of perinatal mortality and the birth of weak calves is discussed. Two main causes can be distinguished: hypoxia and infections. It is emphasized that for a correct diagnosis a combination of a comprehensive anamnesis, a thorough examination of the calf and a laboratory investigation is necessary. Treatment of weak calves is in many cases disappointing. Preventive measures as an excellent management of periparturient cows are important.

Animals↗