Evaluation of a Preharvest Bacteriophage Therapy for Control of Salmonella within Bovine Peripheral Lymph Nodes.

ABSTRACT: A series of proof-of-concept studies were developed to determine whether a commercial bacteriophage cocktail could be utilized for the mitigation of Salmonella in bovine peripheral lymph nodes (LNs). The first objective sought to determine whether exogenous phage could be isolated from the LNs following administration. If isolation were successful, the second objective was to determine whether the phage in the LNs could effectively reduce Salmonella. Salmonella Montevideo was inoculated intradermally at multiple sites and multiple times, followed by delivery of the phage cocktail subcutaneously in two injections around each of the right and left prescapular and subiliac LNs. At the conclusion of each study, animals were euthanized, and the popliteal, prescapular, and subiliac LNs were examined. The inoculated phage was successfully isolated from the LNs; transmission electron microscopy revealed phages in the LNs of the treated cattle, and these phages were identical to those in the cocktail. Levels of phage were higher (P < 0.01) in the prescapular and subiliac LNs in the phage-treated than in the control cattle. In subsequent studies, the protocols were modified to increase Salmonella and phage levels within the LNs. Compared with the first study, overall Salmonella levels were increased in the LNs, and phage treatment decreased (P < 0.01) Salmonella in the some of the LNs. Phage levels were numerically but not significantly increased (P = 0.12) in the treated cattle. The final study was modified, hypothesizing that a 48-h postmortem period before LN removal would facilitate phage-Salmonella interaction; however, no differences (P > 0.10) in Salmonella levels were found among treatments. Salmonella-specific phages administered to live cattle can translocate to the LNs; however, these phages had limited to no effect on Salmonella in these LNs under these experimental conditions.

Robust detection of individual forensic profiles in DNA mixtures.

For a forensic identification method to be admissible in international courts, the probability of false match must be quantified. For comparison of individuals against complex mixtures using a panel of single nucleotide polymorphisms (SNPs), the probability of a random man not excluded, P(RMNE) is one admissible standard. While the P(RMNE) of SNP alleles has been previously studied, it remains to be rigorously defined and calculated for experimentally genotyped mixtures. In this report, exact P(RMNE) values were calculated for a range of complex mixtures, verified with Monte Carlo simulations, and compared alongside experimentally determined detection probabilities.

Variable response to exogenous Lactobacillus acidophilus NCFM consumed in different delivery vehicles.

AIMS: To study the effects of the delivery vehicle for Lactobacillus acidophilus on the human faecal microbiota. Our hypotheses were that (i) the delivery vehicle would influence faecal lactobacilli numbers and (ii) consumption of Lact. acidophilus would influence the populations of Bifidobacterium and hydrogen sulphide-producing bacteria. METHODS AND RESULTS: Ten subjects each received Lact. acidophilus with skim milk or water. Lactobacillus, Bifidobacterium and hydrogen sulphide-producing bacterial populations were analysed before, during and after each treatment. Regardless of the vehicle, faecal lactobacilli populations changed during treatment. Bifidobacteria and the hydrogen sulphide-producing bacteria underwent no statistically significant population changes. Intra- and intersubject variability was observed. CONCLUSIONS: The vehicle in which Lact. acidophilus was delivered did not influence faecal lactobacilli numbers. Consumption of Lact. acidophilus did not influence the populations of Bifidobacterium and hydrogen sulphide-producing bacteria. The lactobacilli populations of subjects were variable. The fed lactobacilli did not appear to colonize the gastrointestinal tract. SIGNIFICANCE AND IMPACT OF THE STUDY: We provide evidence that (i) there was no collective advantage to using skim milk as a delivery vehicle vs water; (ii) exogenous Lact. acidophilus did not affect endogenous bifidobacteria or hydrogen sulphide-producing bacteria; (iii) data should be carefully examined before pooling for analysis and (iv) continuous feeding was required to maintain an elevated lactobacilli population.

Genome of the bacterium Streptococcus pneumoniae strain R6.

Streptococcus pneumoniae is among the most significant causes of bacterial disease in humans. Here we report the 2,038,615-bp genomic sequence of the gram-positive bacterium S. pneumoniae R6. Because the R6 strain is avirulent and, more importantly, because it is readily transformed with DNA from homologous species and many heterologous species, it is the principal platform for investigation of the biology of this important pathogen. It is also used as a primary vehicle for genomics-based development of antibiotics for gram-positive bacteria. In our analysis of the genome, we identified a large number of new uncharacterized genes predicted to encode proteins that either reside on the surface of the cell or are secreted. Among those proteins there may be new targets for vaccine and antibiotic development.

Sublethal sanitizer stress and adaptive response of Escherichia coli O157:H7.

The effect of sublethal exposure to peroxyacetic acid (PAA) sanitizer on adaptation to peroxidative stress and development of thermal cross-resistance was investigated in Escherichia coli O157:H7. Acute sublethal PAA sanitizer exposure was used to represent a contact scenario. Cultures were grown in Trypticase soy-yeast extract broth. Acute treatment cultures were pretreated with 0.1% PAA, then all cultures were challenged at either 80 mM H202 or 54 degrees C. Acute and peroxide control cultures showed substantially increased peroxidative tolerance (D80mM > 2 h) versus negative control cultures not exposed to sanitizer (D80mM = 0.19+/-0.03 h). The inactivation rate of the acetic acid control (D80mM = 0.21+/-0.05 h) was similar to the negative control rate. Acute (D54 degrees C = 0.55+/-0.07 h) cultures did not exhibit increased thermal resistance versus the control (D54 degrees C = 0.54+/-0.07 h). Thermal injury was determined as difference in D54 degrees C value (deltaD54 degrees c) obtained on pyruvate and deoxycholate media. Thermal-induced injury was not observed in either control (deltaD54 degrees C = 0.04 h) or acute (deltaD54 degrees C = 0.05 h) cultures.

Physician shortage areas and policies to influence practice location.

Our intention is to examine the common assumption that physician supply is related strongly to more direct measures of economic shortage. The concept of a medical shortage defined by untreated illness or excess rates of preventable illness is not considered here. We examine the more policy-specific issue of whether the present shortage areas include the most extreme areas, based on these indicators of market tightness. If supply is a poor predictor of shortage, then more direct measures or other more carefully targeted policies should be explored.

High-cost users of medical care.

Cost characteristics of hospital patients were analyzed in 2238 medical records randomly selected from 42,880 dicharges in six contrasting hospital populations in the year 1976. Total hospital billings were concentrated on a few patients. On average, the high-cost 13% of patients consumed as many resources as the low-cost 87%. Repeated hospitalizations for the same disease were more characteristic of the expensive patients than were single cost-intensive stays, "intensive care," or prolonged single hospitalizations. Potentially harmful personal habits (e.g., drinking and smoking) were indicated in the records of high-cost patients substantially more often than in those of low-cost patients. Unexpected complications during treatment were five times more frequent in the high-cost group. Public policy programs for health insurance or cost control should include provisions based on the special characteristics of high-cost patients.

Repeated hospitalization for the same disease: a multiplier of national health costs.

In five typical hospitals, and a specialized treatment center, more than half of all patients--and 60 percent of all costs--were attributable to repeated admissions for the same disease. The fiscal and clinical nature of repeated hospitalization suggests changes for national health policy. These will require more sensitive identification of patients "at risk" of recidivism, and insurance mechanisms that relate more rationally and equitably to long-term needs.