Browsing by Author "Bishop, J. Russell"

Now showing 1 - 14 of 14
  • Thumbnail Image
    1991 federal regulations for drug residues in milk
    Jones, Gerald M.; Bishop, J. Russell (Virginia Cooperative Extension, 1992)
    Discusses federal standards and regulations for milk quality and the proper use and storage of animal drugs on dairy farms.
  • Thumbnail Image
    Assessment of dairy product quality by selective incubation and rapid techniques
    Byrne, Robert Duane (Virginia Tech, 1989-12-05)
    A study was conducted to investigate the use of bacterial numbers and metabolites for their Value in predicting potential shelf-life of pasteurized fluid milk. The study consisted of two phases. In the first phase, chemical inhibitors were evaluated for their ability to allow growth of gram-negative psychrotrophic bacteria while inhibiting growth of gram-positive organisms. Crystal violet, benzalkonium chloride, a mixture of crystal violet and benzalkonium chloride, and alkylaryl sulfonate were tested. These were incubated with milk samples separately inoculated with gram-negative and gram-positive bacteria, at 2l°C for 18 hours. Alkylaryl sulfonate allowed growth of gram-positive organisms. Crystal violet inhibited gram-positive organisms but was not as effective as the mixture of crystal violet and benzalkonium chloride, The inhibitor of choice was benzalkonium chloride as it inhibited growth of all gram-positive organisms tested and allowed growth of the gram-negative psychrotrophic organisms.
  • Thumbnail Image
    Attachment of bacteria to teflon and buna-n-rubber gasket materials
    Gaspar-Rolle, Maria Nelma Pinto (Virginia Tech, 1991-02-15)
    Surface analysis of buna-N-rubber and teflon was performed. Scanning electron microscopy was used to analyze the topography of both materials and x-ray microanalysis identified the elemental chemical composition of the polymers. Teflon was primarily a smooth surface with random irregular spots, while buna-N-rubber had a very rough topography with "caverns" and crevices spread over the surface. The x-ray microanalysis showed that there are no impurities on the surface of teflon; however, calcium, silicone and sulfur were present on the surface of buna-N-rubber. Water contact angle measurements indicated that buna-N-rubber was a more hydrophobic surface than teflon. Qualitative analysis of the attachment of Pseudomonas fragi A TCC 4973, Listeria monocytogenes Scott A and Bacillus cereus ATCC 11778 to buna-N-rubber and teflon was assessed by scanning electron microscopy. These bacteria readily attached to both surfaces. Pseudomonas fragi attached after 2 hours in the presence of this microoorganism and Bacillus cereus and Listeria monocytogenes attached at 12 and 24 hours, respectively. Quantitative analysis of the attachment of Pseudomonas fragi to both surfaces as affected by various milk fat concentrations and temperature, and the availability of nutrients (different dilutions of skim milk, casein, casein and lactose, and whey and lactose) was conducted. Attachment was assessed by impedance microbiology. Milk fat content did not play a significant role in the process of attachment of this organism to either type of surfaces; however, significantly greater numbers attached to buna-N-rubber than to teflon. Overall bacteria attached in higher numbers to both surfaces when grown at 21°C, compared to bacteria grown at 4°C. For buna-N-rubber, bacteria attached in significantly higher numbers when the concentration of nutrients was minimal, while for teflon, the results were, in most cases, opposite to these.
  • Thumbnail Image
    Capture filtration for concentration and detection of selected microorganisms in milk
    Byrne, Robert Duane (Virginia Tech, 1994-05-04)
    The effectiveness of an adsorption filter in retaining bacteria present in milk was examined. Skim milk and whole milk (100ml) were separately filtered through a 47mm adsorption filter. No significant change in total solids, total fat, and solids-not-fat percentages of skim and whole milk permeates was observed after filtration. Adsorption of Pseudomonas fluorescens at target concentrations of 103 , 102 , and 101 cells/ml was determined in 100ml of dairy standard methods buffer, nutrient broth, whole milk, and skim milk. The average percentage bacterial retentions were 95 ± 5.5%, 95 ± 2.6%, 28 ± 22.1%, and 62 ± 15.5%, respectively. A treatment was developed for milk to increase the bacterial retention of ~ fluorescens after filtration. The preferred treatment for 100ml of skim milk involved the following final concentrations (v/v): 0.80% disodium ethylenediamine-tetraacetic acid, 0.02% sodium dodecyl sulfate, pH to 7.5 with 1N sodium hydroxide. The average bacterial retention of ~ fluorescens using the treatment was 91 ± 7.1%. Enumeration of bacteria adsorbed to the filter was then conducted using impedance microbiology. When milk was inoculated with ~ fluorescens at target concentrations of 103 , 102 , and 101 cells/ml, an average log bacterial increase of 1.4 ± 0.1 (25x) was obtained. This method will allow for rapid detection of microorganisms in milk by increasing microbial load in the tested sample and eliminating the need for pre-enrichment.
  • Thumbnail Image
    A comparative study of the structure and biochemical activity of flight muscles from several insect species
    Guerra, Antonio Alvarez (Virginia Tech, 1965-02-05)
    Comparative histological and biochemical studies were conducted with the flight muscles and mitochondria of several insect species to determine any correlation which exists between muscle structure and the metabolic activity of their mitochondria. Light microscopic examinations were used in the subcellular observations of these muscles. These included direct observations of freshly isolated material (wet mounts), as well as permanent preparations of transverse sections of some muscle fibers. Histological results were presented in a table which includes the muscle type, as well as the fiber, fibril, and mitochondria sizes of several insect species. The presence and abundance of mitochondria and their arrangement in straight rows between the fibrils, was shown to be common in the flight muscles of all insects investigated (except in the female cockroach, Periplaneta americana). In support of these results, a series of microphotographs showing a comparison of the features studied is presented.
  • Thumbnail Image
    Concentration of sulfamethazine in spray dried milk
    Malik, Shahana (Virginia Tech, 1991)
    A study was conducted to investigate the effect ofspray drying on concentration of sulfamethazine (SMZ) in fluid milk dried to powder «10% moisture). Antibiotic-free skim and whole (homogenized) milk were spiked at 5, 10, 50 and 100 ppb sulfamethazine levels, pasteurized and stored at 4°C till further processed. All samples were spray dried at an inlet temperature of 180 ± 2°C and outlet temperature of 100 ± 2°C and stored at -20°C until analyzed. Sulfamethazine concentration was determined quantitatively by HPLC, a microbial receptor assay (Charm-II®) and an ELISA assay (LacTek®) and qualitatively by an ELISA method (Cite®) in milk samples before and after spray drying. Dry milk samples were reconstituted (10% w/w) for all analyses. statistical determination of significant differences (p = 0.05) between fluid and dry milk samples and whole and skim milk sample~was completed by paired t-tests. Sulfamethazine concentrations increased 81.4% and 84.1% in skim and whole milk respectively at 100 ppb spiked level but were lower than expected increase of 88-91% based on their total solids for whole and skim milk as obtained by modified FDA HPLC method. At lower levels of 5 and 10 ppb, the HPLC method was not sensitive enough to provide usable data. Increase in sulfamethazine concentration from fluid to dry milk was also determined by· Charm-II® and LacTek® techniques. Poor recoveries and variability in data were evident due to binding of sulfamethazine to undetermined milk components as a result of processing and storage also due to break-down of sulfamethazine (mp = 176°C) at 1S00C during spray drying. Sulfamethazine 163 ppb by LacTek® and 94.6 ppb by Charm-II® (at a spiked level of 10 ppb fluid milk) was successfully removed from dried milk after 120 min using supercritical CO₂ (pressure = 5500 psi, 50 0 C
  • Thumbnail Image
    Detection of the fluorescing group of Pseudomonas by enzyme-linked immunosorbent assay (ELISA) for the prediction of shelf-life of dairy products
    Dishart, Katy Johanna (Virginia Tech, 1991-11-05)
    An enzyme-linked immunosorbent assay (ELISA) using polyclonal antibodies has been developed for the detection of the fluorescing group of Pseudomonas. The assay was used as a rapid test (6h) for predicting the shelf-life of pasteurized fluid milk. Milk samples were held at 7°C and tested weekly until determined to be unacceptable by daily sensory evaluation. Sterile milk samples were inoculated with target concentrations of 0 (control), 100, and 1000 cells/ml of Pseudomonas fluorescens on day 0. Samples were tested before and after preliminary incubations. Preliminary incubations conducted include milk alone and milk with broth (1:1) for 18h at 21°C. ELISA and plate counts were performed before and after preliminary incubation to determine the number of pseudomonads present and the relationship between ELISA and plate counts. These numbers were correlated to the shelf-life of each sample, as determined by sensory evaluation. Samples undergoing a preliminary incubation with only milk gave the best correlation to shelf-life (R=0.86).
  • Thumbnail Image
    Development of female sex organs during aestivation in the alfalfa weevil, Hypera postica (Gyll.) (Coleoptera, Curculionidae)
    Guerra, Antonio A. (Virginia Tech, 1962-06-05)
    Laboratory studies and field observations were conducted to ascertain the development of the female sex organs of the alfalfa weevil during aestivation, to study the role of this aestivation on sexual development, and to investigate the factors responsible for this interaction. Measurements of female sex organs were made in dissected weevils at monthly intervals to observe each of the developmental stages of sexual maturation. Results of this sexual development as it progressed after adult emergence, were presented by means of tables, graphs, and a series of photographs. The importance of not confusing old and new weevils was recognized in this work, since their behavioral patters differ in many respects. The nature and physiology of the aestivation of this weevil was discussed and evidence was presented to support the author's suggestion that this aestivation is a "true diapause." It was demonstrated in this paper that diapause synchronized the sexual growth of the alfalfa weevil to the seasonal changes of the year for species adaptation and survival.
  • Thumbnail Image
    Effect of processing parameters on the detection of animal drug residues in milk
    Conner, Tonya Michele (Virginia Tech, 1992-08-05)
    The advent of new methods to detect animal drug residues has resulted in a need to independently validate them. The effects of processing milk on the performance of these methods was evaluated. Antibiotic-free milk samples were spiked with sulfamethazine, penicillin G, and chlortetracycline at levels of 10, 10 and 30 ppb, respectively. Spiked milk and negative control milk was heat-treated, homogenized or heat-treated and homogenized. The procedures evaluated for penicillin detection were Bacillus stearothermophilus disk assay, a HPLC described by Barker et al., Charm II microbial receptor assay, and CITE Probe and LacTek enzyme immunoassays. The procedures evaluated for sulfonamide detection were an HPLC method described by Long et al., Charm II microbial receptor assay, CITE Probe, LacTek and Signal enzyme immunoassays. The methods evaluated for tetracycline detection were a HPLC method described by Long et al., Charm II microbial receptor assay, and LacTek and CITE Probe enzyme immunoassays. The results indicate that the commercial tests and the disk assay were not adversely affected by processing treatments. Significant treatment differences were found when testing raw Charm II data by analysis of variance but these differences did not effect the overall results of the test. Results of the HPLC method were inconclusive for the three drugs tested.
  • Thumbnail Image
    Effects of using an enzyme-linked immunosorbent assay to monitor the control of Staphylococcus aureus mastitis in dairy herds
    Grove, Tina Moler (Virginia Tech, 1990-07-05)
    Bovine mastitis is the most important economic disease to the dairy industry with losses estimated at 2 billion dollars per year in the United States. Staphylococcus aureus (.§.. aureus) is the primary cause of contagious mastitis. Conventional culture methods (National Mastitis Council) were used as a basis for comparing the ability of the enzyme linked immunosorbent assay. ProStaph Iâ ¢, to identify s. aureus. The test had an accuracy of 96%, with a sensitivity of 90% and a specificity of 97%. Results indicated that rinsing teat-cup liners with a 25 ppm iodophor or 100 ppm chlorine solution reduced the presence of S. aureus on the liners by 97%. ProStaph I was used to rapidly screen DHIA preserved milk samples in 10 Virginia cooperator herds. Herds were classified as high (>10% infected) or low prevalence (<10% infected). There were six high prevalence herds after the first test. Average prevalence of cows scoring Ab +2 and +3 was 11.9% ± 7.9. Over the seven month study, prevalence of positive cows declined significantly (P<.OI), but somatic cell count remained relatively unchanged (P>.lO). Four herds continued to have >10% of the animals infected. Incidence of new infection averaged 3.6% ± 2.8 from the first to the last test. Chronic cows averaged 6.9% ± 4.8 over the seven month study. Analysis of variance showed significant (P<.Ol) effects of herd on ProStaph I score J milk yield, and see. Elevated ProStaph I scores were highly correlated (P <.01) with increases in lactation number. ProStaph I changed quadratically (P<.Ol) with increasing SCC. Somatic cell count increased (P<.OI) as ProStaph I score increased.
  • Thumbnail Image
    Growth and survival of Clostridium botulinum type E in pasturized oysters
    Bucknavage, Martin M. (Virginia Tech, 1988-12-09)
    The risk of toxin production by Clostridium botulinum type E in pasteurized oysters was evaluated. Thermal death time studies for type E spores in a oyster homogenate showed that the spores survived pasteurization at 550 C; D-values ranged from 65 to 100 min at 700 C and 880 to 1300 min at 550 C. When tubes of oyster homogenate were inoculated with 1x103 type E spores/ml and stored at 3, 6, 12, and 30oC, no growth was observed; however, when the inocculum was increased to 1x104 spores/ml, toxin was produced during storage at 30oC, but not at lower temperatures. For containers of whole oysters inoculated with a low spore level, toxin production occurred during incubation at 6, 12, and 30oC.
  • Thumbnail Image
    Increase in heat resistance of Listeria monocytogenes Scott A by sublethal heat shock
    Linton, Richard Howard (Virginia Tech, 1991-02-05)
    Log phase cells of Listeria monocytogenes Scott A were heat shocked in Trypticase Soy + 0.6% Yeast Extract broth at 40, 44, and 45°C for 3, 10 and 20 min at each temperature, followed by heating at 55â C for 50 minutes in order to determine an optimum heat shock response. Most heat shock temperatures significantly increased thermal resistance (p < 0.05). Increasing heat shock temperature and time allowed the organism to survive much longer at 50 to 65°C than nonheat shocked cells. The optimal heat shock condition was 45°C for 20 min where D-values at 55°C increased 2.3 fold in non-selective agar and 1.6 fold in selective agar in comparison to non-heat shocked cells. However, cells heat shocked at 48°C for 10 min gave more consistent results; these cells were heated at 50, 55, 60, and 65°C to determine a z-value. Although D-values notably increased due to heat shocking, z-values remained constant regardless of the plating medium. When aerobically heat shocked cells (45°C for 10 min) were plated on a non-selective or a selective medium, a 1.4x increase in D-value was observed when enumerated under strictly anaerobic conditions. Aerobically heat shocked cells (48°C for 10 min) added to shrimp samples retained the increased heat resistance at 55°C when enumerated on a nonselective medium compared to the non-heat shocked cells. Heat shocking conditions may be created in pasteurization or minimal thermal processing of foods allowing increased heat resistance of pathogenic and spoilage microorganisms.
  • Thumbnail Image
    Sanitizer efficacy towards attached bacteria in a simulated milk pipeline system using pure and mixed cultures
    Mosteller, Tracy M. (Virginia Tech, 1993)
    The efficacy of six sanitizers, [chloline (200 ppm), iodophor (2S: ppm), acid anionic (200 ppm), peracetic acid (200 ppm), and fatty acid sanitizer (200 ppm)], was evaluated against bacteria attached to gasket materials. Pseudomonas fluorescens, Yersinia enterocolitica, Bacillus cereus, and Listeria monocvtogenes were capable of significant attachment to both buna-N nlbber and Teflon® gasket surfaces in either pure or mixed cultures. Differences in initial attachment rates were evident in a mixed culture of P. fluorescens, Y. enterocolitica, and Listeria monocytogenes in vitro. Sanitizer effectiveness depended upon the bacterium being enumerated, the type of surface, if the bacterium was attached in pure culture or as part of a mixed culture, and the system of evaluation, (i.e. whether or not sanitizer was used alone or as part of a cleaning system). Peracetic acid was the most effective. Removal of bacteria was more pronounced on the Te'f1on® surface with all sanitizers used. The cleaning system, which consisted of a pre-rinse with warm water, application of the cleaning solution, post-rinse with warm water, and application of the sanitizing solution, allowed microorganisms to remain, when the bacteria were present as a pure culture, but resulted in the complete removal of bacteria in mixed culture.
  • Thumbnail Image
    Use of Antimycotics, Modified Atmospheres, and Packaging to Affect Mold Spoilage in Dairy Products
    Grove, Tina Moler (Virginia Tech, 1998-11-24)
    The effects of natamycin, oxygen scavengers and a 25% CO₂:75% N₂ modified atmosphere on the growth of Penicillium roqueforti in shredded cheddar and mozzarella cheese stored at 10°C for 0, 60, 120, or 180 day was studied. Microbiological and sensory testing was assessed on 0, 7, 14 and 21 days after opening. Carbon dioxide decreased (P<0.05) as O₂ (P>0.05) and N₂ (P>0.05) increased throughout storage. Cheddar and mozzarella cheeses were stored for 180 and 60 days, respectively without significant (P> 0.05) increases in yeast and mold populations. Fungal populations increased significantly (P< 0.05) after packages were opened. Differences in yeast and mold (YM) counts during storage and once the packages were opened were independent of natamycin application and presence, O₂ scavengers and inoculated Penicillium roqueforti for both types of cheeses. Growth of Penicillium roqueforti, Aspergillus niger, Geotrichum candidum and Neosartorya fischeri were evaluated in atmospheres of 0:30:70, 0.5:29.5:70, 1:29:70, 2:28:70, and 5:25:70, O₂:CO₂:N₂ over a 5-day period. Spores were cultured on antibiotic-supplemented potato dextrose agar (pH 5.6, aw 0.95) and incubated at 25°C. All four molds germinated and grew at 0.5:29.5:70. Extent of mycelia growth diameter (mm) increased significantly (P<0.05) as oxygen concentration increased from 0.5% to 5%. All growth was inhibited at 0:30:70, but germination and growth occurred once cultures were exposed to 20.9% atmospheric O₂, indicating that a modified atmosphere containing no residual O₂ is fungistatic. Yeast and mold growth was seen in ultra-pasteurized (UP) extended shelf-life fluid milk stored at (7.2°C). Ten half-pint, pint, quart and half gallon filled cartons were randomly selected from all UP products available. Samples, pulled at random on day 0, 15, 30, 45, and 60, were plated on Yeast and Mold Petrifilm™. Forty-seven percent of the UP products stored for 45 days tested positive for mold. Fungal growth was apparent down the side and along the bottom of the 5th panel. Contamination was traced to the presence of yeast and mold spores in paperboard cartons. Pinholes were present in the polyethylene coating and wicking occurred at the unskived 5th panel. Fungi of similar origin and fatty acid profile were isolated from UP milk products and the paperboard cartons.