Inhibition of beta-galactosidase biosynthesis in Escherichia coli by tetracycline residues in milk

Low levels of tetracyclines found as residues in milk inhibited the biosynthesis of beta-galactosidase in Escherichia coli. To produce the same effect, other antibacterials had to occur in concentrations that were more than 10-fold higher. This relative selectivity was exploited for the development of a screening test for tetracyclines in milk based on a chemiluminometric assay of beta-galactosidase. The method was validated with spiked samples of raw milk and applied to field samples contaminated with tetracyclines.     

A novel rapid enzyme immunoassay (Fluorophos BetaScreen) for detection of beta-lactam residues in ex-farm raw milk

A novel, qualitative enzyme immunoassay based on fluorescence detection for determination of beta-lactam antibiotics in raw, commingled milk (Fluorophos BetaScreen E. U. test) was evaluated. A dose-response profile for penicillin G was constructed by analysis of spiked milk samples. The limit of detection, defined as the concentration of penicillin G that resulted in 95% of the samples being evaluated as positive, was 1.8 micrograms/kg. The repeatability of the assay was very high both within and between the three participating milk quality testing laboratories. In total 5,061 randomly selected tanker milk samples were analyzed with the BetaScreen test and compared with the Delvotest SP. The agreement between the two tests was 99.7%. Probably due to a higher sensitivity to penicillin G, the BetaScreen test detected almost twice as many suspect positive tanker milk samples (0.45%) as the Delvotest SP (0.26%).     

Confirmation of multiple tetracycline residues in milk and oxytetracycline in shrimp by liquid chromatography-particle beam mass spectrometry

A confirmation procedure is described for detection of residues of six tetracyclines in bovine milk, and oxytetracycline in shrimp. Residues are extracted from milk or shrimp tissue using metal chelate affinity chromatography. The extracts are desalted, further concentrated using polymeric solid-phase extraction, and chromatographed on a polymeric reversed-phase column. Analysis is by methane negative ion chemical ionization on a quadrupole mass spectrometer using a particle beam interface. Data are acquired in partial scan mode, monitoring from m/z 378 to m/z 480. The procedure was validated with control milk and shrimp, fortified milk (30 ng/ml) and shrimp (100 ng/g), and milk and tissue from animals treated with the drugs.     

Gamma-glutamyltransferase as a marker for the pasteurization of raw milk

The degree and rate of inactivation of gamma-glutamyltransferase in raw cow's milk by heating at 50, 60, 70, and 80 degrees C for 1, 2, 3, 5, 10, 15, 20, 25, and 30 min were measured to evaluate the suitability of this enzyme as a marker for the pasteurization of milk. The enzymes alkaline phosphatase and lactate dehydrogenase were also measured under similar conditions for comparison. The patterns of heat inactivation of gamma-glutamyltransferase and alkaline phosphatase were similar, with only a minimal inactivation of the enzymes at 50 degrees C. The rate of inactivation increased as a result of increasing temperatures and time. A complete inactivation of both enzymes was seen at 70 degrees C after 10 min and at 80 degrees C after 1 min. Lactate dehydrogenase showed a higher heat resistance with almost complete inactivation at 70 degrees C for 30 min, and compete inactivation at 80 degrees C for 3 min. No activities of these enzymes were found in commercially pasteurized or heat-treated milk. The levels of gamma-glutamyltransferase in raw milk were between 8 and 10% higher than those of alkaline phosphatase and lactate dehydrogenase, making it more sensitive and accurate as a testing marker. It seems that gamma-glutamyltransferase may serve as a good pasteurization marker. Furthermore, the simplicity of testing and the availability of commercial kits for testing by both wet and dry chemistry make it an attractive choice, especially because dry chemistry procedures overcome the difficulties originating from the turbidity of milk, which interferes with spectrophotometric procedures.     

Determination of tetracycline residues in animal tissues by liquid chromatography

BACKGROUND: Sufficient intraluminal concentrations of 5-aminosalicylic acid (ASA) within inflamed regions of the intestine are required for therapeutic efficacy in inflammatory bowel disease. Various oral delayed release preparations have been developed to ensure that 5-ASA is set free in those parts of the gut, which are most frequently affected. However, resulting intraluminal concentrations within the small bowel are unknown. Therefore, we determined and compared 5-ASA release within different segments of the small bowel from an Eudragit L coated 5-ASA preparation (Salofalk) and from an ethylcellulose coated microsphere preparation (Pentasa). METHODS: Twelve healthy subjects were intubated with an oro-ileal multilumen-tube for marker perfusion, duodenal, jejunal and ileal aspiration of chyme and intestinal manometry. Each subject received 500 mg 5-ASA (Salofalk, n = 6, or Pentasa, n = 6) together with a semiliquid test meal. Intestinal aspirates, blood and urine samples were obtained in regular intervals for 7 to 10 hours and were analysed for 5-ASA and its main metabolite acetyl-5-ASA by HPLC. RESULTS: With Salofalk, gastric emptying of 5-ASA did not take place in the digestive, but in the subsequent interdigestive period. Luminal delivery of 5-ASA and acetyl-5-ASA increased from the duodenum (3% of dose) to the ileum (30% of dose). 10% of the dose administered were excreted in urine and about 90% reached the colon unreleased or solubilised. By contrast, with Pentasa, 5-ASA was delivered to the duodenum together with the test meal and released continuously throughout the small intestine (about 20% of dose solubilised at each intestinal site). Only 3.5% of the dose administered were excreted in urine. Deliver of 5-ASA to the colon was equal to Salofalk. CONCLUSIONS: From both preparations, considerable amounts of 5-ASA are released during small intestinal transit thus explaining therapeutic efficacy in small intestinal Crohn's disease. Because of specific release patterns, Salofalk may be of use especially in terminal ileal disease, where as patients with extensive small intestinal disease including the proximal small intestine might benefit from Pentasa.     

Detection of Listeria in raw and pasteurized milk

We report the case of a 16-year-old adolescent with the onset of a panic disorder with agoraphobia after a first panic attack during marijuana intoxication. There was a good response to standard cognitive behavioural therapy for panic disorder.     

The influence of coffee with milk and tea with milk on the bioavailability of tetracycline

BACKGROUND: At the initial stages of percutaneous transluminal coronary angioplasty (PTCA), several studies reported on the feasibility of coronary artery incision and dilatation leading to the extension of the PTCA technique. HYPOTHESIS: This study was designed to determine the immediate and chronic results of cutting balloon (CB) angioplasty. METHODS: This procedure was performed on 127 lesions in 110 patients (male 83%, age 61.8 +/- 9.3 years). RESULTS: The overall procedural success rates for the CB were 93.7% (119 lesions) and 92.7% (102 patients), while solitary CB without pre- and/or postdilatation was 76.4% (91 lesions). There was one major in-hospital complication (Q-wave myocardial infarction, 0.9%), but there were no deaths or emergency coronary artery bypass graftings. Significant angiographic dissections (> or = grade C) occurred in four patients, and coronary perforation occurred in one. The successfully treated CB group (95 lesions) was matched with the successful conventional angioplasty group (PTCA group) for chronic result assessment in regard to reference vessel size and lesion characteristics. In the CB group, postprocedural minimal luminal diameters were significantly larger and the percentage of stenosis at the stenotic site was significantly lower compared with the PTCA group. Restenosis occurred in 22 lesions (23.1%). This showed a significantly lower restenosis rate compared with the PTCA group (42.1%). In addition, the restenosis rate of the CB without inclusion of the pre- and/or postdilatation-treated lesions was 19.7%. CONCLUSIONS: (1) Cutting balloon angioplasty procedures can be performed with high success rates with few major inhospital events. (2) The restenosis rate in the CB group was significantly lower compared with the PTCA group.     

Rapid enzymatic-colorimetric method for detecting organophosphorous insecticide residues in milk

The authors developed a new enzymatic-colorimetric rapid method for testing milk for residues of organophosphorous insecticides. The test is based on the colour reaction of indoxyl acetate with the esterase of the OROSERAN (Dessau) horse serum which is inhibited by organophosphorous insecticides. Calibration curves were established for Trichlorphon and Dichlorphos (DDVP). The limit of detection lies at 0.02 mg/kg. The time actually needed per sample is about 15 min. The result is obtained within 90 min. Since this procedure requires no clean-up operations, it is labour-saving compared to other thin-layer or gas chromatographic methods. It may be recommended as a screening test for the detection of organophosphorous insecticides in milk samples.     

Effect of psychrotrophic bacteria from raw milk on milk proteins and stability of milk proteins to ultrahigh temperature treatment

The effects of psychrotroph growth in raw milk on proteins of mils and on the response of milk proteins to heat treatments with ultrahigh temperature were studied. Ten gram-negative psychrotrophs isolated from raw milk readily attacked raw milk proteins. Kappa- and beta-casein were most susceptible although some of the isolates also attacked the whey proteins. Detectable proteolysis did not require large psychrotroph populations. A 10 to 20% decrease in kappa-casein during 2 days at 5 C accompanied growth of one isolate to a population of only 10,000/ml. Growth of psychrotrophs in raw milk predisposed the proteins to deleterious effects of ultrahigh temperature treatments. Ultrahigh temperature treatment by direct steam injection had little effect on raw milk caseins and decreased alpha-lactalbumin and beta-lactoglobulin by 21% and 34%, respectively. Milk that had undergone proteolysis exhibited decreased detectable kappa-, beta-, and alphas-caseins and increased loss of beta-lactoglobulin as a result of ultrahigh temperature treatment. Milk suffering extensive kappa-casein degradation coagulated during ultrahigh temperature treatment. Coagulation during or shortly after heating increased with severity of heat treatment and size of psychrotroph population.     

Iodine residues in milk following Iodonal disinfection in the primary production of milk

An isotopic indicator was used for the determination of the presence of iodine residues in milk after the post-milking dipping of teats in Iodonal A. The residues are below the permitted tolerance limit after the cleaning and disinfection of the milking equipment. As to teat dipping in Iodonal A, the residues are high after this treatment and the concentration of the chemical should be decreased from 33% to 20%.     

Organochlorine pesticide residues in cow''s milk in Uganda

Blending the art and science of medicine in the ICU setting is a growing challenge, given modern technological advances, spiraling healthcare costs, and the need to carefully consider evidence from the biomedical literature in our practice. Rather than keeping up to date with all primary research relevant to critical care, intensivists are increasingly seeking synthetic research to help with their day-to-day decision-making. Synthetic research includes systematic reviews, practice guidelines, and economic evaluations. These publications can address questions of prevention, diagnosis, treatment, rehabilitation, and/or palliation, thereby helping intensivists to integrate the results of sound research at the bedside.     

Assay for cephapirin and ampicillin in raw milk by high-performance liquid chromatography--integrated pulsed amperometric detection

The FDA has issued guidelines governing the use of antibiotics in cattle and routinely tests for the presence of antibiotics in milk. Unfortunately, these compounds are often difficult to detect by direct methods because they often lack a chromophore or fluorophore. Integrated pulsed amperometric detection (IPAD) following reversed-phase liquid chromatography is well-suited for this analysis because it is selective, sensitive, and direct; i.e., derivatization is not required. This work involves the development of a simple, rapid assay for the determination of beta-lactam antibiotic residues in milk using HPLC-IPAD, specifically, ampicillin and cephapirin. Since the analyst studied here are detectable by UV detection, a comparison between IPAD and UV detection will be made. Sample preparation schemes that involve the extraction of antibiotics of interest from the milk matrix and subsequent cleanup are an important aspect of this project. These procedures will be discussed in detail. In addition, analytical figures of merit and IPAD wave form optimization will be addressed.     

Organochlorine pesticide residues in cow''s milk and butter in Mexico

In the chipmunk, a mammalian hibernator, a 140 kDa protein complex found in the blood, drastically decreases in concentration during hibernation. This complex contains four species of proteins, HP-20, -25, -27 and -55. In the present study, cDNA clones coding for the chipmunk HP-55 were isolated from a liver cDNA library. Sequence analysis revealed that HP-55 is produced as a precursor protein of 413 amino acids (aa), that it has a signal peptide of 24 aa, and that it contains four potential N-glycosylation sites. The deduced aa sequence shows 63% identity with that of rat alpha1-antitrypsin (alpha1-AT); however, the sequence corresponding to the reactive center P1-P1' residues was found to be Met-Leu, whereas it is Met-Ser in the rat alpha1-AT. During screening of the chipmunk liver cDNA library, four other related classes of cDNA clones were obtained, each also coding for an alpha1-AT-like protein. In spite of more than 86% overall aa sequence identity among the five chipmunk alpha1-AT-like proteins, they are highly divergent in the putative reactive center region; the putative P1-P1' sequences are Met-Leu (HP-55 or CM55-ML), Met-Met (CM55-MM), Met-Ser (CM55-MS), Ser-Ile (CM55-SI) and Ser-Thr (CM55-ST). Each of the alpha1-AT-like protein mRNAs was expressed in chipmunk liver, and the HP-55 mRNA level was greatly reduced during hibernation. Genomic Southern blot analysis and screening of a liver cDNA library from another hibernating squirrel species, the ground squirrel, also revealed expression of multiple members of the alpha1-AT gene family, whereas analysis of a cDNA library from a non-hibernating species, the tree squirrel, found only a single alpha1-AT gene.     

Selective extraction of pyrethroid pesticide residues from milk by solid-matrix dispersion

Fifty human temporal bones from necropsies were used to study the frequency of canal dehiscences in detail along the course of the facial nerve. Specifically, the study focused on bony dehiscences in the fallopian canal and vascular communications between the facial nerve and the surrounding bone. High frequency of dehiscences at the oval window (60%) and in the pyramidal segment (54%) were found. These dehiscence rates are in agreement with published reports. A 20% rate of dehiscences at the most anterior segment of the tympanic segment was noted and a non-reported high rate (20%) of multiple dehiscences along the course of the fallopian canal in the same temporal bone in specimens of newborns and young children. The significance of these findings in terms of clinical implications is discussed.     

Levels of organochlorine pesticides residues in milk of urban mothers in Kenya

Organochlorine compounds (OCPs) are toxic products capable of producing serious adverse health consequences. When used to control pests, certain OCPs persist in the environment and accumulate in the fatty tissues of living organisms, reaching higher levels in animals higher in the food chain. Many countries have therefore either restricted or banned the use of OCPs. DDT used to be widely used in Kenya to control pests, but is now used only to control mosquito populations. The contamination of human milk by organochlorine and other related compounds is a major concern in many parts of the world. Findings are reported from a study conducted to assess the levels of OCPs in an urban area of Kenya and to compare OCP levels in human milk from a previous study in rural areas of the country. 216 milk samples were collected from mothers aged 18-30 years in hospital maternity wards or attending postnatal clinics in selected areas in Nairobi in 1991. All mothers involved in the study were nursing either their first or second child, and had lived in Nairobi for at least 5 years before the date of sampling. Samples were collected from mothers who were up to 4 weeks postpartum and analyzed for the presence of OCP residues. 9 OCPs were detected, with p,p'DDT and p,p'DDE being the most frequently encountered contaminants in all samples analyzed. p,p'DDE was detected in 99.5% and p,p'DDT in 78.2% of all samples. Other residues detected were dieldrin (27%), beta-HCH (18.5%), lindane (12%), and alpha-HCH (8.8%). Levels of sum DDT ranged from 0.004 to 6.321 mg/kg fat. Mothers living in Nairobi have low levels of OCPs in their breast milk compared to mothers in rural areas. The relatively lower urban levels could be attributed to the restricted use of OCPs. People need to be taught how to safely use pesticides.     

Sources and routes of contamination of raw milk with Listeria monocytogenes and its control

To identify the source of contamination of raw bulk milk with Listeria, we attempted to isolate the bacteria from various samples such as cattle-related samples, bulk storage tanks and the environments on three farms. On farms A and B, Listeria monocytogenes was repeatedly isolated from raw milk, while on farm C, it was scarcely isolated from it. On the former farms, Listeria was detected in cattle-related samples and the environments. On the other hand, only one fecal sample on the latter farm was Listeria-positive. Especially, we demonstrated that the bulk tank on farm A was contaminated with L. monocytogenes. Then, L. monocytogenes was controlled by continuously washing the bulk tank on farm A with alkaline detergent.     

Incidence of toxigenic Escherichia coli in soft cheese made with raw or pasteurized milk

The role of cellular Na+ accumulation in acetylcholine-induced desensitization was investigated in guinea pig ileal longitudinal muscle. Desensitization was induced by the pretreatment with acetylcholine (10(-4) M, 30 min) and was expressed by the rightward shift in the concentration-response curve for acetylcholine after the treatment. The same treatment with acetylcholine caused accumulation of cellular Na+ that amounted to about 3.5-fold of the control level. To study the relationship between the gain of cellular Na+ and the development of desensitization, we treated the muscle strips with acetylcholine under the condition in which the external Na+ concentration ranged from zero to 149.2 mM. The result showed that cellular Na+ content is closely related to the extent of desensitization; that is, desensitization was at the lowest level when acetylcholine induced no increase in cellular Na+, while desensitization developed in proportion to the increase in cellular Na+ content. However, when cellular Na+ was increased by another method (by the treatment with ouabain), the inhibition of the acetylcholine response was far less than that observed in the case of desensitization. We concluded that both muscarinic stimulation and the accompanying accumulation of cellular Na+ are required for desensitization to occur in full. This desensitization could be the result of a muscarinic stimulated and cellular Na+-dependent mechanism.     

Prevalence of Listeria species in raw milk from farm bulk tanks in Nagano prefecture

The redox chemistry of Pyrococcus furiosus rubredoxin and ferredoxin has been studied as a function of temperature in direct voltammetry and in EPR monitored bulk titrations. The Ems of both proteins, measured with direct voltammetry, have a normal (linear) temperature dependence and show no pH dependence. EPR monitoring is not a reliable method to determine the temperature dependence of the Em: upon rapid freezing the proteins take their conformation corresponding to the freezing point of the solution.     

Quantitative risk assessment of human listeriosis from consumption of soft cheese made from raw milk

Microbial hazards have been identified in soft cheese made from raw milk. Quantification of the resulting risk for public health was attempted within the frame of the Codex Alimentarius Commission, 1995 approach to quantitative risk assessment, using Monte Carlo simulation software. Quantitative data could only be found for Listeria monocytogenes. The complete process of cheese making was modeled, from milking to consumption. Using data published on the different sources of milk contamination (environment and mastitis) and bacterial growth, distributions were assumed for parameters of the model. Equations of Farber, J.M., Ross, W.H., Harwing, J. (1996) for general and at-risk populations were used to link the ingested dose of L. monocytogenes to the occurrence of listeriosis. The probability of milk contamination was estimated to be 67% with concentration ranging from 0 to 33 CFU ml-1. The percentage of cheese with a predicted concentration of L. monocytogenes greater than 100 CFU g-1 was low (1.4%). The probability of consuming a contaminated cheese serving was 65.3%. Individual annual cumulative risk of listeriosis, in a population each consuming 50 servings of 31 g, ranged from 1.97 x 10(-9) to 6.4 x 10(-8) in a low-risk sub-population and 1.04 10(-6) to 7.19 10(-5) in a high-risk sub-population. The average number of expected cases of listeriosis per year was 57 for a high-risk sub-population and one for a low-risk healthy sub-population. When the frequency of environmental milk contamination was reduced in the model and L. monocytogenes mastitis was eliminated, the expected incidence of listeriosis decreased substantially; the average number of expected cases was reduced by a factor of 5. Thus the usefulness of simulation to demonstrate the efficiency of various management options could be demonstrated, even if results should be interpreted with care (as many assumptions had to be made on data and their distributions.