Melissa C. Newman

Survival of Escherichia coli O157:H7 on Strawberry Fruit and Reduction of the Pathogen Population by Chemical Agents

Survival of Escherichia coli O157:H7 was studied on strawberry, a fruit that is not usually washed during production, harvest, or postharvest handling. Two strains of the bacteria were tested separately on the fruit surface or injected into the fruit. Both strains of E. coli O157:H7 survived externally and internally at 23 degrees C for 24 h and at 10, 5, and -20 degrees C for 3 days. The largest reduction in bacterial population occurred at -20 degrees C and on the fruit surface during refrigeration. In all experiments, the bacteria inside the fruit either survived as well as or better than bacteria on the surface, and ATCC 43895 frequently exhibited greater survival than did ATCC 35150. Two strains of E. coli also survived at 23 degrees C on the surface and particularly inside strawberry fruit. Chemical agents in aqueous solution comprising NaOCl (100 and 200 ppm), Tween 80 (100 and 200 ppm), acetic acid (2 and 5%), Na3PO4 (2 and 5%), and H2O2 (1 and 3%) were studied for their effects on reduction of surface-inoculated (10(8) CFU/ml) E. coli O157:H7 populations on strawberry fruit. Dipping the inoculated fruit in water alone reduced the pathogen population about 0.8 log unit. None of the compounds with the exception of H2O2 exhibited more than a 2-log CFU/g reduction of the bacteria on the fruit surface. Three percent H202, the most effective chemical treatment, reduced the bacterial population on strawberries by about 2.2 log CFU/g.

Immune Parameters of Dry Cows Fed Mannan Oligosaccharide and Subsequent Transfer of Immunity to Calves

Abstract The objective of this study was to evaluate the benefits of supplementation of mannan oligosaccharide (MOS) to cows during the last 3 wk of the dry period on immune function of the cows and subsequent transfer of passive immunity to their calves. Indicators of nonspecific and specific immunity were evaluated. Cows were vaccinated against rotavirus at 4 and 2 wk before expected parturition. Blood samples were obtained from cows before vaccination and at weekly intervals until calving and from calves at birth and 24h for analysis of serum protein concentrations, packed cell volume, white blood cell counts, white blood cell differentials, and serum rotavirus neutralization titers. Colostrum quantity and quality were measured at calving, and immunoglobulin isotype concentrations in colostrum were determined. Specific immunity was enhanced by MOS supplementation as evidenced by greater serum rotavirus neutralization titers at calving in cows supplemented with MOS compared with control cows. Colostral rotavirus neutralization titers were not affected by treatment. Although numerical differences appeared large, there was a high degree of variability in the colostral rotavirus neutralization titers. Calves from cows fed MOS tended to have greater serum rotavirus neutralization titers compared with calves from cows fed the control diet. There was a tendency for greater increases in serum protein concentrations from birth to 24h in calves from cows fed MOS compared with calves from cows fed the control diet. Results indicate that supplementation of MOS to cows during the dry period enhanced their immune response to rotavirus and tended to enhance the subsequent transfer of rotavirus antibodies to calves.

Production of the long-chain alcohols octanol, decanol, and dodecanol by Escherichia coli

As a follow-up to earlier studies on the emission of long-chain alcohols from broth cultures of Gram-negative enteric bacteria, E. coli was examined for the production of 1-octanol, 1-decanol, and 1-dodecanol. Ten strains of E. coli cultured in tryptic soy broth were assayed for volatile metabolites using solid-phase microextraction. Long-chain alcohols were produced by all strains with 1-decanol predominating with production ranging from 23.6 ng mL−1 to 148 ng mL−1. The production of long-chain alcohols followed the onset of the exponential growth phase of the broth culture. Doubling the concentration of glucose (5 g L−1) in the broth had no effect on the concentration of long-chain alcohols produced. Addition of octanoic, decanoic, or dodecanoic acids (as K+ salts) to the broth (100 mg L−1) markedly increased the production of the corresponding alcohols by E. coli, ranging from a 13-fold increase for decanol to a 51-fold increase for dodecanol. However, decanol remained the predominant alcohol detected in all assays. These neutral volatile alcohols may have application as vapor-phase indicators for certain classes of bacteria, particularly, Gram-negative enteric bacteria.

Comparison of long-chain alcohols and other volatile compounds emitted from food-borne and related Gram positive and Gram negative bacteria.

Numerous reports have been published on the antimicrobial activity of synthetic volatile long chain alcohols, such as 1‐decanol and 1‐dodecanol, against bacteria and fungi. The objective of the present study was to survey microorganisms for emission patterns of naturally occurring long chain alcohols and other volatile components to determine if these compounds are associated with certain groups of bacteria. Cultures were grown in trypticase soy broth overnight and volatile compounds were trapped on a porous polymer and identified by mass spectrometry. Subsequently, volatile compounds were collected from 26 strains of food associated bacteria using solid‐phase microextraction and analyzed by gas chromatography. Alcohols comprising 1‐octanol, 1‐decanol, and 1‐dodecanol occurred as products from enteric Gram negative bacteria, which included Citrobacter, Enterobacter, Klebsiella, Salmonella, and Shigella. However, the long chain alcohols were not detected as products from the nonenteric Gram negative species studied which included Acinetobacter, Pseudomonas, and Shewanella. Among Gram positive bacteria, including Bacillus, Enterococcus, Lactococcus, Leuconostoc, Listeria, Staphylococcus, and Streptococcus, the only long chain alcohol detected was 1‐decanol and, if present, it occurred in relatively small amounts. Other classes of compounds emitted by bacteria included methylketones and sulfides. The methylketones were found as products from Gram positive and Gram negative bacteria, whereas the sulfides were closely associated with Gram positive bacteria. In summary, the emission patterns of volatile compounds from bacteria showed many trends including the association of long chain alcohols with enteric Gram negative bacteria. The results provide a basis for future in vivo studies to determine if volatile compounds such as natural long chain alcohols function in the ecology of food‐borne Gram negative bacterial pathogens.

Staphylococcus aureus survival, staphylococcal enterotoxin production and shelf stability of country-cured hams manufactured under different processing procedures

Fresh hams were inoculated with Staphylococcus aureus, cured, equalized, cold smoked or non-smoked, and aged. Initial S. aureus populations of 8.57 and 8.12 Log(10) CFU/cm(2) for salt and salt+NO(2)hams decreased to below the levels of detection after the fourth month of aging. S. aureus was detected following enrichment for 75% of the inoculated and 62% of the control hams at the end of the aging period. Staphylococcal enterotoxin was detected in 40% of the inoculated and 50% of the control hams following the aging period. The NaCl content of these hams with or without nitrite were 4.45/3.37% and a(w) values 0.94/0.91. Country-cured ham products obtained from retail stores in Kentucky were all negative for S. aureus enterotoxin. These results indicate that higher salt content and lower a(w) values on country-cured hams play an important role in controlling the growth and toxin production of S. aureus.

BACTERIA IN THE REPRODUCTIVE TRACTS OF RED-WINGED BLACKBIRDS

Abstract We identified bacteria collected from the ejaculates and cloacae of male Red-winged Blackbirds (Agelaius phoeniceus) and from the cloacae of their mates during two breeding seasons and at three geographic locations with two distinct breeding habitats in each. In all, 42% of 233 ejaculates contained at least one bacteria species. Analysis of control samples suggested that some bacteria come from contamination during collection, but most are either carried within the ejaculate or are present on the male cloaca. As in an earlier study of Agelaius phoeniceus, we recorded many different species with considerable diversity between samples. We tested several explanations for this diversity. We found no overlap in either the presence of bacteria or the species found in multiple ejaculates obtained from the same male. We also found no association between bacteria in the ejaculate and those in the cloaca from the same male, or in samples collected from a male and those from the females nesting on his territory. Surveys of ejaculates from three locations, Kentucky, New York, and Wisconsin, yielded similar patterns of prevalence and diversity. Ejaculate samples from males inhabiting relatively dry upland habitats contained a similar number of bacteria and array of species as males inhabiting relatively wet, marshy territories. Many of the isolated bacteria are probably benign, but pathological effects of some species have been recorded in some circumstances. Our results suggest that acquiring pathogenic bacteria represents a potential cost to multiple mating by females, but the potential benefits of some microbes may offset these risks. Bacterias en el Sistema Reproductivo de Agelaius phoeniceus Resumen. Identificamos bacterias aisladas del semen eyaculado y de las cloacas de machos de la especie Agelaius phoeniceus y de las cloacas de sus parejas hembras durante dos estaciones y en tres áreas geográficas, con dos tipos de hábitat en cada área. En general, el 42% de las 233 muestras de eyaculaciones presentó por lo menos una especie de bacteria. El análisis de las muestras control sugiere que algunas bacterias provienen de contaminación ambiental durante la colecta, pero la mayoría de las bacterias provienen del semen o están presentes en la cloaca del macho. Al igual que en un estudio anterior con Agelaius phoeniceus, registramos muchas especies distintas, con bastante diversidad entre las muestras. Pusimos a prueba algunas hipótesis que podrían explicar esta diversidad. No encontramos coincidencia ni en la presencia de bacterias ni en las especies encontradas entre múltiples muestras obtenidas de un mismo macho. Tampoco encontramos correspondencia entre las bacterias presentes en las muestras de semen y de la cloaca de un mismo macho, ni entre las muestras colectadas de los machos y de las hembras que se encuentran anidando en su territorio. El estudio de muestras de eyaculaciones provenientes de tres localidades, Kentucky, New York, y Wisconsin, produjo patrones similares de prevalencia y diversidad. Muestras de semen de machos que habitan ambientes relativamente secos en tierras altas presentaron números de bacterias y especies similares a las muestras colectadas de machos que habitan territorios pantanosos. Probablemente, muchas de las bacterias son benignas, pero en ciertas circunstancias se han registrado efectos patológicos de algunas especies. Nuestros resultados sugieren que adquirir bacterias patológicas es un posible costo del apareamiento múltiple de las hembras, pero los posibles beneficios de algunos microbios podrían compensar estos riesgos.

Evaluation of antibiotic effects on phosphorus digestibility and utilization by growing-finishing pigs fed a phosphorus-deficient, corn-soybean meal diet

Evaluations of the nutritional impact of antibiotics have largely centered on effects related to the digestibility and utilization of protein and energy. Recent research has demonstrated that virginiamycin increases P digestibility. Because of the importance of P in diet cost and in waste management plans, the present study evaluated the potential impact of 2 additional antibiotics, bacitracin methylene disalicylate (bacitracin) and tylosin, on P digestibility in swine. A total of 48 barrows (mean initial BW, 63.0 to 82.9 kg) were used in 2 nutrient balance experiments. A basal corn-soybean meal diet that was not supplemented with any inorganic source of P was used in each experiment. In Exp. 1, two diets were tested: basal vs. basal plus 33.1 mg of bacitracin/kg of diet. In Exp. 2, two diets were also tested: basal vs. basal plus 44.1 mg of tylosin/kg of diet. In both experiments, the pigs were fed their diets for a minimum of 12 d before fecal and urine collection, and pigs were fed the diet at 2.7% of BW during the adaptation and collection period. In Exp. 1, the apparent DM, Ca, and P digestibility values for the basal and bacitracin diets were 91.69, 65.96, and 43.03 vs. 91.47, 65.46, and 41.79%, respectively, and did not differ by diet. In Exp. 2, the DM, Ca, and P digestibility values for the basal and tylosin diets were 91.03, 62.17, and 38.80 vs. 91.11, 63.20, and 40.10%, respectively, and did not differ by diet. The effect of the antibiotics on gut microflora was also appraised but the evaluations failed to demonstrate an effect on the microflora measured, with the exception that tylosin decreased the number of phytate-utilizing bacteria (P = 0.05). Therefore, because these 2 antibiotics did not demonstrate an improvement in P digestibility, improvements in P digestibility seem to be an antibiotic-specific response rather than a generalized antibiotic response.