PubMed Health⌕ Search

Biomedical subjects

M P Doyle

Publications and source records attributed to M P Doyle.

At least 55 records · Page 3Linked to original sources

Inactivation of Escherichia coli O157:H7 and Listeria monocytogenes on plastic kitchen cutting boards by electrolyzed oxidizing water.

One milliliter of culture containing a five-strain mixture of Escherichia coli O157:H7 (approximately 10(10) CFU) was inoculated on a 100-cm2 area marked on unscarred cutting boards. Following inoculation, the boards were air-dried under a laminar flow hood for 1 h, immersed in 2 liters of electrolyzed oxidizing water or sterile deionized water at 23 degrees C or 35 degrees C for 10 or 20 min; 45 degrees C for 5 or 10 min; or 55 degrees C for 5 min. After each temperature-time combination, the surviving population of the pathogen on cutting boards and in soaking water was determined. Soaking of inoculated cutting boards in electrolyzed oxidizing water reduced E. coli O157:H7 populations by > or = 5.0 log CFU/100 cm2 on cutting boards. However, immersion of cutting boards in deionized water decreased the pathogen count only by 1.0 to 1.5 log CFU/100 cm2. Treatment of cutting boards inoculated with Listeria monocytogenes in electrolyzed oxidizing water at selected temperature-time combinations (23 degrees C for 20 min, 35 degrees C for 10 min, and 45 degrees C for 10 min) substantially reduced the populations of L. monocytogenes in comparison to the counts recovered from the boards immersed in deionized water. E. coli O157:H7 and L. monocytogenes were not detected in electrolyzed oxidizing water after soaking treatment, whereas the pathogens survived in the deionized water used for soaking the cutting boards. This study revealed that immersion of kitchen cutting boards in electrolyzed oxidizing water could be used as an effective method for inactivating foodborne pathogens on smooth, plastic cutting boards.

Colony Count, Microbial↗

Control of enantioselectivity in catalytic metal carbene reactions.

Chiral dirhodium(II) complexes constructed from 2-oxopyrrolidine, 2-oxazolidinone, N-acyl-2-imidazolidinone, or 2-azetidinone ligands are exceptional catalysts for enantioselective metal carbene transformations which provide lactones and lactams via cyclopropanation, cyclopropenation, C-H insertion, and ylide reactions with enantiomeric excesses greater than 90% and, usually, in very high yields. These chiral catalysts present high diastereo-, regio- and chemoselectivities, turnover numbers up to 1000, as well as recoverability and reuse.

Catalysis↗

Virulence genes of Shiga toxin-producing Escherichia coli isolated from food, animals and humans.

The presence of virulence genes, encoding enterohemorrhagic Escherichia coli (EHEC)-hemolysin (EHEC-hlyA), intimin (eae), and Shiga toxins 1 (stx1) and 2 (stx2), in 178 isolates of pathogenic E. coli, was determined using the polymerase chain reaction with primers specific for each virulence gene. The tested organisms were 120 isolates of E. coli O157:H7 from human patients, cattle, sheep and foods, 16 non-O157:H7 EHEC isolates from patients suffering from hemorrhagic colitis or hemolytic uremic syndrome, 15 non-O157:H7 Shiga toxin-producing E. coli (STEC) isolates from cattle and foods, 26 isolates of enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC) and enterotoxigenic E. coli (ETEC), and an E. coli K12 strain. Results revealed that all isolates of O157:H7 carried EHEC-hlyA, eae, and one or both stx genes; 15 of the 16 non-O157:H7 EHEC isolates had EHEC-hlyA, but all possessed eae and one or both stx genes; only seven of the 15 non-O157 STEC isolated from cattle and foods contained both EHEC-hlyA and eae genes. The EPEC, EIEC, ETEC, and the E. coli K12 strain did not carry these virulence genes, except eight EPEC isolates were positive for eae. Results suggest that a combination of EHEC-hlyA and eae genes could serve as markers to differentiate EHEC from less pathogenic STEC, and other pathogenic or non-pathogenic E. coli.

Adhesins, Bacterial↗

Rate of reaction with nitric oxide determines the hypertensive effect of cell-free hemoglobin.

Administration of extracellular hemoglobin-based oxygen carriers often induces mild increases in blood pressure. In order to test whether nitric oxide (NO) scavenging is responsible for the hypertensive effect, we constructed and tested a set of recombinant hemoglobins that vary in rates of reaction with NO. The results suggest that the rapid reactions of oxy- and deoxyhemoglobin with nitric oxide are the fundamental cause of the hypertension. The magnitude of the blood-pressure effect correlates directly with the in vitro rate of NO oxidation. Hemoglobins with decreased NO-scavenging activity may be more suitable for certain therapeutic applications than those that cause depletion of nitric oxide.

Animals↗

Heat shock response enhances acid tolerance of Escherichia coli O157:H7.

Escherichia coli O157:H7 (O157) has unusual acid tolerance. The influence of heat shock on acid tolerance of O157 was studied. Seven strains of O157 and E. coli K-12 were tested for their ability to survive in minimum glucose medium (pH 2.5) at 37 degrees C. The survival of heat-shocked (10 min at 48 degrees C) cells was about 10-100 times greater compared with untreated cells depending on the strain. No significant difference (P > 0.05) for O157 strain 932 was observed between heat shock-induced and acid adaptation-induced (pH 5.0) acid tolerance. Chloramphenicol prevented heat shock-induced acid tolerance, indicating the requirement of newly synthesized protein(s). Two outer membrane proteins (OMP) (22 and 15 kDa) were synthesized within 10 min of heat shock and were expressed for at least 6 h by cells held at 37 degrees C. N-terminal amino acid sequence analysis suggested that the 22 kDa OMP is a component of an alkyl hydroperoxide reductase. This protein contains a redox active disulphide, which is probably involved in H+ transport. Results indicate that sublethal heat treatment of O157 cells substantially increases their tolerance to acidic conditions. This could have practical implications for foods that receive a mild heat treatment and rely on acid as a preservative.

Amino Acid Sequence↗

Reduction of carriage of enterohemorrhagic Escherichia coli O157:H7 in cattle by inoculation with probiotic bacteria.

Bacteria inhibitory to Escherichia coli O157:H7 were isolated from cattle and evaluated for their potential for reducing carriage of E. coli O157:H7 in calves. Eighteen of 1,200 bacterial isolates from cattle feces and intestinal tissue samples were screened and determined to inhibit the growth of E. coli O157:H7 in vitro. Seventeen of the isolates were E. coli and one was Proteus mirabilis. None produced Shiga toxin. Genomic DNA fingerprinting by pulsed-field gel electrophoresis revealed 13 distinguishable profiles among the 18 isolates. Two calves inoculated perorally with a mixture of all 18 isolates (10(10) CFU) appeared to be normal and did not develop signs of clinical disease throughout a 25- to 27-day observation period. These bacteria colonized segments of the gastrointestinal tract and were in feces at the termination of the experiment (25 and 27 days postinoculation) at levels of 50 to 200 CFU/g. Fifteen cannulated calves were studied to determine the efficiency of the probiotic bacteria in reducing or eliminating the carriage of E. coli O157:H7. Nine calves served as controls, with each animal receiving perorally 10(10) CFU of E. coli O157:H7. E. coli O157:H7 was detected intermittently in the rumen samples from all control animals throughout 3 weeks postinoculation, whereas E. coli O157:H7 was shed at various levels in feces continuously throughout the experiment (mean, 28 days). E. coli O157:H7 was isolated from the rumens and colons of eight of nine and nine of nine calves, respectively, at the termination of the study. Six calves each received perorally 10(10) CFU of probiotic bacteria and then 2 days later received 10(10) CFU of E. coli O157:H7. E. coli O157:H7 was detected in the rumen for only 9 days postinoculation in two animals, for 16 days in one animal, for 17 days in two animals, and for 29 days in one animal. E. coli O157:H7 was detected in feces for only 11 days postinoculation in one animal, for 15 days in one animal, for 17 days in one animal, for 18 days in one animal, for 19 days in one animal, and for 29 days in one animal. At the end of the experiment (mean, 30 days), E. coli O157:H7 was not recovered from the rumen of any of the six animals treated with probiotic bacteria; however, E. coli O157:H7 was recovered from the feces of one of the animals. This animal was fasted twice postinoculation. These studies indicate that selected probiotic bacteria administered to cattle prior to exposure to E. coli O157:H7 can reduce the level of carriage of E. coli O157:H7 in most animals.

Animals↗

Preclinical biology of recombinant human hemoglobin, rHb1.1.

Historically, the development of hemoglobin based oxygen carriers, HBOCs, were confounded by issues related to activation of the complement cascade and other inflammatory processes, renal toxicity, and significant systemic vasoconstriction. However, with shortages in the blood supply, the risk of infectious agent contamination, and delays associated with complete crossmatch as well as transfusion reactions, HBOC development has assumed greater importance. A successful HBOC in addition to having favorable oxygen binding parameters and colloid oncotic properties, must also have a low toxicity profile, be nonimmunogenic, have positive rheologic properties, and have an adequate in vivo half life. In addition, it must also be stable in vivo and not undergo significant oxidation to methemoglobin or release heme or iron in the vasculature. The preclinical studies which have been designed and executed to address these requirements for recombinant human hemoglobin rHb1.1 serve as the focus of this review. Recombinant Hb1.1 represents the first HBOC to enter clinical trials as a recombinant product in distinction to other HBOCs which are derived from bovine or outdated human blood. While currently in phase II clinical trials, the preclinical biology which has increased our understanding of this molecule are the subject of this review.

Animals↗

Antibiotic resistance of Escherichia coli O157:H7 and O157:NM isolated from animals, food, and humans.

Antibiotic resistance was determined for 118 E. coli O157:H7 and 7 O157:NM isolates from animals, foods, and humans. Among the 125 isolates, 30 (24%) were resistant to at least one antibiotic and 24 (19%) were resistant to three or more antibiotics. Cattle isolates had the highest rate (34%) of antibiotic resistance. The seven resistant food isolates were all from ground beef. The most frequent resistance type overall was streptomycin-sulfisoxazole-tetracycline, which accounted for over 70% of the resistant strains. Two E. coli O157:NM isolates from cattle were resistant to six antibiotics: ampicillin, kanamycin, sulfisoxazole, streptomycin, tetracycline, and ticarcillin. Streptomycin was the most common antibiotic to which E. coli O157:H7 and O157:NM were resistant (29 out of 30 isolates), followed by tetracycline (26 isolates). This study suggests that E. coli O157:H7 and O157:NM have developed resistance to antibiotics. Research is needed to define mechanisms of antibiotic resistance in E. coli O157:H7 and to minimize the development of resistance.

Animals↗

Thermal inactivation of Escherichia coli O157:H7 isolated from ground beef and bovine feces, and suitability of media for enumeration.

Rates of thermal inactivation of five strains of Escherichia coli O157:H7 isolated from ground beef implicated in outbreaks of hemorrhagic colitis and five strains isolated from bovine feces were determined. Ground beef (22% fat, 10 g), inoculated with individual test strains at populations ranging from 6.85 to 7.40 log10 CFU g-1 of beef, was formed into patties (0.3 cm thick and 8.0 cm in diameter) and sealed in polyethylene bags. For each strain and treatment temperature (54.4, 58.9, 62.8, 65.6, or 68.3 degrees C), 6 bags were simultaneously immersed into a recirculating water bath. Viable cells in patties heated for various lengths of time were enumerated by plating diluted samples on sorbitol MacConkey agar supplemented with 4-methylumbelliferyl-beta-D-glucuronide (MSMA) and modified eosin methylene blue (MEMB) agar. Regardless of strain or treatment temperature, higher numbers of E. coli O157:H7 cells were generally recovered on MEMB agar than on MSMA, indicating the inferiority of MSMA as a recovery medium for quantitative determination of E. coli O157:H7 cells in heat-processed ground beef. Significantly (P < or = 0.05) higher D values when enumeration was done using MEMB agar compared with MSMA. Mean D values for combined strain data at 54.4, 58.9, 62.8, and 65.6 degrees C from cultures on MEMB agar were 123.90, 6.47, 0.62, and 0.20 min, respectively, whereas D values of 25.5, 5.21, and 0.18 min were obtained at the same temperatures from cultures on MSMA. Results suggest that cooking ground beef patties to an internal temperature of 68.3 degrees C for 40 s will inactivate at least 99.99% of E. coli O157:H7 cells; z values of 4.0 and 5.1 degrees C were calculated from mean D values obtained from MEMB agar and MSMA, respectively, as recovery media. Differences in D values existed among strains but rates of thermal inactivation do not appear to be correlated with the sources of the isolates.

Animals↗

Inhibition of Listeria monocytogenes and Aeromonas hydrophila by plant extracts in refrigerated cooked beef.

Refrigerated ready-to-eat foods are becoming increasingly popular but are often vulnerable to contamination and subsequent growth by psychrotrophic foodborne pathogens. Consequently, there is a need for additional methods to assure the safety of these foods. Beef slices prepared from roasted whole sirloin tips were used in the study. Nine plant extracts were evaluated for ability to inhibit the growth of two psychrotrophic pathogens (Aeromonas hydrophila and Listeria monocytogenes) in refrigerated cooked beef. Results indicated that only eugenol (clove extract) and pimento extract significantly inhibited the growth of A. hydrophila and L. monocytogenes. However, L. monocytogenes was not as sensitive as was A. hydrophila to both treatments, especially to pimento extracts. These results suggest that plant extracts might be useful as an antimicrobial in cooked ready-to-eat meat.

Aeromonas hydrophila↗

Survival of enterohemorrhagic Escherichia coli O157:H7 in water.

Several recent Escherichia coli O157:H7 outbreaks associated with both drinking and recreational water raise concerns about waterborne illness caused by this pathogen. The survival characteristics of a mixture of five nalidixic acid-resistant E. coli O157:H7 strains (10(3) CFU/ml) in filtered and autoclaved municipal water, in reservoir water, and in water from two recreational lakes were determined for a period of 91 days at 8, 15 or 25 degrees C. Greatest survival was in filtered autoclaved municipal water and least in lake water. Regardless of the water source, survival was greatest at 8 degrees C and least at 25 degrees C. E. coli O157:H7 populations decreased by 1 to 2 log10 by 91 days at 8 degrees C, whereas the pathogen was not detectable (> or 3 = log10 decrease) within 49 to 84 days at 25 degrees C in three of the four water sources. SDS-PAGE of surface antigens of surviving cells revealed that there was no major alteration in lipopolysaccharide pattern, but outer membrane protein composition did change. These studies indicate that E. coli O157:H7 is a hardy pathogen that can survive for long periods of time in water, especially at cold temperatures. However, direct viable counts of E. coli O157:H7 determined by acridine orange staining remained essentially the same for 12 weeks at 25 degrees C, whereas viable counts on tryptic soy agar plates decreased to undetectable levels within 12 weeks. Results suggest that E. coli O157:H7 can enter a viable but nonculturable (VBNC) state in water.

Antigens, Bacterial↗

Effect of environmental and substrate factors on survival and growth of Helicobacter pylori.

The effect of temperature (4 to 42 degrees C), NaCl concentration (0.5 to 7.5%), NaNO2 concentration (0 to 400 micrograms/ml), water activity (aw level of 0.6 to 0.995), pH (3.5 to 7.3) and urea (8 mM) on the survival and growth of Helicobacter pylori in a nutrient-rich laboratory culture medium was investigated. Under microaerobic conditions (5% O2, 10% CO2, and 85% N2), the organism grew well in brain heart infusion broth supplemented with 7% horse serum and antibiotics (BHI-HS-TVA) in a temperature range of 30 to 37 degrees C with agitation. H. pylori (initial population of ca. 5 x 10(3) CFU/ml) survived for 14 days at 4 degrees C, for 2 days at 25 degrees C, and for less than 1 day at 40 and 42 degrees C. The optimal NaCl concentration for growth of H. pylori was 0.5 to 1.0%; 2.0% NaCl inhibited growth. Up to 400 micrograms of NaNO2 per ml did not prevent growth. The minimum aw (adjusted with glycerol) and pH (acidified with HCl) for growth of H. pylori was 0.98 and 4.5, respectively. The addition of urea to broth greatly enhanced the growth of H. pylori at both pH 4.5 and 5.5. Although H. pylori did not grow at pH 3.5, the presence of urea in broth enhanced its survival. Considering the apparent fastidious conditions for growth of H. pylori in BHI-HS-TVA broth, H. pylori is unlikely to grow well, if at all, in most foods. The bacterium may, however, survive for extended periods of time in low acid-high moisture environments under refrigerated storage.

Helicobacter pylori↗

Development of a model for evaluation of microbial cross-contamination in the kitchen.

Foods can become contaminated with pathogenic microorganisms from hands, the cutting board, and knives during preparation in the kitchen. A laboratory model was developed to determine occurrence of cross-contamination and efficacy of decontamination procedures in kitchen food-handling practices. Enterobacter aerogenes B199A, an indicator bacterium with attachment characteristics similar to that of Salmonella spp., was used. Chicken meat with skin inoculated with 10(6) CFU of E. aerogenes B199A/g was cut into small pieces on a sterile cutting board. The extent of cross-contamination occurring from meat to the cutting board and from the cutting board to vegetables (lettuce and cucumbers) subsequently cut on the board was determined. Swab samples from the cutting board, hand washings, and lettuce and cucumber samples revealed that approximately 10(5) CFU of E. aerogenes/cm2 were transferred to the board and hands and approximately 10(3) to 10(4) CFU of E. aerogenes/g to the lettuce and cucumbers. The surfaces of the cutting board and hands were treated with antibacterial agents after cutting the meat, and counts of E. aerogenes on the cutting board and vegetables (lettuce and cucumbers) were determined. Results revealed that use of the disinfectant reduced the population of E. aerogenes to almost nondetectable levels on the cutting boards. The average counts after treatment were < 20 CFU/g of vegetable and ranged from < 20 to 200 CFU per cm2 or g on the cutting board and subsequently on the vegetables. These results indicate that bacteria with attachment characteristics similar to Salmonella spp. can be readily transferred to cutting boards during food preparation and then cross-contaminate fresh vegetables if the boards are not cleaned. Application of a kitchen disinfectant can greatly reduce bacterial contamination on cutting boards.

Bacterial Adhesion↗

Microbiological quality and the inability of proteolytic Clostridium botulinum to produce toxin in film-packaged fresh-cut cabbage and lettuce.

The production of toxin by a 10-strain mixture of proteolytic Clostridium botulinum in fresh produce packaged in polyethylene films having high (7,000 cc/m2/24 h; HOTR) and low (3,000 cc/m2/24 h; LOTR) relative oxygen permeability was determined. Shredded cabbage and lettuce inoculated with approximately 10(2) spores/g were placed in bags composed of the two films (1.4 kg/bag), and the bags were then vacuum sealed. Produce was stored at 4, 13, and 21 degrees C for up to 21 (cabbage) or 28 (lettuce) days and analyzed periodically. At each sampling time, the gas composition within the bags, pH of the produce, and microbial populations (total aerobic and anaerobic microorganisms, lactic acid bacteria, psychrotrophic bacteria, and yeasts and molds) were determined. In addition, the presence of botulinal toxin was determined using the standard U.S. Food and Drug Administration mouse bioassay protocol. Bags made of HOTR film prolonged sensory quality of cabbage and lettuce, especially at 13 and 4 degrees C. Packaging material had an effect on the growth of various groups of microorganisms; however, there was not a general trend. For example, lettuce packaged in HOTR bags had higher aerobic microbial populations than that packed in LOTR, but no significant difference (P < or = 0.05) was observed with cabbage. Growth of psychrotrophic bacteria was greater in vegetables packaged in HOTR film while growth of yeasts and molds was not affected by either packaging film. Most differences in microbial populations in produce packaged in LOTR and HOTR films were less than 1 log10 CFU/g. Botulinal toxin was not detected in cabbage or lettuce packaged in either film or stored under any test condition.

Animals↗

Elevation of intracellular calcium in smooth muscle causes endothelial cell generation of NO in arterioles.

It is well known that vascular smooth muscle tone can be modulated by signals arising in the endothelium (e.g., endothelium-derived relaxing factor, endothelium-derived hyperpolarizing factor, and prostaglandins). Here we show that during vasoconstriction a signal can originate in smooth muscle cells and act on the endothelium to cause synthesis of endothelium-derived relaxing factor. We studied responses to two vasoconstrictors (phenylephrine and KCl) that act by initiating a rise in smooth muscle cell intracellular Ca2+ concentration ([Ca2+]i) and exert little or no direct effect on the endothelium. Fluo-3 was used as a Ca2+ indicator in either smooth muscle or endothelial cells of arterioles from the hamster cheek pouch. Phenylephrine and KCl caused the expected rise in smooth muscle cell [Ca2+]i that was accompanied by an elevation in endothelial cell [Ca2+]i. The rise in endothelial cell [Ca2+]i was followed by increased synthesis of NO, as evidenced by an enhancement of the vasoconstriction induced by both agents after blockade of NO synthesis. The molecule involved in signal transmission from smooth muscle to endothelium is as yet unknown. However, given that myoendothelial cell junctions are frequent in these vessels, we hypothesize that the rise in smooth muscle cell Ca2+ generates a diffusion gradient that drives Ca2+ through myoendothelial cell junctions and into the endothelial cells, thereby initiating the synthesis of NO.

Acetylcholine↗

An outbreak of Escherichia coli O157:H7 infections traced to jerky made from deer meat.

OBJECTIVE: To investigate a 1995 outbreak of Escherichia coli O157:H7 infections and to assess the safety of meat dehydration methods. DESIGN: Survey subsequent to routine surveillance report, environmental investigations, and laboratory experimentation. SETTING: Oregon community. PARTICIPANTS: Members of an extended household and their social contacts with confirmed or presumptive E coli O157:H7 infections. RESULTS: A total of 6 confirmed and 5 presumptive cases were identified. Homemade venison jerky was implicated as the source of transmission. E coli O157:H7 with the same distinctive, pulsed-field gel electrophoresis pattern seen in the case isolates was recovered from leftover jerky, uncooked meat from the same deer, a saw used to dismember the carcass, and fragments of the deer hide. In a subsequent survey, E coli O157:H7 was recovered from 3 (9%) of 32 deer fecal pellets collected in nearby forest land. In the laboratory, inoculated venison was dried at several time and temperature combinations, ranging up to 10 hours at 62.8 degrees C. Viable organisms were recovered under all conditions tested. CONCLUSIONS: Deer can be colonized by E coli O157:H7 and can be a source of human infections. Conditions necessary to ensure the safety of dried meat deserve further review. Game should be handled with the same caution indicated for commercially slaughtered meat.

Adolescent↗

A multiplex PCR for identifying Shiga-like toxin-producing Escherichia coli O157:H7.

A multiplex PCR assay specifically detecting Escherichia coli O157:H7 was developed by employing primers amplifying a DNA sequence upstream of E. coli O157:H7 eaeA gene and genes encoding Shiga-like toxins (SLT) I and II. Analysis of 151 bacterial strains revealed that all E. coli O157:H7 strains were identified simultaneously with the SLT types and could be distinguished from E. coli O55:H7 and E. coli O55:NM, and other non-O157 SLT-producing E. coli strains. Primer design, reaction composition (in particular, primer quantity and ratios), and amplification profile were most important in development of this multiplex PCR. This assay can serve not only as a confirmation test but also potentially can be applied to detect the pathogen in food.

Adhesins, Bacterial↗

Experimental Escherichia coli O157:H7 carriage in calves.

Nine weaned calves (6 to 8 weeks of age) were given 10(10) CFU of a five-strain mixture of enterohemorrhagic Escherichia coli O157:H7 by oral-gastric intubation. After an initial brief period of pyrexia in three calves and transient mild diarrhea in five calves, calves were clinically normal throughout the 13- to 27-day study. The population of E. coli O157:H7 in the faces decreased dramatically in all calves during the first 2 weeks after inoculation. Thereafter, small populations of E. coli O157:H7 persisted in all calves, where they were detected intermittently in the feces and rumen contents. While withholding food increased fecal shedding of E. coli O157:H7 by 1 to 2 log10/g in three of four calves previously shedding small populations of E. coli O157:H7, the effect of fasting on fecal shedding of E. coli O157:H7 was variable in calves shedding larger populations. At necropsy, E. coli O157:H7 was not isolated from sites outside the alimentary tract. E. coli O157:H7 was isolated from the forestomach or colon of all calves at necropsy. Greater numbers of E. coli O157:H7 were present in the gastrointestinal contents than in the corresponding mucosal sections, and there was no histologic or immunohistochemical evidence of E. coli O157:H7 adhering to the mucosa. In conclusion, under these experimental conditions, E. coli O157:H7 is not pathogenic in weaned calves, and while it does not appear to colonize mucosal surfaces for extended periods, E. coli O157:H7 persists in the contents of the rumen and colon as a source for fecal shedding.

Animals↗