Transmission of Salmonella between wildlife and meat-production animals in Denmark.

AIMS: To investigate the transmission of Salmonella spp. between production animals (pigs and cattle) and wildlife on production animal farms in Denmark. METHODS AND RESULTS: In the winter and summer of 2001 and 2002, 3622 samples were collected from Salmonella-infected and noninfected herds of pigs and cattle and surrounding wildlife. Salmonella was detected in wildlife on farms carrying Salmonella-positive production animals and only during the periods when Salmonella was detected in the production animals. The presence of Salmonella Typhimurium in wild birds significantly correlated to their migration pattern and food preference. CONCLUSIONS: Salmonella was transmitted from infected herds of production animals (cattle and pigs) to wildlife that lived amongst or in close proximity to them. SIGNIFICANCE AND IMPACT OF THE STUDY: Salmonella in animal food products is associated with the occurrence of Salmonella in primary animal production. Strategies to control the introduction and spread of infection should include wildlife management, as the nearby wildlife may act as reservoirs for Salmonella spp. and/or may be passive carriers of the bacteria.

Demonstration of neuropeptide Y and its precursor in plasma and follicular fluid.

The present investigation provides three lines of evidence for the presence of a pro-form of neuropeptide Y (NPY) in plasma and follicular fluid. First, by the demonstration of NPY-immunoreactive material of a size corresponding to the estimated mol wt of pro-NPY. Second, an antiserum specific for the C-terminal tyrosine amide of NPY and peptide YY does not react with this material. Third, it was possible to convert the pro-NPY extracted from plasma and follicular fluid using the protease, Endoproteinase-Lys C, to a NPY-immunoreactive form eluting slightly before NPY on a G-50 column. The size of the digested product was consistent with a cleavage of pro-NPY resulting in an immunoreactive species, NPY-Gly-Lys. Pro-NPY was also found in tissue culture media from the human neuroendocrine cell line SH-SY5Y. As in the case of plasma and follicular fluid, another NPY immunoreactive species eluted from a G-50 gel filtration column slightly before synthetic human NPY. Analysis of this material with an antibody directed against the tyrosine amide of NPY in combination with isoelectric focusing revealed that this peak consisted of at least two immunoreactive forms of NPY. In conclusion, at least three different forms of NPY immunoreactivity are likely to be present in plasma, follicular fluid, and cell tissue culture media; pro-NPY, a degradation form of pro-NPY, or a biosynthetic intermediate and NPY.

The effect of neuropeptides on vessel tone and cAMP production.

The effect of VIP, PHM, PHV, PACAP-27, and PACAP-38 on vessel tone and cAMP production was investigated in rabbit ovarian arteries in vitro. The peptides (10(-7)M) induced a significant relaxation on NA-precontracted vessels and displayed similar potencies. The cAMP accumulation induced by PACAP-27 and PACAP-38 was five times higher than the cAMP content induced by VIP, PHM, and PHV. NPY (10(-7)M) markedly reversed the relaxations induced by VIP, PHM, PHV, PACAP-27, and PACAP-38 but did not at all affect the cAMP production induced by these peptides. We conclude that the relaxation induced by VIP, PHM, PHV, PACAP-27, and PACAP-38 and the contraction evoked by NPY are not solely related to the changes of cAMP contents, and that in addition to cAMP, another intracellular signal transduction pathway may be involved.

Severity of illness and outcome of treatment in alcoholic patients in the intensive care unit.

To study whether critically ill alcoholics were more sick and had a worse outcome than other patients treated in the intensive care unit, data were collected during the initial 24 h on 216 consecutive patients admitted to an intensive care unit. Twentysix patients (12%) met the criteria for alcohol abuse. The patients' chronic health 6 months prior to admission and the extent of physiological derangement (Acute Physiology Score and Chronic Health Evaluation (APACHE] were recorded just as the type and amount of treatment (Therapeutic Intervention Scoring System (TISS]. Alcoholics were found to be significantly more sick and had a higher mortality (50% compared to 26%) than other critically ill patients. However, when analyzing the TISS points, no difference was found between the two groups. All survivors were, every third month up to 1 year after admission, asked to fill in a questionnaire indicating their level of activity. No differences were found between the two groups 1 year after admission, but the alcoholics had lost more time due to death. It is concluded that studies with larger number of patients will reveal whether alcoholics constitute a special category of patients with a different prognosis than other ICU patients.

Interaction between norepinephrine, NPY and VIP in the ovarian artery.

The in vitro effect and the interaction between norepinephrine (NE), neuropeptide Y (NPY) and vasoactive intestinal peptide (VIP) were studied in dissected segments of the rabbit ovarian artery. In addition, the structural requirement of the NPY receptor was investigated using NPY peptide analogs. NE induced a dose-dependent vasoconstriction with an Emax of 131.4 +/- 2.9% of K(+)-induced constriction. The vasoconstrictor effect of NPY was less than 5% of K(+)-induced vasoconstriction. Incubation of the artery with 10(-7) M NPY for 4 min induced a significant potentiation of NE-induced contractions. The selective NPY Y1 receptor agonist [Leu31, Pro34]NPY was also able to potentiate the NE response at the half-maximum contraction level, but not NPY(11-36), an NPY peptide fragment predominantly stimulating the NPY Y2 receptor. NPY exerted a dose-dependent vasoconstrictor effect on vessels contracted for 20 min with 10(-6) M NE. VIP induced a dose-dependent relaxation of vessels contracted with 10(-6) M NE. The VIP-induced relaxation could be reversed by NPY. In conclusion, receptors capable of interacting with NPY, presumably of the Y1 type, and VIP are present in the rabbit ovarian artery, and activation of these receptors may profoundly influence the response of the artery to norepinephrine.

Neuropeptide Y reduces ovarian blood flow in the rabbit.

Neuropeptide Y-containing nerve fibers have previously been demonstrated to innervate the mammalian ovary. These nerve fibers innervate primarily the vasculature. In this study we have developed a method for in vivo measurement of the ovary blood flow rate by means of the 133Xe method. Using this technique we measured the ovary blood flow rate and investigated the dose-response relationship between close intraarterial-injected NPY and the ovary blood flow rate. A monoexponential washout curve for 133Xe was found for the whole washout process, ensuring that the blood flow rate at any time could be calculated from the curve. We found a mean blood flow rate in the nonpregnant rabbit ovary at 43.6 +/- 4.4 ml.(100 g)-1.min-1 (mean +/- SEM). Injection of NPY (20, 200, 2000 pM) in the aorta close to a. ovarica resulted in a dose-dependent decrease in the ovarian blood flow rate with a maximum reduction to 40.7 +/- 6.3% (mean +/- SEM) of the control blood flow rate. These findings make it likely that receptors able to interact with NPY are present in the vasculature of the rabbit ovary.

Vascular effects and cyclic AMP production produced by VIP, PHM, PHV, PACAP-27, PACAP-38, and NPY on rabbit ovarian artery.

The relationship between vessel tone and cAMP production induced by vasoactive intestinal polypeptide (VIP), peptide histidine methionine (PHM), peptide histidine valine (PHV), pituitary adenylate cyclase activating polypeptide (PACAP-27 and PACAP-38), and neuropeptide Y (NPY) was investigated in rabbit ovarian arteries in vitro. VIP, PHM, PHV, PACAP-27, and PACAP-38 added in single-dose experiments (10(-9), 10(-8), 10(-7), and 10(-6) M) induced all a significant dose-related relaxation of noradrenaline (NA)-precontracted vessels and displayed similar potencies. VIP, PHM, PHV, PACAP-27, and PACAP-38 all increased cyclic adenosine monophosphate (cAMP) accumulation. The cAMP accumulation induced by PACAP-27 and PACAP-38 was five times higher than the cAMP content induced by the other three peptides. The peptide-induced smooth muscle relaxation did not correlate to the cAMP accumulation. NPY (10(-7) M) markedly reversed the relaxations induced by VIP, PHM, PHV, PACAP-27, and PACAP-38, but did not influence the cAMP production induced by these peptides. In conclusion, the relaxation induced by VIP, PHM, PHV, PACAP-27, and PACAP-38 and the contraction induced by NPY are not solely related to the changes of cAMP contents. These findings indicate that in addition to cAMP, another intracellular signal transduction pathway may be involved in the relaxation and contraction induced by these peptides in rabbit ovarian artery.

The influence of neuropeptide Y and norepinephrine on ovulation in the rat ovary.

Neuropeptide Y (NPY) was measured in tissue extracts from ovaries of rats treated with pregnant mare serum gonadotropin (PMSG). The extracted NPY-immunoreactive material was identical to synthetic human NPY with regard to size and hydrophobicity as evaluated by gel filtration and high performance liquid chromatography. The concentration of NPY was related to the estrous cycle and a maximum was observed in relation to the endogenous luteinizing hormone (LH) peak. NPY immunoreactivity was demonstrated by immunohistochemistry to be localized within nerve fibers supplying blood vessels and follicles. The increase in the NPY content could not be related to accumulation around specific ovarian structures. Employing an in vitro set-up, NPY (10(-7) M) was unable to induce ovulation and did not increase the ovulation rate in LH-stimulated ovaries. The combination of NPY (10(-7) M) and NE (10(-7) M) did not significantly increase the number of ovulations compared to that induced by NE (10(-7) M) alone. In conclusion, NPY content in the ovary is related to the estrous cycle, but NPY does not seem to have any direct effect on the ovulatory process.

Pituitary adenylate cyclase activating polypeptide (PACAP): occurrence and vasodilatory effect in the human uteroplacental unit.

UNLABELLED: Pituitary adenylate cyclase activating polypeptide (PACAP) is a neuropeptide which was originally isolated from ovine hypothalamus. PACAP exists in at least two biologically active forms, PACAP-38 and PACAP-27. The aim of this study was to establish the distribution, localization and smooth muscle effects of PACAP-38 and PACAP-27 in the human uteroplacental unit. For this purpose we used radioimmunoassay, immunocytochemistry and in vitro studies of the effect of the peptides on smooth muscle activity. RESULTS: By radioimmunoassay both peptides were detected throughout the uteroplacental unit. The concentrations of PACAP-27 were in general low, ranging from 1/6-1/25 of the corresponding PACAP-38 concentrations. PACAP-immunoreactivity was localized in nerve fibres of the lower segment of the pregnant uterus, but the number of PACAP-immunoreactive nerves was very clearly reduced compared to the corresponding isthmic region of non-pregnant myometrial tissue. PACAP-immunoreactive fibres were not observed in placenta or in the umbilical cord. Both PACAP-38 and PACAP-27 caused a concentration-dependent relaxation on stem villous arteries and on the intramyometrial arteries. Neither of the peptides displayed any effect on non-vascular smooth muscle specimens from the term pregnant myometrium. In conclusion the findings suggest a vasoregulator role of PACAP in the human uteroplacental unit.

Neuropeptide Y in the human prenatal and mature gonads.

In the present paper we have examined the distribution and identity of neuropeptide Y (NPY) immunoreactivity in the prenatal and mature human gonads. Tissue specimens were obtained from prenatal gonads (gestational age < 42 weeks) of induced abortions, premature birth and stillbirths. Specimens were also taken from infants who died aged less than 4 days. Tissue from mature gonads was obtained during surgery. The distribution and concentration in tissue extracts were determined by immunohistochemistry and radioimmunoassay. NPY-immunoreactivity (NPY-IR) was demonstrated in both the prenatal and mature human gonads. In the gonads from fetuses at 16-25 weeks the NPY-IR was scarce and could only be demonstrated by radioimmunoassay. In the female gonads from fetuses at 40 weeks of gestation, distinct nerve fibres were detected in relation to blood vessels and primordial follicles. In the prenatal male gonads at the same gestational length as well as in the mature testis, the NPY-IR nerves were detected in relation to blood vessels and seminiferous tubules. The gel filtration profile from the prenatal gonads and from the mature ovary revealed a single peak corresponding to the elution position of bioactive synthetic human NPY. The gel filtration profile from the mature testis demonstrated an additional peak at Kd of 0.17. The presence of NPY-IR in relation to the developing and mature gonads suggests that NPY participates in gonadal functions from an early point in life.

Structure-function studies on neuropeptide Y and pancreatic polypeptide--evidence for two PP-fold receptors in vas deferens.

The biological effects of neuropeptide Y (NPY), rat pancreatic polypeptide (rPP), hybrid analogs of NPY and PP, and C-terminal fragments of NPY were studied in the field-stimulated rat vas deferens model. The results were correlated with peptide binding experiments in Y1 and PP receptor assays on rat PC-12 cells and Y2 receptors on porcine hippocampal membranes. NPY and rPP inhibited the electrically induced contractions in the vas deferens with an IC50 of 25 and 22 nM respectively. However, in contrast to NPY, rPP could not totally block muscle activity. The inhibitory action of the long C-terminal fragment of NPY, NPY-(19-36) and NPY-(11-36), indicated that NPY acts through a Y2 receptor in the vas deferens. The structural basis for the differential recognition of NPY and PP by Y2 receptors and partly also by PP receptors, could be defined with hybrid analogs of PP and NPY. The analogs, [Ile31,Gln34]PP and [Leu31,Pro33]NPY reacted in the vas deferens preparation in accordance with their relative potency in the Y2 and PP receptor assays. [Ile31,Gln34]PP, which bound to the Y2 receptor like NPY, was also able to block the part of the contractile response which was resistant to rPP. It is concluded that in the vas deferens, PP-fold peptides act through two types of receptors: Y2 and PP, and that residues in the C-terminal part of the molecules determine the differential recognition of the peptides by these receptor types.

Pre-harvest surveillance of Campylobacter and Salmonella in Danish broiler flocks: a 2-year study.

In national surveillance programmes of broiler flocks carried out in Denmark during 1998 and 1999, 89,110 samples for Campylobacter representing 8911 broiler flocks were taken at 10 different abattoirs, and 44,550 samples for Salmonella were taken from the same flocks in the broiler houses at the farms. Of the swabs, 42.5% were Campylobacter positive. Most positive samples were found during July, August and September, while the lowest number of positive samples were found during January, February, March and April. Of the flocks, 5.5% were Salmonella positive, but no seasonal variation was observed. For each flock, the presence of Campylobacter and Salmonella was recorded in order to estimate the possible correlation between colonisation with the two pathogens. In conclusion, no significant effects on intensive cleaning and disinfection procedures on Campylobacter occurrence could be demonstrated, and no significant correlation between occurrence of Campylobacter and Salmonella infections in Danish broilers could be demonstrated which is in contrast to previous observations on concurrent colonisation of broilers with these two zoonotic pathogens.

Pasteurella multocida from outbreaks of avian cholera in wild and captive birds in Denmark.

An outbreak of avian cholera was observed among wild birds in a few localities in Denmark in 2001. The highest mortalities were among breeding eiders (Somateria mollissima) and gulls (Larus spp.). Pulsed-field gel electrophoresis (PFGE) was conducted using ApaI and SmaI as restriction enzymes and restriction enzyme analysis (REA) using HpaII. The Pasteurella multocida subsp. multocida strain isolated from birds in this outbreak was indistinguishable from a strain that caused outbreaks in 1996 and 2003. Most isolates from domestic poultry had other PFGE patterns but some were indistinguishable from the outbreak strain. Among 68 isolates from wild birds, only one PFGE and one REA pattern were demonstrated, whereas among 23 isolates from domestic poultry, 14 different SmaI, 12 different ApaI, and 10 different HpaII patterns were found. The results suggest that a P. multocida strain has survived during several years among wild birds in Denmark.

Verrucous endocarditis associated with Streptococcus bovis in mink (Mustela vison).

Between 1998 and 2001, mortalities due to verrucous endocarditis were experienced at several mink farms. Gram-positive cocci were isolated from the endocardium of all the animals examined but not always from other internal organs. Almost all the isolates were identified as Streptococcus bovis and only a few isolates belonged to other Streptococcus species. Typing by pulsed-field gel electrophoresis of a selection of isolates revealed several patterns and several different clones. Attempts to reproduce disease by the injection of cultures of a field isolate into healthy mink failed.

Serovars of Salmonella isolated from Danish turkeys between 1995 and 2000 and their antimicrobial resistance.

The prevalence of Salmonella serovars and their antimicrobial resistance patterns were investigated among Danish turkeys between 1995 and 2000, by sampling the flocks approximately 14 days before they were slaughtered. Within the flocks, the prevalence of salmonella varied from 7.1 per cent to 25 per cent, and 24 different serovars were detected. The five most prevalent, which accounted for 58.5 per cent of the isolates were Salmonella Heidelberg (16.2 per cent of the isolates), Salmonella Agona (15.8 per cent), Salmonella Derby (12.4 per cent), Salmonella Muenster (7.3 per cent) and Salmonella Anatum (6.8 per cent). In addition, a few rough isolates and isolates belonging to the antigenically incomplete formulae 6,7:-:- and 4,12:b:- were found. The level of antimicrobial resistance was low; the highest resistance was recorded to ampicillin (13.7 per cent) and streptomycin (9.0 per cent) followed by tetracycline (8.5 per cent), sulphonamides (7.7 per cent) and spectinomycin (4.7 per cent). Resistance to quinolones was very low: four isolates were resistant to nalidixic acid, and only one was resistant to enrofloxacin. No resistance was recorded to colistin, apramycin, ceftiofur, florfenicol, or amoxycillin with clavulanic acid. Only 24 isolates were resistant to two or more compounds in various combinations of up to six compounds; one Salmonella Havana isolate was resistant to six compounds. Six isolates were serovar Typhimurium, but none of them belonged to phage type DT104.