Behavior of Listeria monocytogenes in the Presence of Cocoa, Carrageenan, and Sugar in a Milk Medium Incubated With and Without Agitation

1990 ◽  
Vol 53 (1) ◽  
pp. 30-37 ◽  
Author(s):  
LAURA J. PEARSON ◽  
ELMER H. MARTH

Enhanced growth of Listeria monocytogenes strain V7 in chocolate milk rather than skim milk was further investigated by testing various concentrations of cocoa powder (two types of Dutch-process, designated A and B), cane sugar, and sodium carrageenan in skim milk at 13 and 30°C with and without agitated incubation. Increasing sugar concentrations (0, 6.5, and 12.0%) were marginally significant (p = 0.06) in shortening generation times (5.17, 5.07, and 5.05 h, respectively) of the pathogen. Maximum populations attained by the pathogen were greater when cocoa (0.75% type A or B) and sugar (6.5 or 12.0%) were present. Sugar concentration affected growth of L. monocytogenes in an approximately linear relationship (8.41, 8.67, 8.82 log10 CFU/ml for 0, 6.5, and 12.0% sugar, respectively) except in samples containing only carrageenan. In this instance, presence of 6.5 and 12.0% sugar resulted in equivalent maximum populations (8.54 and 8.52 log10 CFU/ml). Three factors enhanced growth of the pathogen at 30°C: addition of cocoa, addition of sugar, and agitated rather than quiescent incubation. Without cocoa, generation times of L. monocytogenes were longer (1.04 h) compared to presence of type A (0.87 h) or B (0.90 h) cocoa. L. monocytogenes in agitated samples had shorter (0.82 h) generation times than in quiescent cultures (0.95 h). Highest populations were attained in agitated samples containing sugar and type A (9.21 log10 CFU/ml) or type B (9.22 log10 CFU/ml) cocoa compared to lowest populations in quiescent samples of skim milk (8.56 log10 CFU/ml).

1987 ◽  
Vol 50 (9) ◽  
pp. 726-729 ◽  
Author(s):  
EILEEN M. ROSENOW ◽  
ELMER H. MARTH

Previously we found that under similar conditions Listeria monocytogenes achieved populations in chocolate milk that were 10 times greater than those in other fluid milk products. The current studies were undertaken to determine why the bacterium grew so well in chocolate milk. Autoclaved samples of milk with 2% milkfat, 2% milk + sugar, 2% milk + cocoa, and 2% milk + sugar + cocoa were inoculated with one of two strains of L. monocytogenes and incubated at 13°C. Carrageenan was also added to one-half of all samples containing cocoa. Growth curves were derived and generation times and maximum populations were calculated for each combination of product and strain of the bacterium. Strain V7 grew faster than strain CA in all products, with most rapid growth occurring in samples containing cocoa (with or without added sugar). Addition of carrageenan further reduced the generation time of this strain. Overall, growth rates ranged from 3 h 55 min (V7 in 2% milk + sugar + cocoa + carrageenan) to 4 h 53 min (CA in 2% milk + cocoa). Product type was primarily responsible for differences in maximum populations achieved by L. monocytogenes. In each instance, final numbers reached were at least 108 cells/ml with highest levels in samples containing all ingredients. The data suggest that sugar, cocoa and carrageenan when added to milk contributed to enhancing growth of L. monocytogenes.


1989 ◽  
Vol 52 (9) ◽  
pp. 625-630 ◽  
Author(s):  
DEMETRIOS K. PAPAGEORGIOU ◽  
ELMER H. MARTH

Autoclaved samples of skim milk and deproteinated whey were fortified with 6 or 12% NaCl, inoculated with Listeria monocytogenes strains Scott A or California (CA), to contain ca. 1.0 × 103 cfu/ml (in the products with 6% salt) or ca. 5.0 × 103 cfu/ml (in the products with 12% salt) and incubated at 4 and 22°C. The pH values of the 6% salted whey, 6% salted skim milk, 12% salted whey, and 12% salted skim milk were 5.65, 6.20, 5.50, and 6.00 respectively. These values remained relatively constant during the entire experiment. Listeria counts were obtained by surface-plating appropriate dilutions and/or undiluted samples on Trypticase Agar (TA). Samples in which L. monocytogenes was not detected, were re-examined after 2, 4, 6 and 8 weeks of cold-enrichment. Generation times of L. monocytogenes in 6% salted whey at 22°C (3.67 h and 3.56 h for strains Scott A and CA, respectively) were significantly shorter than those in 6% salted skim milk at 22°C (4.31 and 4.42 h for the two strains, respectively). Generation times in 6% salted products at 4°C ranged between 37.49 h and 49.43 h. Maximum populations reached at 22 and 4°C ranged from 7.58 to 8.10 Log10 cfu/ml, and were significantly higher in 6% salted whey than in 6% salted skim milk. In 12% salted whey and skim milk incubated at 22°C, L. monocytogenes gradually decreased in numbers. Strain CA was inactivated within 85 d in 12% salted skim milk or within 110 d in 12% salted whey, and was significantly less salt tolerant than strain Scott A which survived for more than 130 d under the same conditions. Loss of viability by both strains was similar in 12% salted whey and skim milk after 130 d of storage at 4°C, and the decreases in population were less than 0.7 order of magnitude.


1987 ◽  
Vol 50 (6) ◽  
pp. 452-459 ◽  
Author(s):  
EILEEN M. ROSENOW ◽  
ELMER H. MARTH

Autoclaved samples of skim, whole, and chocolate milk and of whipping cream were inoculated with Listeria monocytogenes (one to four strains were tested individually, depending on the experiment) and incubated at 4, 8, 13, 21 or 35°C. Growth curves were then derived and generation times and maximum populations calculated for each combination of strain, product, and temperature. The growth rate of L. monocytogenes was similar in all four products at a given incubation temperature and increased with an increase in temperature. Doubling times over all products and strains were 41 min (35°C), 1 h 43 min-1 h 55 min (21°C), 4 h 27 min-6 h 55 min (13°C), 8 h 40 min-14 h 33 min (8°C), and 29 h 44 min-45 h 33 min (4°C). In each instance, maximum populations reached were at least 107 cells/ml, with highest numbers consistently produced in chocolate milk (at least 10 times greater than in skim or whole milk or cream at any temperature). Little decrease in final numbers occurred with extended storage at the incubation temperature being studied. All results were analyzed statistically to determine magnitude and source of variation. Observed differences in data resulted from interactive effects between strain, product, and temperature. Therefore, no single factor can be considered as the sole cause of a particular finding. That L. monocytogenes can attain such high populations at low temperatures should be of concern. Since refrigerated storage is no guarantee of protection against growth of L. monocytogenes, every precaution should be taken to prevent contamination of certain foods by this organism.


1990 ◽  
Vol 53 (1) ◽  
pp. 47-50 ◽  
Author(s):  
LAURA J. PEARSON ◽  
ELMER H. MARTH

Two components of cocoa powder, caffeine (1,3,7-trimethylxanthine) and theobromine (3,7-dimethylxanthine), were evaluated for their effect on growth of L. monocytogenes strain V7. Caffeine (0.5%) and theobromine (2.5%) were added singly or in combination to skim milk or a modified tryptose phosphate broth (MTPB), which were sterilized, inoculated to contain ca. 1 × 103 CFU L. monocytogenes/ml, and incubated at 30°C. Both compounds allowed some growth of the pathogen; however, longer lag phases occurred in samples with (6 to 9 h) rather than without (<3 h) caffeine. Generation times at 30°C ranged from 1.10 h (2.5% theobromine in broth) to 2.28 h (2.5% theobromine plus 0.5% caffeine in milk). Generation times were significantly (p<0.05) lengthened in the presence (2.17 h) rather than in the absence (1.2 h) of caffeine. Populations in samples without caffeine were more than ten times greater (8.57 log10 CFU/ml) than in samples with caffeine (7.21 log10 CFU/ml). Theobromine concentration (0 or 2.5%) and substrate (MTPB or skim milk) had only limited effects on growth of the pathogen. The change in pH of the medium was a function of the extent of growth of L. monocytogenes and the buffering capacity of the substrate. A smaller pH change (0.09 unit) occurred in the milk medium than in broth (0.48 unit) because of the minimal buffering capacity of the 0.2% tryptose solution. Samples with caffeine had a small decrease in pH (0.07 unit) because growth of L. monocytogenes was inhibited and thus acid production was minimal.


1988 ◽  
Vol 51 (4) ◽  
pp. 277-282 ◽  
Author(s):  
DOUGLAS L. MARSHALL ◽  
RONALD H. SCHMIDT

Preliminary studies involving co-inoculation of Listeria monocytogenes with Pseudomonas fragi into whole or skim milk demonstrated that neither inhibition nor stimulation of growth occurred for either organism. Additional investigations involved preincubation of whole milk, skim milk, and 10% reconstituted nonfat dry milk (NDM) for 3 d at 10°C with P. fragi, Pseudomonas fluorescens P26, P. fluorescens T25, or P. fluorescens B52, followed by inoculation with L. monocytogenes and further incubation at 10°C. Growth curves of L. monocytogenes were constructed for each treatment combination and generation times were statistically compared for differences. Results indicated that L. monocytogenes did not affect growth or survival of the preincubated Pseudomonas spp. However, growth rates of L. monocytogenes were significantly (P<0.05) enhanced in milks preincubated with pseudomonads. Doubling times of L. monocytogenes were reduced by up to 3 h when grown in milk preincubated with Pseudomonas spp. Three strains of P. fluorescens showed more stimulation of the growth rate of L. monocytogenes compared to P. fragi in preincubated whole or skim milk but not in preincubated NDM. Milk composition had little effect on growth of either genus when incubated alone. Results of this study indicate that L. monocytogenes can grow in the presence of Pseudomonas spp. either as a co-inoculant or following preincubation in milk at 10°C. Furthermore, data suggest that the presence of the pseudomonads may enhance growth of L. monocytogenes in milk.


1991 ◽  
Vol 54 (5) ◽  
pp. 338-342 ◽  
Author(s):  
FATHY E. EL-GAZZAR ◽  
HANS F. BOHNER ◽  
ELMER H. MARTH

Pasteurized skim milk and retentate (concentrated fivefold or twofold by volume) and permeate from ultrafiltered skim milk were inoculated with Listeria monocytogenes strains California or V7 and incubated at 4, 32, or 40°C. Changes in populations of the pathogen were determined, growth curves were derived, and generation times and maximum populations calculated for each combination of strain, product, and temperature. Both strains grew faster and achieved higher (ca. 1 to 2 orders of magnitude) populations at 4°C in retentate of either concentration than in skim milk. The pathogen grew in permeate at 4°C and attained maximum populations of ca. 106 to 107/ml. Tyndallized samples of skim milk and retentate and permeate from ultrafiltered skim milk were inoculated with the same strains of L. monocytogenes and incubated at 32 or 40°C. Populations achieved by the pathogen at these temperatures, ca. 107 to 108/ml, were similar in skim milk, retentate, and permeate.


Author(s):  
S. Fujinaga ◽  
K. Maruyama ◽  
C.W. Williams ◽  
K. Sekhri ◽  
L. Dmochowski

Yumoto and Dmochowski (Cancer Res.27, 2098 (1967)) reported the presence of mature and immature type C leukemia virus particles in leukemic organs and tissues such as lymph nodes, spleen, thymus, liver, and kidneys of SJL/J strain mice with Hodgki's-like disease or reticulum cell neoplasm (type B). In an attempt to ascertain the possibility that this neoplasia may be of viral origin, experiments with induction and transmission of this neoplasm were carried out using cell-free extracts of leukemic organs from an SJL/J strain mouse with spontaneous disease.It has been possible to induce the disease in low-leukemia BALB/c and C3HZB strain mice and serially transfer the neoplasia by cell-free extracts of leukemic organs of these mice. Histological examination revealed the neoplasia to be of either reticulum cell-type A or type B. Serial transfer is now in its fifth passage. In addition leukemic spleen from another SJL/J strain mouse with spontaneous reticulum cell neoplasm (type A) was set up in tissue culture and is now in its 141st serial passage in vitro. Preliminary results indicate that cell-free material of 39th tissue culture passage can reproduce neoplasia in BALB/c mice.


1992 ◽  
Vol 68 (03) ◽  
pp. 297-300 ◽  
Author(s):  
Monica Galli ◽  
Paul Comfurius ◽  
Tiziano Barbui ◽  
Robert F A Zwaal ◽  
Edouard M Bevers

SummaryPlasmas of 16 patients positive for both IgG anticardiolipin (aCL) antibodies and lupus anticoagulant (LA) antibodies were subjected to adsorption with liposomes containing cardiolipin. In 5 of these plasmas both the anticardiolipin and the anticoagulant activities were co-sedimented with the liposomes in a dose-dependent manner, whereas in the remaining cases only the anticardiolipin activity could be removed by the liposomes, leaving the anticoagulant activity (LA) in the supernatant plasma. aCL antibodies purified from the first 5 plasmas were defined as aCL-type A, while the term aCL-type B was used for antibodies in the other 11 plasmas, from which 2 were selected for this study.Prolongation of the dRVVT was produced by affinity-purified aCL-type A antibodies in plasma of human as well as animal (bovine, rat and goat) origin. aCL-type B antibodies were found to be devoid of anticoagulant activity, while the corresponding supernatants containing LA IgG produced prolongation of the dRVVT only in human plasma.These anticoagulant activities of aCL-type A and of LA IgG's were subsequently evaluated in human plasma depleted of β2-glycoprotein I (β2-GPI), a protein which was previously shown to be essential in the binding of aCL antibodies to anionic phospholipids. Prolongation of the dRVVT by aCL-type A antibodies was abolished using β2-GPI deficient plasma, but could be restored upon addition of β2-GPI. In contrast, LA IgG caused prolongation of the dRVVT irrespective of the presence or absence of β2-GPI.Since β2-GPI binds to negatively-charged phospholipids and impedes the conversion of prothrombin by the factor Xa/Va enzyme complex (Nimpf et al., Biochim Biophys Acta 1986; 884: 142–9), comparison was made of the effect of aCL-type A and aCL-type B antibodies on the rate of thrombin formation in the presence and absence of β2-GPI. This was measured in a system containing highly purified coagulation factors Xa, Va and prothrombin and lipid vesicles composed of 20 mole% phosphatidylserine and 80 mole% phosphatidylcholine. No inhibition on the rate of thrombin formation was observed with both types of aCL antibodies when either β2-GPI or the lipid vesicles were omitted. Addition of β2-GPI to the prothrombinase assay in the presence of lipid vesicles causes a time-dependent inhibition which was not affected by the presence of aCL-type B or non-specific IgG. In contrast, the presence of aCL-type A antibodies dramatically increased the anticoagulant effect of β2-GPI. These data indicate that the anticoagulant activity of aCL-type A antibodies in plasma is mediated by β2-GPI.


2013 ◽  
Vol 1 (1) ◽  
pp. 10
Author(s):  
Noar Muda Satyawan ◽  
Shelly Tutupoho ◽  
Yusli Wardiatno ◽  
Makoto Tsuchiya

Erosion rate on corals due to activities of other biota is called bioerosion. The rock-boring urchin, Echinometra mathaei, when it is abundant, plays a significant role in benthic ecosystems, including biological processes like coral erosion. During feeding, E. mathaei erodes calcium carbonate besides grazing on algae living on coral, so it plays an important role in both organic and inorganic carbons in coral reefs. The urchin E. mathaei actively feeds during the night time (nocturnal grazer). Although in Okinawa four types (A-D) of the urchin exist, the research only focused on the types A and B. Type A of E. mathaei produced 0.44951 g feces per day on average while type B produced 0.38030 g feces per day. CaCO3 analysis in feces and gut contents showed bioerosion rate of E. mathaei type A was 0.64492 g/individu/day, and 0.54436 g/individu/day in type B. There were no significant differences in bioerosion impact of E. mathaei type A and B© Laju erosi pada karang yang disebabkan oleh biota, dikenal dengan bioerosi. Bulu babi jenis Echinometra mathaei, ketika melimpah, menjadi sangat berpengaruh terhadap ekosistem bentik termasuk proses biologi seperti erosi karang. Selama aktivitas makan, E. mathaei menggerus kalsium karbonat dalam proporsi yang besar di samping alga yang tumbuh menempel pada karang sehingga memiliki peran penting dalam siklus karbon organik dan anorganik di ekosistem terumbu karang. Bulu babi E. mathaei aktif mencari makan pada malam hari (nocturnal grazer). Meskipun di Okinanawa ada 4 tipe (A-D), pada eksperimen kali ini memfokuskan pada tipe A dan B saja. Tipe A E. mathaei rata-rata memproduksi 0,44951 g feses/hari dan tipe B memproduksi 0,38030 g feses/hari. Berdasarkan analisis CaCO3 yang dilakukan pada feses dan isi lambung, laju bioerosi yang disebabkan oleh E. mathaei tipe A sebesar 0,64492 g/individu/hari sedangkan tipe B sebesar 0,54436 g/individu/hari. Tidak terdapat perbedaan dampak bioerosi yang signifikan antara E. mathaei tipe A dan B©


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