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Biomedical subjects

J H Erkens

Publications and source records attributed to J H Erkens.

13 recordsLinked to original sources
PubMed2001

The major bovine mastitis pathogens have different cell tropisms in cultures of bovine mammary gland cells.

We previously showed that Staphylococcus aureus cells adhered mainly to an elongated cell type, present in cultures of bovine mammary gland cells. Moreover, we showed that this adhesion was mediated by binding to fibronectin. The same in vitro model was used here, to study adhesion of other important mastitis pathogens. Like the S. aureus strains, the Streptococcus dysgalactiae strains adhered mainly to elongated cells, which seemed to be mediated by fibronectin binding. In contrast, Streptococcus uberis strains adhered mainly to cubic cells. Since the cubic cells did not express fibronectin and S. uberis cells bound fibronectin less efficiently, the adhesion of S. uberis cells was independent of fibronectin binding. Streptococcus agalactiae strains adhered poorly to both cell types. The specificity and efficiency of adhesion of Escherichia coli strains was strongly strain dependent. None of the S. agalactiae and E. coli strains tested was able to bind fibronectin efficiently. The results suggest that the different mastitis pathogens have different target cell specificities and use different mechanisms to adhere to cells of the bovine mammary gland.

Animals↗
PubMed2001

Determination of the circadian rhythm in plasma corticosterone and catecholamine concentrations in growing broiler breeders using intravenous cannulation.

This paper describes a fast and simple technique for cannulation of the vena cutanea ulnaris (wing vein) in ad libitum fed growing broiler breeders of 5 weeks of age. Twenty-four hours after cannulation, blood was sampled every 4 h during 24 h. The circadian rhythm in plasma corticosterone and catecholamine concentrations was determined for the first time in growing broiler breeders. During the light period, baseline plasma corticosterone concentrations were higher than during the dark period, and reached the circadian peak at 11:00 h (lights on from 07:00 to 15:00 h). Plasma noradrenaline concentrations were higher during the light period than during the dark period, but there was no pronounced circadian rhythm in plasma adrenaline concentrations. Plasma corticosterone and catecholamine concentrations fell within the same range as previously reported in laying hens.

Animals↗
PubMed1999

Effects of repeated jugular puncture on plasma cortisol concentrations in loose-housed dairy cows.

In three experiments, the effects of venipuncture on plasma cortisol concentrations were studied in loose-housed dairy cows. In Exp. 1, two blood samples were collected 18 min apart on three alternate days from 20 dairy cows for studying their adrenocortical response to a single venipuncture. To further evaluate the effect of cows anticipating venipuncture, in Exp. 2, 15 dairy cows were sequentially venipunctured once daily on 12 successive days in a randomized order in groups of five, starting 15 min apart. In Exp. 3, 10 primiparous cows were used on three alternate days to study habituation to serial sampling (i.e., collection of five blood samples by venipuncture, 15 min apart). In cows accustomed to handling, jugular puncture did not affect cortisol concentrations in plasma collected 18 min later. Average daily cortisol concentrations varied between 2.07 +/- .38 and 3.81 +/- .56 ng/mL in the first (t = 0) and between 1.43 +/- .15 and 2.61 +/- .72 ng/mL in the second (t = 18) blood samples. Likewise, when cows were sampled sequentially once a day, the order of sampling between and within groups did not influence (P > .05) plasma cortisol concentrations. In contrast, primiparous dairy cows that were less used to being handled showed an average increase in cortisol concentrations when five samples were collected by venipuncture 15 min apart. During successive sampling sessions, however, the cows did not decrease or increase plasma cortisol concentrations in response to repeated serial sampling at the group level (P > .05). Between individuals, the maximum effect of repeated venipuncture on cortisol concentrations (4.5 to 22.6 ng/mL), the time at which the effect reached its maximum (30 to 60 min), and the consistency of the response pattern over successive series varied largely. The results of this study show that in cows that were accustomed to handling and to being restrained, baseline cortisol concentrations can be measured in single blood samples that are collected by jugular puncture within 1 min after first approaching the cow. When successive blood samples need to be collected within 15 to 20 min, jugular puncture may induce an increase in cortisol concentration, which seems to depend on the handling experience of the animals and on individual differences.

Animals↗
PubMed1998

New GnRH-like peptide construct to optimize efficient immunocastration of male pigs by immunoneutralization of GnRH.

Castration of male pigs is routinely performed in order to prevent the occurrence of boar taint in pig carcasses. However, boar taint can also be eliminated by immunological castration using a synthetic peptide vaccine against GnRH. For pig farming, to make immunocastration a feasible alternative method to surgical castration, the composition of the vaccine has to be not only reliable and effective but also cost-efficient and safe. Previously the authors have developed an effective immunocastration vaccine by replacing the monomer GnRH by a much more immunogenic tandem peptide. However, this tandem-GnRH vaccine preparation needs Complete Freund's adjuvant and to be applied at a relatively high dose. Therefore, alternative antigens were designed to cope with this problem and tested with different adjuvants and dosages. An effective new antigen was designed based on a GnRH-tandem peptide, which was dimerized and modified in one amino acid position of the decapeptide to allow conjugation of this tandem-dimer to ovalbumin. In mild adjuvants and in low dosage, this antigen was very effective in reducing testis weight, serum LH and androstenone level in backfat. Thus, an improved immunocastration vaccine has been designed that is relatively cost-efficient and highly efficacious in two vaccinations at low dose.

Adjuvants, Immunologic↗
PubMed1998

A time-resolved fluoroimmunoassay for cortisol in unextracted bovine plasma or serum with optimized procedures to eliminate steroid binding protein interference and to minimize non-specific streptavidin-europium binding.

A time-resolved fluoroimmunoassay (TR-FIA) for human salivary cortisol was adapted for the measurement of cortisol in unextracted bovine blood plasma and serum. It has been demonstrated that the binding of cortisol binding plasma proteins (CBPP) to the cortisol-biotin primary probe cannot be eliminated by means of cortisol releasing agents. Complete inactivation of CBPP was achieved by heating water diluted samples for 30 min at 80 degrees C. The high non-specific binding (NSB) of the streptavidin-europium secondary probe, encountered during preliminary experiments, was shown to be caused by an interaction with bovine serum albumin (BSA) and could be reduced partly by the addition of heparin. It was also shown that the ability to bind streptavidin-europium nonspecifically is not a general property of proteins since bovine gamma-globulin and gelatin lack this behaviour. The advantage of a highly reduced NSB, resulting from the use of a BSA free assay buffer, is not limited to this particular assay but is also beneficial for other procedures based on specific measurement of streptavidin-europium fluorescence. The detection limit for a 20 microl sample was 0.5 ng/ml. The intra-assay coefficients of variation for control samples with cortisol concentrations of 71.1, 39.2 and 10.3 ng/ml were 8.2, 7.9 and 11.3% (n = 16). The corresponding inter-assay coefficients of variation were 7.3, 9.0 and 11.2% (n = 73). Correlation with a commercially available radioimmunoassay, preceded by diethylether extraction of the sample, was 0.97 (n = 88).

Animals↗
PubMed1997

Genetic variation in the porcine myogenin gene locus.

The myogenin (MYOG) gene fulfills a key function in muscle differentiation by controlling the onset of myoblast fusion and the establishment of myofibers. In meat-producing animals like pigs and cattle, myofiber numbers have been related to growth capacity. We have characterized the porcine MYOG gene to detect genetic variation at this locus and to relate it to growth characteristics. MYOG gene fragments were isolated by PCR on genomic DNA and by screening a genomic library with a mixture of the four human MyoD cDNA fragments. Both the exons and promoter region were very similar to the human and mouse genes. Southern blot analysis of 105 unrelated pigs revealed three polymorphic MspI sites, located in the promoter region, the second intron, and at the 3' side of the gene. PCR-RFLP tests detecting four MYOG alleles were developed. PCR analysis of a panel of pig-rodent somatic cell hybrids confirmed the genetic localization of MYOG on pig Chromosome (Chr) 9. The PCR-RFLP tests and microsatellite markers on Chr 9 offer the possibility to genotype large numbers of pigs for studies of genetic linkage to meat deposition and growth characteristics.

Animals↗
PubMed1992

Isolation of sequences from a random-sequence expression library that mimic viral epitopes.

We describe the use of random peptide sequences for the mapping of antigenic determinants. An oligonucleotide with a completely degenerate sequence of 17 or 23 nucleotides was inserted into a bacterial expression vector. This resulted in an expression library producing random hexa- or octapeptides attached to a beta-galactosidase hybrid protein. Mimotopes, or antigenic sequences that mimic an epitope, were selected by immunoscreening of colonies with monoclonal antibodies, which were specific for antigenic sites on the spike protein of the coronavirus transmissible gastroenteritis virus. We report one mimotope for antigenic site II, eight for site III and one for site IV. The site III and site IV mimotopes were closely similar to the corresponding linear epitopes, localized previously in the amino acid sequence of the S protein. An alignment of the site II mimotope and the sequence of the S protein around Trp97, which is substituted in escape mutants, suggests that this mimotope mimics a conformational epitope located around residues 97-103. Applications of mimotopes to epitope mapping, serodiagnosis and vaccine development are discussed.

Amino Acid Sequence↗
PubMed1992

Development of a high capacity radioimmunoassay procedure: ovine follicle stimulating hormone determination in blood plasma as a model.

A sensitive, specific and accurate homologous radioimmunoassay (RIA) for ovine follicle stimulating hormone (oFSH) has been developed, using a [125I]oFSH tracer and a polyclonal rabbit anti-OFSH-serum at a final dilution of 1:224,000. The separation of free and antibody-bound tracer is based on the double antibody solid phase system. The assay was found to be specific for oFSH; cross-reactivity with oLH, oPrl and oGH was lower than 0.2%. The detection limit was 7 pg per tube. Inter- and intra- assay coefficients of variation (CV) were 3.7-8.5% and 6.6%, respectively. The use of a few selfmade appliances in combination with a well-considered time-schedule enabled the processing of three thousand blood plasma samples in triplicate within two weeks. The particular advantage of the method described here is the fast, easy and safe separation of free and antibody-bound tracer with minimal handling of radioactive tubes.

Animals↗
PubMed1992

Pubertal development in the male pig: effects of treatment with a long-acting gonadotropin-releasing hormone agonist on plasma luteinizing hormone, follicle stimulating hormone and testosterone.

The effects of a long-acting gonadotropin-releasing hormone (GnRH) agonist, [D-Trp6]-GnRH (GnRH-A) on developmental profiles of plasma luteinizing hormone (LH), follicle stimulation hormone (FSH) and testosterone (T), and pituitary responsiveness to exogenous GnRH were studied in male Dutch Landrace x Large White crossbred pigs from 1 to 30 wk of age. Group 1 control animals (control; n = 12) were injected subcutaneously in the neck with vehicle at 1 and 16 wk of age. Group 2 animals (early treatment; n = 10) were injected with 600 micrograms [D-Trp6]-GnRH at 1 wk and with vehicle at 16 wk. Group 3 animals (late treatment; n = 8) were injected with vehicle and 3 mg GnRH-A at 1 and 16 wk, respectively. Group 4 animals (early plus late treatment; n = 9) were injected at both 1 and 16 wk with GnRH-A. Blood was collected by brachiocephalic puncture at weekly or biweekly intervals, and through brachiocephalic cannulae, to determine longitudinal profiles of LH, FSH and T, and plasma gonadotropin responses to intravenous injection of GnRH (0.1 microgram/kg), respectively. In control animals, LH and FSH declined over the first 5 wk of postnatal life and peaked again at 10-14 wk. Levels of both hormones were basal from 18 to 30 wk. Plasma T was high in the first week, declined progressively over the next few weeks and remained low until 24 wk when a transient increment was noted. The LH and FSH responses to acute GnRH stimulation were similar at 7 and 14 wk and declined significantly at 23 wk of age.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Prolactin in the developing pig.

Prolactin (PRL) was determined in plasma of fetal pigs from 40 days post coitum (d.p.c.) onwards. Values increased in the last 3 wk of the gestational period and reached 10 ng/ml at term. Immunoreactive lactotropes could be identified as early as 31 d.p.c. Lactotrope development appeared to be biphasic, with no apparent increase in cell number in the 51-75 d.p.c. period. Sex differences in volume percentage of lactotropes and plasma PRL levels were observed in the 71-75 d.p.c. period. Pituitary PRL content increased 20-fold between 95 d.p.c. and term and 10-fold between birth and 6 wk. Volume percentage of lactotropes did not change from birth to 6 wk. In the postnatal period, plasma PRL concentrations were high in the first 1-2 wk and they decreased thereafter. Lower stable values were observed until about 10 wk, when PRL concentrations began to increase. Prepubertal peak levels were observed in the 10- to 16-wk period and a pubertal peak was observed at 20-22 wk. No significant differences between plasma PRL profiles of males and females were observed in the postnatal period. Castrated males, however, had consistently lower plasma PRL concentrations than intact boars. Cryptorchid boars had PRL concentrations comparable to intact boars. The PRL profile, in relation to the pattern of gonadal development, suggests a role for PRL in gonadal development of the pig.

Animals↗
PubMed1988

Effects of morphine and naloxone on plasma levels of LH, FSH, prolactin and growth hormone in the immature male pig.

The effects of acute i.v. administration of gonadotrophin-releasing hormone (GnRH; 0.1 micrograms/kg), morphine (3 mg/kg) and/or naloxone (0.5 mg/kg) on LH and FSH secretion was evaluated in young male pigs (approximately 6 weeks old) with venous brachiocephalic cannulae. The effects of morphine and/or naloxone treatments on prolactin and GH were also evaluated. The influence of morphine on hypophysial hormone secretion was also examined 2 days after castration. Animals treated with morphine and/or naloxone were compared with saline-injected control animals. Injection of GnRH induced 400 and 50% increases in LH and FSH respectively. Morphine and/or naloxone did not influence LH secretion in intact or castrated animals. Morphine suppressed (P less than 0.01) FSH levels 40-60 min after injection whereas naloxone had no effect. Castration eliminated morphine-induced suppression of FSH. Injection of morphine followed by naloxone resulted in acutely raised (P less than 0.05) FSH concentrations. Morphine induced a threefold increase (P less than 0.01) in prolactin within 30 min of injection and naloxone inhibited the effect of morphine. Levels of GH were increased (P less than 0.01) 20 min after morphine treatment and this increase was delayed when naloxone was given immediately after morphine. Naloxone alone did not affect prolactin or GH secretion. Castration caused increases in LH (P less than 0.05) and FSH (P less than 0.01), did not influence prolactin or GH, and reduced plasma testosterone to undetectable (less than 1.0 nmol/l) levels. These results suggest that in young male pigs the hypothalamic-hypophysial axis is responsive to GnRH and gonadal negative feedback.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Chronic GnRH administration in prepubertal male pigs. A model to evaluate the effects of GnRH treatment in cryptorchidism.

The effect of chronic pulsatile low-dose GnRH treatment on the juvenile testis and associated structures was evaluated in relation to hormonal parameters in the peripheral blood in the pig. Starting at 8 weeks of age, male pigs (crossbreds of Dutch Landrace and Yorkshire breeds) were injected 6 times daily im with 0, 75 or 250 ng GnRH/kg body weight during 4 weeks. Immediately after the treatment period, a GnRH stimulation test with 750 ng GnRH/kg iv was carried out. Samples for plasma LH, FSH, testosterone and 5 alpha DHT measurement were obtained weekly (basal level) and after GnRH stimulation. The pigs were castrated at 12 weeks of age and the weights and lengths of the testis, epididymis and cremaster muscle were recorded. Intratesticular testosterone and 5 alpha DHT concentrations were determined, and the testis and epididymis were evaluated for histological changes. Basal plasma hormone concentrations, intratesticular androgen concentrations and the response of the pituitary gland to stimulation had not been affected by GnRH treatment. Pigs receiving the higher treatment dose of GnRH showed less increase in testosterone levels in response to the stimulation dose at 12 weeks of age than the other pigs. Morphologically, no changes were observed in the epididymis and cremaster muscle after GnRH treatment and no signs of reactivation of structures that can provoke testicular descent could be seen. The development of the seminiferous epithelium was more advanced in the GnRH-treated groups, apparently in a dose-dependent manner.

Animals↗