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Biomedical subjects

J M Sargeant

Publications and source records attributed to J M Sargeant.

At least 19 recordsLinked to original sources

The process of systematic review and its application in agri-food public-health.

Foodborne illness is an important public-health concern. Livestock operations represent the original source of many foodborne pathogens. Policy- and decision-makers in the area of agri-food public-health need dependable information on interventions to reduce foodborne diseases. While individual studies might be weak or reach contradictory conclusions, the body of information contained in all studies might reveal a clearer picture of the state of knowledge. A formal process has been developed for conducting systematic reviews of the human healthcare literature. Systematic reviews differ from traditional narrative reviews in that systematic reviews follow a structured research protocol to reduce sources of bias at all stages of the review. This includes the objective, the literature search, the identification of relevant literature, quality assessment of relevant studies, summarization or statistical analysis of data, and conclusions. To date, systematic reviews have not been widely used in agri-food public-health. Existing systematic reviews on agri-food public-health topics revealed a scarcity of rigorously conducted intervention research, indicating a need for additional high quality primary studies. The specific protocols used to conduct systematic reviews in agri-food public-health need to accommodate the use of challenge studies and observational studies, the lack of randomized controlled trials, and the clustering of animal populations.

Animals↗

Pulsed-field gel electrophoresis patterns of Escherichia coli O157 isolates from Kansas feedlots.

This study investigated the prevalence and distribution of Escherichia coli O157 genetic types within and among feedlots using pulsed-field gel electrophoresis to separate XbaI-digested DNA. The study population consisted of 300 pens of cattle in 30 feedlots in Kansas that were sampled (feces, water, and water sediment) within a month of being shipped for slaughter. The prevalence of E. coli O157 was 8.5% in feces, 3.1% in water, and 4.5% in water sediment samples. A total of 424 E. coli O157 isolates were characterized by pulsed-field gel electrophoresis, and 139 subtypes (100% Dice similarity with no band differences) were identified. The majority of subtypes (70/139) was identified only once, but nine were identified 10 or more times. Identical subtypes were recovered from both feces and water tanks in 10 feedlots. The majority of subtypes were identified in only one feedlot, and the number of subtypes ranged from one to 23 within a feedlot and from one to seven within a pen. There were 10 feedlots with at least 15 positive samples. In these 10 feedlots, the most common subtype accounted for 16.9-78.6% of the isolates. Common subtypes differed among feedlots. In eight of the 10 feedlots, the most common subtype was identified in multiple pens. The results support a complex ecology for E. coli O157 in feedlot operations, with factors associated with exposure and transmission likely acting at a common level for multiple feedlots, within feedlots, and within pens of cattle.

Animals↗

Effect of pooling bovine fecal samples on the sensitivity of detection of E. coli O157:H7.

To assess the effect of pooling fecal samples on the sensitivity of detection of E. coli O157:H7, 12 calves, inoculated orally with 10(8)cfu per calf of nalidixic acid resistant E. coli O157:H7, were used to provide positive fecal samples. After inoculation, calves were sampled twice weekly. Negative fecal samples were from calves at a local dairy. Samples from inoculated calves were incubated without pooling or were mixed with known negative fecal samples in a 1:4 ratio or a 2:3 ratio (positive:negative) for detection of E. coli O157:H7. Samples were enriched 6h in Gram negative broth with vancomycin, cefixime, and cefsoludin, underwent immunomagnetic separation with Dynabeads, and were plated onto sorbitol MacConkey agar with cefixime, and tellurite (SMACct). Morphologically typical colonies were plated onto blood agar, incubated overnight at 37 degrees C and an indole test was performed on each colony. Indole positives colonies were plated on SMAC agar with 20 microg/ml nalidixic acid (SMACnal). Colonies that grew on SMACnal were confirmed by O157 agglutination. Sensitivity of detection in non-pooled samples was 77%. Samples pooled 1:4 and 2:3 with negative samples were 55 and 52% sensitive, respectively. Pooling decreased sensitivity of detection for E. coli O157:H7 in bovine fecal samples (P<0.01). A deterministic binomial probability model was developed to assess the probability of detecting pens of cattle shedding E. coli O157 using a pooling protocol or individual samples. Pooling decreased sensitivity of detection at low pen prevalence compared to individual samples but was similar at high prevalence.

Animals↗

Comparison of rectoanal mucosal swab cultures and fecal cultures for determining prevalence of Escherichia coli O157:H7 in feedlot cattle.

We compared fecal samples with samples collected with rectoanal mucosa swabs (RAMS) to determine the prevalence of Escherichia coli O157 in feedlot cattle (n = 747). Escherichia coli O157 was detected in 9.5% of samples collected with RAMS and 4.7% of samples tested by fecal culture. Pulsed-field gel electrophoresis analysis of isolates suggested that the strains colonizing the rectoanal junction were the same as those from the feces. Mucosal swab sampling was more sensitive than fecal sampling for determining the prevalence of E. coli O157 in feedlot cattle.

Anal Canal↗

Association of cow and quarter-level factors at drying-off with new intramammary infections during the dry period.

Our objective was to describe cow and quarter-level factors associated with drying-off, and to evaluate their impacts on new intramammary infections (IMI) during the dry period. Data from 300 cows in five research herds were collected starting 2 weeks prior to scheduled drying-off. Variables of interest included daily milk production, teat-end integrity, formation of the teat-canal keratin plug, and quarter-milk bacteriological culture results. Overall, 11% of quarters developed new IMI in the dry period; this varied by herd, parity and time of the study. Most new IMI were caused by environmental streptococci and coliform organisms (34 and 30%, respectively). Quarters that had a cracked teat-end had higher odds of developing new infections than those without cracks (15 and 10%, respectively). Quarters that formed a keratin plug early in the dry period had a lower odds than those that did not close (10 and 14%, respectively). After 6 dry weeks, 23% of quarters were still open. The hazard of quarters closing if milk production on the day prior to drying-off was >21 kg 1.8-times less.

Animals↗

Effect of forage or grain diets with or without monensin on ruminal persistence and fecal Escherichia coli O157:H7 in cattle.

Twelve ruminally cannulated cattle, adapted to forage or grain diet with or without monensin, were used to investigate the effects of diet and monensin on concentration and duration of ruminal persistence and fecal shedding of E. coli O157:H7. Cattle were ruminally inoculated with a strain of E. coli O157:H7 (10(10) CFU/animal) made resistant to nalidixic acid (Nal(r)). Ruminal and fecal samples were collected for 11 weeks, and then cattle were euthanized and necropsied and digesta from different gut locations were collected. Samples were cultured for detection and enumeration of Nal(r) E. coli O157:H7. Cattle fed forage diets were culture positive for E. coli O157:H7 in the feces for longer duration (P < 0.05) than cattle fed a grain diet. In forage-fed cattle, the duration they remained culture positive for E. coli O157:H7 was shorter (P < 0.05) when the diet included monensin. Generally, ruminal persistence of Nal(r) E. coli O157:H7 was not affected by diet or monensin. At necropsy, E. coli O157:H7 was detected in cecal and colonic digesta but not from the rumen. Our study showed that cattle fed a forage diet were culture positive longer and with higher numbers than cattle on a grain diet. Monensin supplementation decreased the duration of shedding with forage diet, and the cecum and colon were culture positive for E. coli O157:H7 more often than the rumen of cattle.

Animal Feed↗

Effect of antibiotics in milk replacer on fecal shedding of Escherichia coli O157:H7 in calves.

The objective of this study was to compare the concentration and duration of fecal shedding of Escherichia coli O157:H7 between calves fed milk replacer with or without antibiotic (oxytetracycline and neomycin) supplementation. Eighteen 1-wk-old Holstein calves were orally inoculated with a strain of E. coli O157:H7 (3.6 x 10(8) cfu/calf) made resistant to nalidixic acid (NA). Rectal samples were obtained three times weekly for 8 wk following oral inoculation. Fecal shedding of NA-resistant E. coli O157:H7 was quantified by direct plating or detected by selective enrichment procedure. Eight weeks after inoculation, calves were killed, necropsied, and tissues (tonsils, retropharyngeal and mesenteric lymph nodes, and Peyer's patches) and gut contents (rumen, omasum, abomasum, ileum, cecum, colon, and rectum) were sampled to quantify or detect NA-resistant E. coli O157:H7. The percentage of calves shedding NA-resistant E. coli O157:H7 in the feces in the antibiotic-fed group was higher (P < 0.001) early in the study period (d 6 and 10) compared with the control group fed no antibiotics. There was no difference between treatment and control groups in the concentration of E. coli O157 in feces that were positive at quantifiable concentrations. A comparison of the duration of fecal shedding between treated and untreated calves showed no significant difference between groups. At necropsy, E. coli O157:H7 was recovered from the rumen and omasum of one calf in the control group and from retropharyngeal lymph node and Peyer's patch of two calves in the antibiotic group. Supplementation of milk replacer with antibiotics may increase the probability of E. coli O157:H7 shedding in dairy calves, but the effect seems to be of low magnitude and short duration.

Abomasum↗

Escherichia coli O157 in feedlot cattle feces and water in four major feeder-cattle states in the USA.

The prevalence of Escherichia coli O157 was determined in 10662 fecal samples, 2130 water and 1132 water tank-sediment samples collected during the summer months in 2001 from 711 pens in 73 feedlots located in Kansas, Nebraska, Texas, or Oklahoma, USA. Overall, 10.2% of fecal samples were positive for E. coli O157, with 52% of the pens and 95.9% of the feedlots having at least one positive fecal sample. There were no differences among states or months in the fecal prevalences. Water or water tank-sediment was positive in 13.1% of the water tanks, and 60.3% of feedlots had at least one positive tank. Cattle were more likely to be shedding E. coli O157 in pens with positive water tanks, and water was more likely to be positive when E. coli O157 was detected in the sediment.

Animals↗

Comparison of cultivation and PCR-hybridization for detection of Salmonella in porcine fecal and water samples.

A total of 150 fecal and water samples from four swine farms were tested for the presence of Salmonella enterica using different enrichment techniques as follows: (i) 92 fecal samples from nursery and farrowing barns at three swine farms were preenriched overnight in tryptic soy broth (TSB) at 37 degrees C followed by overnight enrichment in Rappaport-Vassiliadis 10 broth (RV10) at 42 degrees C; (ii) 24 water samples from the third farm were preenriched overnight in 3MC broth at 37 degrees C followed by overnight enrichment in RV10 at 42 degrees C; and (iii) 34 fecal samples from a fourth farm, a finishing farm, were enriched overnight in RV10 at 42 degrees C with no additional enrichment. Following each of the enrichment techniques, samples were subcultured onto modified semisolid Rappaport-Vassiliadis (MSRV) agar prior to transfer to Hektoen Enteric agar plates for the recovery of viable Salmonella bacteria. Presumptive Salmonella isolates were biochemically and serologically confirmed. For the PCR detection of Salmonella, a 1-ml portion was removed from each sample after the first overnight enrichment and the DNA was extracted using a Sepharose CL-6B spin column. Amplicons (457 bp) derived from primers to the invA and invE genes were confirmed as Salmonella specific on ethidium bromide-stained agarose gels by Southern hybridization with a 20-mer oligonucleotide probe specific for the Salmonella invA gene. Neither the standard microbiological method nor the molecular method detected all of the 65 samples that tested positive by both methods or either method alone. Salmonella bacteria were detected by both cultivation and PCR-hybridization in 68% (17 of 25) of the positive samples that were preenriched in TSB, in 73% (11 of 15) of the positive samples preenriched in 3MC broth, and in 24% (6 of 25) of the positive samples enriched in RV10. Agreement between Salmonella detection using cultivation with preenrichment and detection by PCR was 76% using the kappa statistic. However, agreement between Salmonella detection using cultivation without preenrichment and detection by PCR was about 6%; the PCR assay detected 80% (20 of 25) of the 25 positive samples, while Salmonella bacteria were recovered from only 44% (11 of 25) by cultivation. Our results indicate that the PCR-hybridization approach is equivalent to or better than cultivation for detecting Salmonella in swine feces or water samples from swine farms when using the medium combinations evaluated in this study.

Animal Husbandry↗

Sensitivity and specificity of somatic cell count and California Mastitis Test for identifying intramammary infection in early lactation.

Associations between values for the somatic cell count (SCC) or the California Mastitis Test (CMT) and intramammary infection (IMI) were studied in 131 dairy cows from three herds during the first 10 d post-calving. Intramammary infection was defined as the presence of one or two bacterial species in one or both quarter milk samples taken within 12 h of calving and at d 3 postcalving. Quarter milk samples identified IMI in 36% of glands. Values for SCC declined at a significantly faster rate over the first 10 d postcalving in non-infected quarters than in infected quarters. The usefulness of quarter milk SCC and CMT for screening was evaluated by calculating the sensitivity and specificity for various threshold values and days postcalving. A SCC threshold of 100,000 cells/ml for quarter samples evaluated on d 5 postcalving had the maximal sensitivity and specificity for detecting IMI. Evaluation of the CMT samples taken on d 3 postcalving using a threshold reaction of greater than zero had the highest sensitivity and specificity for detecting IMI. With this CMT sampling scheme, the sensitivities for detecting IMI with any pathogen, IMI with a major pathogen, and IMI with a minor pathogen were 56.7, 66.7, and 49.5, respectively. The CMT could have a useful role in dairy herd monitoring programs as a screening test to detect fresh cows with IMI caused by major pathogens.

Animals↗

Neurotransmitter-stimulated ion transport across cultured bovine mammary epithelial cell monolayers.

Bovine mammary epithelial (BME-UV) and myoepithelial (BMM-UV) cell lines were acquired with the goal of developing an in vitro model of mammary epithelia for the study of ion transport. The bovine mammary cell lines were successfully cultured on commercially available permeable supports, and results suggest that mammary epithelial cells, but not myoepithelial cells, form tight junctions necessary to perform a barrier function. Electrogenic ion transport was not observed in basal conditions. Acute exposure to norepinephrine or forskolin caused prototypic increases in short circuit current accompanied by a reduction in transmural resistance indicative of anion secretion through a conductive pathway. Bumetanide and N-(4-methyphenylsulfonyl)-N'-(4-trifluoro-methylphenyl)urea, inhibitors of Na+/K+/Cl- cotransport and cystic fibrosis transmembrane conductance anion channels, respectively, reduced forskolin-stimulated ion transport. Amiloride, an inhibitor of epithelial sodium channels, had no effect on basal or forskolin-stimulated ion transport. However, naturally occurring and synthetic corticosteroids induced the expression of amiloride sensitive current indicative of sodium absorption. Chronic exposure to increased apical ionic strength and/or reduced carbohydrate concentration were associated with reduced transepithelial resistance although forskolin-stimulated ion transport was unaffected. These results demonstrate that neurotransmitters and steroid hormones act directly on bovine mammary epithelial cells to acutely and chronically modulate the volume and composition of their secretions. The in vitro system that we describe can be further exploited to characterize cellular and molecular mechanisms associated with mammary function in health and disease.

Amiloride↗

Escherichia coli O157:H7 in free-ranging deer in Nebraska.

In order to determine the prevalence and distribution of the human pathogen, Escherichia coli O157:H7, in free-ranging deer, hunters were asked to collect and submit fecal samples from deer harvested during a regular firearm season (14-22 November 1998). Prior to the season, 47% of the hunters with permits in the southeastern Nebraska (USA) study area indicated a willingness to participate in the study. Approximately 25% of successful hunters in the area submitted deer fecal samples. Escherichia coli O157:H7 was cultured from four (0.25%) of 1,608 total samples submitted. All of the fecal samples that were properly identified (1,426) and all that were positive for E. coli O157:H7 were from white-tailed deer (Odocoileus virginianus). We were unable to detect a statistically significant geographic distribution pattern of E. coli O157:H7. The presence of E. coli O157:H7 in the feces of free-ranging deer has implications not only for hunters, consumers of venison, and others in contact with deer or deer feces, but also for the development of strategies aimed at reducing and/or controlling this pathogen in water sources and domestic livestock.

Animals↗

Presence of glutamine at position 74 of pocket 4 in the BoLA-DR antigen binding groove is associated with occurrence of clinical mastitis caused by Staphylococcus species.

Potential relationships between amino acid motifs in the antigen binding groove of various alleles of the bovine major histocompatibility complex DR (BoLA-DR) molecule and occurrence of clinical mastitis caused by Staphylococcus species (non-Staphylococcus aureus) were investigated in a case-control study. A significant association (P< or =0.05) was detected between the presence of glutamic acid at position beta 74 and occurrence of mastitis caused by Staphylococcus spp. with a relative risk of 11. This motif is present in BoLA-DRB3.2*22, *23 and *24 alleles. Presence of a positively charged residue (arginine or lysine) at position 13 also showed a tendency (P< o r=0.1) towards an association with a higher risk of clinical mastitis caused by the same bacteria. This motif is present in BoLA-DRB3.2*23 and *8 alleles. Similarly, presence of arginine at position beta 71 (present in alleles *23 and *22) was associated with occurrence of this disease. These positions (beta 13, beta 71 and beta 74) form pocket 4 of the antigen binding groove, which plays an instrumental role in antigen binding and recognition by T-lymphocytes. Thus, it can be concluded that pocket 4 of the BoLA-DR molecule is involved in conferring susceptibility to clinical mastitis caused by Staphylococcus spp.

Alleles↗

Results of a longitudinal study of the prevalence of Escherichia coli O157:H7 on cow-calf farms.

OBJECTIVE: To describe the frequency and distribution of Escherichia coli O157:H7 in the feces and environment of cow-calf herds housed on pasture. SAMPLE POPULATION: Fecal and water samples for 10 cow-calf farms in Kansas. PROCEDURE: Fecal and water samples were obtained monthly throughout a 1-year period (3,152 fecal samples from 2,058 cattle; 199 water samples). Escherichia coli O157:H7 in fecal and water samples was determined, using microbial culture. RESULTS: Escherichia coli O157:H7 was detected in 40 of 3,152 (1.3%) fecal samples, and 40 of 2,058 (1.9%) cattle had > or = 1 sample with E coli. Fecal shedding by specific cattle was transient; none of the cattle had E coli in more than 1 sample. Significant differences were not detected in overall prevalence among farms. However, significant differences were detected in prevalence among sample collection dates. Escherichia coli O157:H7 was detected in 3 of 199 (1.5%) water samples. CONCLUSIONS AND CLINICAL RELEVANCE: Implementing control strategies for E coli O157:H7 at all levels of the cattle industry will decrease the risk of this organism entering the human food chain. Devising effective on-farm strategies to control E coli O157:H7 in cow-calf herds will require an understanding of the epidemiologic characteristics of this pathogen.

Animal Husbandry↗

Use of infrared thermography to detect inflammation caused by contaminated growth promotant ear implants in cattle.

OBJECTIVE: To compare the infrared thermographic appearance of bovine ears that had received contaminated growth promotant implants with ears that had received clean implants and ears without implants. DESIGN: Prospective study. ANIMALS: 32 yearling crossbred beef steers with a mean weight of 322 kg (708 lbs). PROCEDURE: Contaminated (n = 16) and clean (16) implants were placed in the ears of feedlot cattle. Nonimplanted (n = 32) ears served as a within-animal control for thermographic comparisons. Images of rostral and caudal surfaces were obtained during a 21-day period, using an infrared thermal imaging radiometer. Repeated measures ANOVA was used to determine the relationship between mean temperature in a zone on the rostral surface of the ear and at 3 locations (proximal, middle, distal) on the caudal surface of the ear (response variables) with treatment (ears with contaminated implants or clean implants vs control ears with no implants), time (repeated day of measurement), and interactions among these variables. RESULTS: Significant temperature differences existed between ears with contaminated implants and control ears. Temperatures for ears with clean implants were significantly higher than control ears on day 2. At low ambient temperatures when the ears became wet, a greater temperature contrast was detected between ears with contaminated implants and control ears. CONCLUSIONS AND CLINICAL RELEVANCE: Thermal imaging of the ears of feedlot cattle is a noninvasive diagnostic tool that can be used to identify cattle with abscesses caused by contaminated growth-promotant implants.

Administration, Topical↗

Prevalence of Escherichia coli O157:H7 in white-tailed deer sharing rangeland with cattle.

OBJECTIVE: To determine the prevalence of fecal shedding of Escherichia coli O157:H7 in white-tailed deer (Odocoileus virginianus) with access to cattle pastures. DESIGN: Survey study. SAMPLE POPULATION: 212 fecal samples from free ranging white-tailed deer. PROCEDURE: Fresh feces were collected on multiple pastures from 2 farms in north central Kansas between September 1997 and April 1998. Escherichia coli O157:H7 was identified by bacterial culture and DNA-based methods. RESULTS: Escherichia coli O157:H7 was identified in 2.4% (5/212) of white-tailed deer fecal samples. CONCLUSIONS AND CLINICAL RELEVANCE: There is considerable interest in the beef industry in on-farm control of E coli O157:H7 to reduce the risk of this pathogen entering the human food chain. Results of our study suggest that the design of programs for E coli O157:H7 control in domestic livestock on pasture will need to account for fecal shedding in free-ranging deer. In addition, the results have implications for hunters, people consuming venison, and deer-farming enterprises.

Animals↗

Associations of the bovine major histocompatibility complex DRB3 (BoLA-DRB3) with production traits in Canadian dairy cattle.

Associations of two alleles of the bovine major histocompatibility complex DRB3 gene (BoLA-DRB3) with lowered somatic cell score (SCS) and occurrence of disease (BoLA-DRB3.2* 16 and *23, respectively) have previously been documented. The objective of this study was to evaluate potential relationships between BoLA-DRB3 alleles with production traits, namely 305-day milk, milk fat and milk protein yield, in a population of Canadian dairy cattle (Holstein, n = 835 and Jersey, n = 66) over the course of two lactations. No significant associations were detected between BoLA alleles and production traits in Jerseys. In Holsteins, alleles *16 and *23 also did not show associations with production traits but allele *8 was significantly associated with increased 305-day milk, fat and protein yields in the previous lactation (the lactation prior to immunization with a gram negative core antigen vaccine), and with increased protein production in the subsequent (with reference to the time of immunization) lactation. Allele *22 was associated with decreased milk and protein yield in both previous and subsequent lactations. Therefore, it can be concluded that increasing or decreasing the frequency of BoLA alleles *16 and *23 to reduce SCS or increase resistance to mastitis in this population would not have adverse effects on production in this population, and that certain BoLA alleles (*8 and *22) are associated with altered production traits in Canadian Holsteins.

Alleles↗