Total bacterial count and somatic cell count in bulk and individual goat milk around kidding: two longitudinal observational studies.

Total bacterial count (TBC) and somatic cell count (SCC) are important quality parameters in goat milk. Exceeding the bulk milk TBC (BMTBC) thresholds leads to price penalties for Dutch dairy goat farmers. Controlling these milk quality parameters can be challenging, especially around kidding. First, we describe the variation and the peaks around kidding of TBC and SCC in census data on Dutch bulk milk over the last 22 years. Second, to explore causes of these elevations, we studied the variation of TBC and SCC in individual goat milk from 3 weeks before to 5 weeks after kidding and their association with systemic response markers interferon-γ (IFN-γ), calprotectin, ß-hydroxybutyrate (BHB), body condition score (BCS) and fecal consistency. We visited 4 Dutch dairy goat farms weekly for 10 to 16 weeks around kidding. Some of the goats had been dried off, other goats were milked continuously throughout pregnancy. A total of 1,886 milk samples from 141 goats were collected for automated flowcytometric quantification of TBC and SCC measurement. IFN-γ, calprotectin and BHB were determined twice in blood of the same goats, most samples were collected after kidding. The BCS and fecal consistency were scored visually before and after kidding. We found a strong correlation between TBC and SCC (Spearman's rho = 0.87) around kidding. Furthermore, in the third week before kidding, the average TBC (5.67 log10 cfu/mL) and SCC (6.70 log10 cells/mL) were significantly higher compared with the fifth week after kidding, where the average TBC decreased to 4.20 log10 cfu/mL and the average SCC decreased to 5.92 log10 cells/mL. In multivariable linear regression models, farm and stage of lactation were significantly associated with TBC and SCC, but none of the systemic response markers correlated with TBC or SCC. In conclusion, TBC and SCC in dairy goats were high in late lactation and decreased shortly after parturition. For SCC, the dilution effect might have caused the decrease, but this was not plausible for TBC. Moreover, the excretion of bacteria and cells in goat milk was not associated with the selected systemic response markers that were chosen as a read out for general immunity status, intestinal health and metabolic diseases. Therefore, we assume that the TBC increase before kidding and the decrease after parturition is caused by other systemic, possibly hormonal, processes. To reduce BMTBC and BMSCC, it would be advisable to keep milk of goats with highest numbers of bacteria and cells in their milk out of the bulk milk during end lactation. Further studies are needed to investigate the effects of withholding this end lactation milk from the bulk tank.

Phosphorus content of muscle tissue and muscle function in dairy cows fed a phosphorus-deficient diet during the transition period.

Phosphorus (P) deficiency and hypophosphatemia are believed to be associated with muscle function disturbances in dairy cows, particularly around parturition. The objective of this study was to determine the effect of dietary P deprivation during late gestation and early lactation on muscle P homeostasis and muscle function in periparturient dairy cows. Thirty-six multiparous dairy cows in late gestation were randomly assigned either to undergo dietary P depletion or to be offered a diet with adequate P content from 4 wk before to 4 wk after parturition. Phosphorus-deficient rations for dry and lactating cows contained 0.15 and 0.20% P on a dry matter basis, respectively. Blood and muscle tissue for biopsy were obtained and electromyographic examinations were conducted on biceps femoris and intercostal muscles in regular intervals throughout the study. Muscle tissue was analyzed for the total P, adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, creatine phosphate, and tissue water content. Dietary P deprivation resulted in a pronounced and sustained decline of the plasma phosphate concentration, reaching a nadir at calving with mean values below 1.5 mg/dL and remaining below 2.0 mg/dL during the first 4 wk of lactation. Hypophosphatemia was not associated with signs of clinically apparent muscle weakness or disturbed muscle function and was not associated with a decline in the content of any of the studied P-containing compounds in muscle tissue. Accordingly, no association between plasma phosphate concentration and muscle tissue P content was found. Electromyographic examination identified subclinical effects on motor unit action potentials that are indicative of disturbed neuromuscular functionality. Increasing occurrence of pathologic spontaneous activity possibly resulting from membrane instability of nerve or muscle cells and suggestive of myopathy was also recorded as P deprivation progressed. These effects were predominantly observed in intercostal and to a lesser degree biceps femoris muscles. Electromyographic parameters affected by P deprivation were found to be associated primarily with the plasma phosphate and to a lesser extent with the amounts of energy storing P-containing compounds contained in muscle tissue. These results indicate that prolonged and pronounced dietary P deprivation in transition dairy cows leads to marked sustained hypophosphatemia without altering the muscle tissue P homeostasis or causing clinically apparent muscle function disturbances.

The effect of transient, moderate dietary phosphorus deprivation on phosphorus metabolism, muscle content of different phosphorus-containing compounds, and muscle function in dairy cows.

Hypophosphatemia is a common finding in periparturient and anorectic cattle. Although the clinical relevance of hypophosphatemia in cattle is uncertain, it has been empirically associated with persistent recumbency, specifically in periparturient dairy cows. The objective of the present study was to determine if transient dietary phosphorus (P) deprivation over a course of 5 wk, by feeding an approximately 40% P-deficient ration to lactating dairy cows, would result in altered muscle function or muscle P metabolism severe enough to present a risk for animal health and well-being. In addition, we wanted to determine the association between the plasma phosphate concentration ([Pi]) and muscle tissue P content to assess to what extent intracellular P deprivation of muscle cells could be extrapolated from subnormal plasma [Pi]. Ten healthy multiparous, mid-lactating dairy cows received a ration with a P content of 0.18% over a period of 5 wk. Following the P-deprivation phase, the same ration supplemented with P to obtain a dietary P content of 0.43% was fed for 2 wk. Blood and urine samples were collected regularly and muscle biopsies were obtained repeatedly to determine the P content in muscle tissue. Function of skeletal and heart muscles was evaluated by electrocardiography and electromyography conducted repeatedly throughout the study. Feeding the P-deficient ration resulted in the rapid development of marked hypophosphatemia. The lowest plasma [Pi] were measured after 9 d of P depletion and were, on average, 60% below predepletion values. Plasma [Pi] increased thereafter, despite ongoing dietary P depletion. None of the animals developed clinical signs commonly associated with hypophosphatemia or any other health issues. Urine analysis revealed increasing renal calcium, pyridinoline, and hydroxypyridinoline excretion with ongoing P deprivation. Biochemical muscle tissue analysis showed that dietary P depletion and hypophosphatemia were not associated with a decline in muscle tissue P content. Electromyographic examination revealed increased occurrence of pathological spontaneous activity in striated muscles after 2 wk of dietary P depletion in several cows, which could be suggestive of neuromuscular membrane instability. No effect on heart muscle activity was identified electrocardiographically. These results suggest that counter-regulatory mechanisms were sufficient to maintain normal muscle tissue P content during transient and moderate P deprivation. Muscle function was not grossly affected, although the increased occurrence of pathological spontaneous activity suggests that subclinical neuropathy or myopathy, or both, may have occurred with ongoing P deprivation. The results presented here indicate that plasma [Pi] is unsuitable for assessing muscle tissue P content in cattle.

Determination of ceftiofur derivatives in serum, endometrial tissue, and lochia in puerperal dairy cows with fever or acute puerperal metritis after subcutaneous administration of ceftiofur crystalline free acid.

Acute puerperal metritis (APM) is one of the most common diseases during the puerperal period. Systemic administration of ceftiofur for 5 consecutive days has been shown to be effective for treatment of APM. The objective of this study was to determine concentrations of ceftiofur derivatives in serum, endometrial tissue, and lochia of cows with fever postpartum or APM 4 to 6d after treatment with a single subcutaneous dose of 6.6 mg of ceftiofur crystalline free acid (CCFA)/kg of estimated BW at the base of the ear. In the first experiment, samples from CCFA-treated cows with fever postpartum or APM (n=42) were taken on d 4, 5, or 6 after treatment. Concentrations of ceftiofur derivatives were quantified using an HPLC assay. Concentrations of active ceftiofur metabolite desfuroylceftiofuracetamide (DCA) were greatest at d 4 after treatment with CCFA in all samples, but they were considerably lower than the concentrations of DCA in healthy postpartum cows treated with the same dose of CCFA. The concentrations of DCA in serum, endometrial tissue, and lochia were affected by odor of vaginal discharge before treatment with CCFA. Mean concentrations of DCA could be detected above the reported minimal drug concentrations (minimum inhibitory concentrations, MIC) required to inhibit relevant pathogens such as Escherichia coli and Arcanobacterium pyogenes in serum on all days and in endometrial tissue and lochia only on d 4 in CCFA-treated cows with fetid vaginal discharge before treatment. In the second experiment, samples from CCFA-treated cows with APM (n=8) were taken on d 0 (before treatment) and d 4, 5, and 6 after treatment. Mean concentrations of DCA in serum and lochia were similar on d 4 to 6 in both laboratories. Furthermore, determined concentrations of DCA from both laboratories were correlated for serum and lochia. Mean concentrations of DCA could be detected above the reported MIC in serum and lochia only on d 4. Our 2 experiments demonstrated that in postpartum cows with fever postpartum or APM concentrations above the MIC for relevant bacteria (>0.5 µg/mL or >0.5 µg/g) of DCA could be sustained only for 4 (serum: 15/17; endometrial tissue: 2/17; lochia: 1/16) to 5d (serum: 10/13; endometrial tissue: 1/13; lochia: 2/12) after a single treatment with CCFA only in a certain proportion of cows. Overall, our data provide first pharmacological evidence that a single subcutaneous administration of 6.6g of CCFA/kg of BW might not be sufficient to efficaciously treat APM in postpartum dairy cows.

Determination of ceftiofur derivatives in serum, endometrial tissue, and lochia in puerperal dairy cows after subcutaneous administration of ceftiofur crystalline free acid.

Puerperal uterine infections are often associated with decreased reproductive performance in dairy cows. Routine treatment protocols include the systemic administration of antibiotics. Antibiotic drugs, however, should be administered daily over at least 5 d. The objective of this study was to determine concentrations of ceftiofur derivatives in serum, endometrial tissue, and lochia after subcutaneous administration of ceftiofur crystalline free acid in 6 clinically healthy puerperal dairy cows with normal parturition. Samples were taken immediately before treatment, 2 h after, and then every 24 h over a 7-d period. Concentrations of ceftiofur derivatives were quantified using an HPLC assay. In serum and endometrial tissue, ceftiofur derivatives could be detected above the reported minimum drug concentrations required to inhibit relevant pathogens such as Escherichia coli and Arcanobacterium pyogenes over a 7-d period. Concentrations of desfuroylceftiofuracetamide at 5 d after administration of ceftiofur crystalline free acid were 1.21±0.61 µg/mL in serum, 0.86±0.61 µg/mg in endometrial tissue, and 0.96±1.15 µg/mL in lochia. In lochia, mean concentrations of ceftiofur derivatives also remained above the minimal inhibitory concentration of relevant pathogens, but showed greater variations between cows.

Differences in hepatic cytochrome P450 activity correlate with the strain-specific biotransformation of medetomidine in AX/JU and IIIVO/JU inbred rabbits.

Medetomidine is an alpha(2)-adrenoceptor agonist with sedative and analgesic properties. Previously we demonstrated significant differences in the response to medetomidine between two inbred rabbit strains, denoted IIIVO/JU and AX/JU. The aim of the present study was twofold: first, to compare the hepatic CYP450 enzyme activities between these rabbit strains [n = 13(male male,7 female female)/strain]. To this end, liver microsomes were incubated with known fluorescent substrates for the major drug-metabolizing CYP450 isoforms. A comparison of the obtained results indicated significant gender differences as well as differences between the two rabbit inbred strains. Secondly, the biotransformation rate of medetomidine in liver microsomes of both rabbit strains was determined using liquid chromatography coupled to tandem mass spectrometry. The rate of hydroxymedetomidine and medetomidine carboxylic acid formation was found to be significantly higher in the AX/JU strain. Specific CYP2D and CYP2E inhibitors could decrease the formation of both metabolites. Significant correlations were found between the rate of biotransformation of medetomidine and the activities of CYP2D and CYP2E, as well as between CYP450 enzyme activities and the anaesthetic response to medetomidine.

Investigating hydrophilic and electrostatic properties of surfactants using retention on two mixed-mode liquid chromatographic columns.

In environmental risk assessment, it is essential to understand the relationship between molecular structure and fate and toxicity of organic contaminants. For surfactants, physico-chemical parameters which can reflect the interactions that determine surfactant behavior are not well defined and are therefore needed for the development of robust quantitative structure-activity relationships (QSAR). For the present study, we have measured HPLC retention times of several hydrocarbon and perfluorocarbon surfactant groups on a mixed-mode weak anion-exchange (WAX) and mixed-mode hydrophilic interaction liquid chromatography (HILIC) stationary phase. The nonionic alcohol ethoxylates are well retained on the HILIC column. Retention of anionic surfactants on the HILIC column is likely influenced by the degree of hydration of the surfactants and electrostatic repulsion from silanol groups. Less hydrated anionic surfactants (perfluoroalkyl carboxylates, perfluoroalkyl sulfonates and alkyl sulfates) show minimal hydrophilic interaction while other better hydrated anionic surfactants (alkyl carboxylates and alkyl sulfonates) are well retained. The retention mechanism of surfactants on both columns seems to be related to their degree of hydration, albeit expressed in different retention behavior: generally, retention on the WAX phase increases when retention on the HILIC phase decreases, and vice versa. The retention times from both columns were used to calculate retention factors (k') and these were subsequently used in calculating parameters that reflect the electrostatic property (kAX) and hydrophilic property (kHILIC) that determine the interaction between the hydrophilic part of the surfactant and the stationary phase. In further development of predictive models, we suggest the use of kAX for anionic surfactants and kHILIC for nonionic surfactants.

Ceftiofur derivatives in serum, uterine tissues, cotyledons, and lochia after fetal membrane retention.

The objective of the study was to determine concentrations of ceftiofur derivatives after subcutaneous application of ceftiofur hydrochloride in cows with retained fetal membranes. Concentrations of ceftiofur derivatives detected as desfuroylceftiofuracetamide were determined in blood serum, endometrium, caruncles, cotyledons, and lochia during 72 h. After induction of parturition, 2 primiparous and 4 multiparous cows having retained fetal membranes for at least 12 h were studied. All cows received 3 consecutive injections (C1 to C3; 24 h apart) of 1-mg ceftiofur equivalents per kilogram of body weight as ceftiofur hydrochloride sterile suspension. Samples of blood, endometrium, caruncles, cotyledons, and lochia were collected immediately before each injection (0 h) and again at 4, 12, and 24 h after C1, C2, and C3. Blood samples were collected from coccygeal vessels. Caruncles were removed from the uterine lumen by manual extirpation and separated from cotyledons. Endometrial tissue (0.5 g) was collected by using Kenny's biopsy apparatus. For all samples, concentrations of potentially active ceftiofur derivatives were quantified using an HPLC assay. Within 2 h (serum), 4 h (endometrium), and 12 h (caruncles, cotyledons, lochia) after C1 and during the entire study period, mean concentration of ceftiofur derivatives exceeded the reported minimum drug concentrations required to inhibit the growth of 90% of isolates for relevant bacteria such as Escherichia coli, Fusobacterium necrophorum, and Arcanobacterium pyogenes. Only in single samples did concentrations decrease temporarily below the reported minimum drug concentrations required to inhibit the growth of 90% of isolates.

Fluvoxamine maleate: metabolism in man.

The metabolic fate of fluvoxamine maleate in man was investigated. The metabolites were isolated from the pooled urines of healthy volunteers who had ingested either 5 mg radioactive, or 100 mg non-radioactive fluvoxamine maleate as a single dose. The main isolation methods were solvent extraction, column and thin-layer chromatography. Eleven metabolites were isolated; eight of these were carboxylic acids. Identification of nine metabolites was accomplished by mass spectrometry supported by information from the UV spectra and the ionogenic properties. The main route of metabolic degradation of fluvoxamine begins with oxidative elimination of the methoxyl group, another route with removal of the primary amino group. In view of the nature of the degradation pattern none of the metabolites is likely to possess psychotropic activity. For the two primary metabolites this has, in effect, been demonstrated.

Dydrogesterone: metabolism in man.

An investigation of the urinary metabolites of the oral progestational agent dydrogesterone in healthy women of childbearing age is reported. The drug was administered in 3H-labelled form and the urine of the first 8 h, containing on average 38% of the radioactivity administered, was used as the source of the metabolites. It was fortified with urine collected during the first 8 h of a similar study with nonlabelled dydrogesterone. After enzymatic hydrolysis of conjugated metabolites, 43 different chemical species were isolated by means of extraction, followed by column and thin layer chromatography. Three of these metabolites, constituting about 70% of the urinary radioactivity, were positively identified as 20 alpha-hydroxy-9 beta, 10 alpha-pregna-4, 6-diene-3-one (52%), 21-hydroxy-9 beta, 10 alpha-pregna-4, 6-diene-3, 20-dione (18%) and 16 alpha-hydroxy-9 beta, 10 alpha-4, 6-diene-3, 20-dione (1%). Of the remainder, 20 (13%) were tentatively characterized as various products of oxidative attack, all probably having the 4, 6-diene-3-one configuration intact. It is concluded that the 4, 6-diene-3-one configuration is metabolically stable in combination with the 9 beta, 10 alpha configuration. This finding may explain why dydrogesterone, in contrast no progesterone, is orally effective, and lacks estrogenic properties.

Ceftiofur derivates in serum and endometrial tissue after intramuscular administration in healthy mares.

Endometritis is one of the major problems in the horse breeding industry. The use of antibiotics for treatment of endometritis in the mare is recommended as best practice. The intrauterine application of antibiotics, however, has been under discussion over the last years because of concerns about its efficacy. The systemic use of antibiotics has been considered more effective because of its better distribution within the uterus. The objective of the present study was to determine the concentration of ceftiofur derivates in serum and endometrial tissue after intramuscular administration. Specifically, the authors tested the hypothesis that ceftiofur concentrations in serum and endometrial tissue remain above the minimum inhibitory concentration (MIC) for common uterine pathogens for 24 h. Nine mares in estrus received a single dose of 2.2 mg/kg ceftiofur hydrochloride intramuscular per kg of body weight. Blood samples and endometrial tissue were obtained immediately before treatment (-1 h) and 2 h and 24 h after treatment. Endometrial tissue was collected with a Kevorkian biopsy punch. Additional blood samples were collected 4 h and 10 h after treatment from the jugular veins. For determination of ceftiofur derivates in serum and endometrial tissue a high performance liquid chromatography (HPLC) assay was used. Results in serum and uterine tissue revealed greatest concentration of ceftiofur at 2 h and lowest concentrations at 24 h after treatment. Concentrations of ceftiofur at 2 and 24 h after treatment were significantly greater in serum than in endometrial tissue, but remained above the reported MIC for Streptococcus equi zooepidemicus and Escherichia coli in both serum and endometrial tissue until 24 h after treatment.

High-performance liquid chromatographic determination of spectinomycin in swine, calf and chicken plasma.

A rapid clean-up procedure based on ion-pair solid-phase extraction (SPE) for the high-performance liquid chromatographic (HPLC) determination of spectinomycin in swine, calf and chicken plasma at a limit of detection of 50 ng/ml is described. After dilution with water and adjustment of the pH to approximately 5.6, the plasma is applied to a high-hydrophobic C18 SPE column treated with sodium dioctylsulphosuccinate. Spectinomycin is eluted with methanol and derivatized with 2-naphthalene sulphonyl chloride prior to chromatography. The HPLC set-up consists of a dual-column system using two Chromspher silica columns and dichloromethane-acetonitrile-ethyl acetate-acetic acid, in different ratios, as mobile phases. Detection is performed at 250 nm. Quantification is carried out using external standards prepared in blank cleaned plasma. Mean recoveries were 83 +/- 3% (n = 5), 93 +/- 6% (n = 5) and 92 +/- 6% (n = 6) for swine, calf and chicken plasma, respectively, at the 0.1 microgram/ml level.

HPLC determination of residues of spectinomycin in various tissue types from husbandry animals.

An HPLC method was developed for the determination of bacteriostatic aminocyclitol spectinomycin (SP) in animal tissue products. These products included chicken eggs and edible fat, kidney, liver, muscle tissues from calf, poultry, pig and sheep. Residues of SP were extracted from homogenized tissue and egg-derived material with 25 mM citrate of pH 4.0, trichloroacetic acid and dichloromethane. The extract was purified and concentrated over a carboxylic acid-bonded solid-phase extraction (SPE) column. The SPE-eluate was analysed by cation-exchange HPLC involving a two-column switching system, post-column derivatization and fluorescence detection. Spectinomycin could be successfully determined at levels of 0.05 mg kg-1 and higher. Recoveries from spiked tissue material and from spiked egg material were in excess of 74% and did not show a concentration or tissue-type dependence. Precision of the elution position and signal response was better than 2%. Matrix effects and interference from lincomycin were less than 7 and 2%, respectively, on the signal response. Spectinomycin was shown to be stable at -20 degrees C in combined egg yolk and white over a test period of 12 weeks and in calf and sheep muscle tissue over a test period of 10 days. SP was, however, not stable at this temperature over a period of 12 months in chicken muscle tissue. Incurred SP residues were successfully determined in kidney and muscle tissue at the injection site of pigs administered with two doses of 15 mg kg-1 body weight SP with an intermittent withdrawal period of 15 days. Kidney showed higher concentrations and more persistent residues of SP than muscle tissue at the injection site.

High-performance liquid chromatographic determination of spectinomycin in swine, calf and chicken plasma using post-column derivatization.

An HPLC method for the determination of spectinomycin in swine, calf and chicken plasma at 0.1 microgram/ml or higher is described. The clean-up is based upon ion-pair solid-phase extraction on a High Hydrophobic C18 column treated with sodium dioctyl sulfosuccinate. After elution with methanol, spectinomycin is chromatographed on a Spherisorb SCX column using 0.1 M sodium sulphate solution (pH 2.6)-acetonitrile (80:20, v/v) as mobile phase. Fluorescence detection is at an excitation wavelength of 340 nm and an emission wavelength of 460 nm after post-column oxidation with sodium hypochlorite followed by derivatization with o-phthaldialdehyde. Mean recoveries were 99 +/- 2% (n = 6), 99 +/- 2% (n = 7) and 104 +/- 2% (n = 6) for swine, calf and chicken plasma, respectively, at the 0.1 microgram/ml level.

Diflubenzuron: intestinal absorption and metabolism in the rat.

1. The metabolic fate of the insecticide diflubenzuron was investigated in the rat with radioactively labelled forms of the compound. 2. Intestinal absorption, measured as the sum of urinary and biliary excretion, diminished greatly with increasing dose, from about 50% at 4 mg/kg to about 4% at 900 mg/kg. 3. Excretion was almost complete at 72 h after dosing. At that time up to 4% of a dose was recovered from the carcasses of the rats. No detectable excretion of radioactive CO2 occurred (less than 0.5% of dose). 4. The metabolic pattern in urine and bile was investigated with diflubenzuron labelled with both 3H and 14C. No unchanged compound was detected. About 80% of the metabolites appeared to have the basic diflubenzuron structure intact. Three of these, hydroxylated at either aromatic ring, were identified; they were largely excreted as conjugates in the bile. The remainder, also largely excreted in the bile, constituted very polar material. About 20% of the diflubenzuron underwent scission of the ureido bridge. One scission product, 2,6-difluorobenzoic acid, was largely excreted as such in the urine. Its counterpart, 4-chlorophenylurea, was not present in urine or bile in appreciable quantity; nor was 4-chloroaniline detected.

Pharmacokinetics of ceftiofur in plasma and uterine secretions and tissues after subcutaneous postpartum administration in lactating dairy cows.

A study was conducted to measure concentrations of potentially active ceftiofur derivatives, in plasma, in uterine tissues (endometrium and caruncles) and in uterine secretions at different time points after a single subcutaneous administration of ceftiofur hydrochloride (Excenel RTU Sterile Suspension) at the dose of 1 mg/kg body weight in Holstein-Friesian dairy cows. The animals (n=4) were injected within 24 h of calving, after expulsion of the foetal membranes. Plasma, lochial fluid, caruncles and endometrium were collected before ceftiofur hydrochloride administration and at 1, 2, 4, 8, 12 and 24 h after treatment. For each cow the concentrations of ceftiofur in the biological matrices were quantified using an high-performance liquid chromatography (HPLC) assay. The limit of quantification of the method was 0.1 microg/mL for plasma and 0.1 microg/g for lochial fluid, caruncles and endometrium. The concentrations of potentially active ceftiofur derivatives detected in plasma reached a maximum of 2.85 +/- 1.11 microg/mL at 2 h and decreased to 0.64 +/- 0.14 microg/mL at 24 h after administration. In lochial fluid, these concentrations reached a maximum of 0.97 +/- 0.25 microg/g at 4 h and decreased to 0.22 +/- 0.21 microg/g at 24 h after administration. In endometrium, these concentrations reached a maximum of 2.23 +/- 0.82 microg/g at 4 h and decreased to 0.56 +/- 0.14 microg/g at 24 h following the injection, whereas these levels in caruncles were 0.96 +/- 0.45 and 0.60 +/- 0.39 microg/g obtained at 8 and 24 h, respectively. At the dose of 1 mg/kg body weight in healthy dairy cows, subcutaneous administration of ceftiofur (as ceftiofur hydrochloride) after parturition results in concentrations of ceftiofur derivatives in uterine tissues and in lochial fluid that exceed the reported minimal inhibitory concentrations (MICs) for the common pathogens (Escherichia coli, Fusobacterium necrophorum, Bacteroides spp., and Arcanobacterium pyogenes) associated with acute puerperal metritis.

Fluvoxamine: metabolic fate in animals.

The metabolic fate in animals of the antidepressant compound fluvoxamine was investigated. The 14C-labeled drug was administered orally to dogs, rats, hamsters, and mice, and excretion in urine and feces was measured. Chromatographic patterns of the urines were developed by high performance liquid chromatography. These patterns were used as guides in the isolation of the metabolites, its initial step consisting of concentration of the radioactivity in the urine pools in a conical precolumn, followed by separation in the same HPLC system as used for the metabolite patterns. Altogether, 32 radioactive substances were isolated from the urine pools of the four animal species. They were all identified by the combined use of proton nuclear magnetic resonance and mass spectrometry, and by information obtained from chromatographic behavior and color reactions. Several of the 32 compounds were identical, leaving a total of 11 different metabolites in the four species. In all the animal species, the main focus of fluvoxamine degradation was its aliphatic methoxyl group. In three species, this resulted in the corresponding carboxylic acid as the main metabolite, but in the mouse the corresponding alcohol, in glucuronidated form, was at least as important. In mouse and hamster, the methyl ester was a minor metabolite. Products of acetylation or oxidative removal of the primary amino group accounted for only minor proportions of the metabolite patterns. While fluvoxamine itself has the (E)-configuration, several metabolites occurred both in the (E)- and the (Z)-form. The parent compound was isolated only from the urine of dogs, it accounted for less than 10% of the urinary radioactivity.

Proficiency studies on the determination of paralytic shellfish poisoning toxins in shellfish.

Paralytic shellfish poisoning toxins are produced by dinoflagellates. Shellfish filtering these unicellular algae will accumulate the toxins and pose a health risk when consumed by man. In the European Union, paralytic shellfish poisoning toxins in bivalve molluscs are regulated at a maximum content of 80 microg/100 g (91/492/EEC). The current reference method in the European Union is the mouse bioassay, but alternative methods including the liquid chromatography methodology are preferred for ethical reasons. Analyses of suspected shellfish batches revealed, however, unacceptable differences in results reported by a small group of Dutch laboratories all using liquid chromatography methods with precolumn derivatization, followed by fluorescence detection. Therefore, a series of proficiency studies were undertaken among these laboratories. In the first three studies, participants were more or less allowed their own choice of method execution details. This approach yielded unsatisfactory results. A fourth study was then initiated in which a standardized method was mandatory. Two types of test material were used in the fourth study: lyophilized Cardium tuberculatum material containing saxitoxin (STX) and decarbamoyl-saxitoxin (dc-STX), and lyophilized mussel material containing dc-STX. The latter material was investigated in an interlaboratory study involving 15 participants and was considered as the reference material. Among the four laboratories, coefficients of variation (ANOVA) for C. tuberculatum material were 10% (n = 11) and 9% (n = 12) for STX and dc-STX, respectively, and for the reference material was 8% (n = 12) for dc-STX. The joint efforts showed that variability in analysis results between laboratories that all apply more or less the same method can be drastically improved if the methodology is rigorously standardized.