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Variable abattoir conditions affect Salmonella enterica prevalence and meat quality in swine and pork.

Research suggests that abattoir holding pens pose significant Salmonella enterica risk to swine immediately preharvest. The goal of this study was to evaluate those factors related to holding that increased the prevalence of S. enterica in swine at slaughter. To accomplish this goal, we focused on holding time and flooring. Our objectives were to (1) compare Salmonella enterica prevalence among pigs held for short (15-45 min) versus long (up to 4 h) periods before slaughter; and (2) determine the impact of flooring (slatted vs. concrete) as it relates to the prevalence of S. enterica. The study consisted of seven repetitions at a large volume (11,000 head/day) Midwest abattoir. Each repetition consisted of one truck load of pigs (n = 170) sorted into one of three groups: (1) animals held for a short time (15-45 min) on solid floors (short-hold); (2) animals held for 4 +/- 0.5 h on slatted floors; and (3) animals held for 4 +/- 0.5 h on solid concrete floors. At slaughter, samples were collected from 30 pigs in each group. Cecal contents (20 mL), feces (20 g), and the ileocecal lymph node were cultured for S. enterica. Additionally, the effect of holding time on meat quality parameters (loin pH at 35 min and 6 h, color, drip loss) was evaluated for the first four replicates. The proportion of S. enterica-positive samples was highest (p < 0.05) in the cecum of pigs held on solid concrete floors (72.4%), and slightly less for pigs held on slatted floors (63.3%). Animals held for less than 45 min before slaughter demonstrated the lowest proportion of S. enterica-positive samples (52.9%). The pig prevalence, as measured by any one of the three samples being positive, was significantly different (p < 0.05) between animals held on solid floors (81%) and those animals held for 45 min or less before slaughter (69%). Meat quality, as measured by multiple parameters, was adversely affected by lack of a rest period. The mean 24-h pH was significantly lower for the short-hold group compared to the other two groups. The mean Minolta L and the drip loss were significantly higher in the short-hold group. From this and other studies, it appears that elimination of the holding process is not feasible S. enterica control option, given current U.S. harvesting systems.

Abattoirs↗

Molecular methods for diagnosis of Entamoeba histolytica in a clinical setting: an overview.

The range of clinical outcomes following Entamoeba histolytica infection is likely to be influenced by the different strains of the parasite already existing in our population. There is a need for developing faster, reliable and reproducible methods for identifying the different strains of E. histolytica. This would have a major impact on the subsequent course of treatment given to patients. In the post-genomic era, different loci of the Entamoeba genome have been targeted for developing suitable probes and genetic markers. This review highlights the development made in this direction and the possibility of using these methods for routine testing of this parasite in clinical samples.

Animals↗

Fecal bacteria and sex hormones in soil and runoff from cropped watersheds amended with poultry litter.

The application of poultry litter to agricultural fields can provide plant nutrients for crops and forage production, but fecal bacteria and the sex hormones estradiol and testosterone are components of litter that can be detrimental to the environment. Our objective was to determine if applications of poultry litter to small watersheds would contribute to the load of fecal bacteria and sex hormones to soil and runoff. We, therefore, investigated the fate and transport of fecal bacteria, estradiol and testosterone from surface applied poultry litter to four small watersheds. Poultry litter was applied to meet the nitrogen requirements of pearl millet (Pennisetum glaucum L.) in 2000 and grain sorghum [Sorgham bicolor (L.) Moench] in 2001. Neither Salmonella nor Campylobacter were detected in the litter but the fecal indicator bacteria were. The average load of total coliforms,Escherichia coli, and fecal enterococci applied with the litter was 12.2, 11.9, and 12.7 log10 cells ha(−1), respectively. The average load of estradiol and testosterone was 3.1 and 0.09 mg ha(-1), respectively.Runoff events first occurred seven months after the first litter application in 2000, and three weeks after the second application in 2001.Only for the 25 July 2001 runoff event three weeks after the second litter application, were the concentrations of total coliforms, E. coli,and fecal enterococci in runoff greater than background concentrations which were on average 5.2, 1.1, and 2.9 log10 MPN 100 ml(−1),respectively [corrected]. Average background levels of total coliforms, fecal enterococci,and E. coli in surface soil were 8.2, 7.9, and 3.5 log (10) cells kg(−1) soil. At the rate of litter application the concentrations of estradiol and testosterone in the litter did not appear to impact the background levels in the soil and runoff. Because concentrations of sex hormones in litter from other broiler operations are known to be greater than in the litter we applied, further study on the connection between concentrations of sex hormones in poultry litter and operational practices is recommended.

Agriculture↗

Comparison of fecal culture and Danish Mix-ELISA for determination of Salmonella enterica subsp. enterica prevalence in growing swine.

In the USA, control of food-borne salmonellosis associated with meat consumption has been predominantly focused at slaughter and processing. It is expected that standards at slaughter and processing will become more stringent, creating pressure to reduce prevalence of Salmonella-positive food animals through on-farm interventions. The aim of this study was to compare traditional fecal culture and the Danish Mix-ELISA (DME) for determination of Salmonella prevalence pre-harvest in swine. In Trial 1, five cohorts of individually identified pigs were longitudinally sampled during the growing period to compare the kinetics of prevalence as estimated by fecal culture and the DME. In Trial 2, the correlation between fecal prevalence and seroprevalence was estimated pre-marketing in 49 groups of pigs. In Trial 1, fecal prevalence and seroprevalence showed similar kinetics, with a tendency of a higher OD% cut-off to more closely approximate fecal prevalence. In Trial 2, correlations between fecal culture and the DME were 0.40, 0.36, 0.43, and 0.43 (p<0.001) for OD% cut-offs > or =10, 20, 30, and 40, respectively. Based on these results, a higher OD% cut-off would be recommended if more approximate estimation of fecal prevalence is desired and longitudinal sampling would be suggested for evaluating the impact of on-farm interventions for Salmonella reduction whether utilizing fecal culture or the DME. Further evaluation of the impact of Salmonella serovar present on farms on seroprevalence and the relationship of on-farm seroprevalence with food safety risk are needed prior to utilizing the DME for pre-harvest Salmonella diagnostics in the US swine herd.

Age Factors↗

Applications of high-pressure liquid chromatography and field desorption mass spectrometry in studies of natural porphyrins and chlorophyll derivatives.

Mixtures of porphyrins derived from natural sources can be readily separated by high-pressure liquid chromatography both analytically and on a preparative scale. A variety of procedures have been developed not only for the esters but also for free acids, and on the analytical scale quantitation is easily achieved by visible absorption. The retention times are largely characteristic of the number of carboxylic acid side chains (or other polar groups) but further information can be obtained by mass spectrometric studies of the various fractions. Field desorption mass spectrometry is particularly useful for this purpose because the emitter wire can be dipped directly into the eluates. The field desorption spectra of porphyrin free acids and esters as well as their metal complexes give essentially molecular ions with little or no fragmentation in most cases, whereas electron-impact mass spectrometry, particularly of free acids, is impeded by the low volatility of porphyrins. Mixtures can also be analysed by field desorption mass spectrometry, and this provides not only a rapid qualitative assessment of the components of a mixture, but also a check on the subsequent chromatographic separations.

Absorption↗

Detection of anthrax spores in endemic regions of northern Canada.

AIMS: To determine the level of anthrax spore contamination in endemic regions of northern Canada between outbreaks. METHODS AND RESULTS: Bacterial endospores were extracted from specimens via flotation and cultured on selective PLET medium. Of 588 environmental specimens collected, 11 (1.9%) contained viable anthrax spores. CONCLUSION: High environmental concentrations of anthrax spores in northern Canada appear limited to scavenger faeces and anthrax carcass sites. Burial and cremation appear equally effective at removing anthrax spores from the immediate environment, though cremation may be improved by re-burning cremation sites containing unburned animal hair. SIGNIFICANCE AND IMPACT OF THE STUDY: This study describes an effective anthrax spore detection system. It provides the first bacteriological evidence that mammalian scavengers can disseminate anthrax spores in northern Canada, and its results may be compared with future environmental studies of untreated anthrax carcass sites to help improve government response plans.

Animals↗

Combined fluorescent antibody assay and viability staining for the assessment of the physiological states of Escherichia coli in seawaters.

AIMS: A comparison of methods that combine the use of immune sera with specific fluorescent probes for testing viability at single cell level was performed in order to estimate different living attributes of Escherichia coli in natural seawater samples. METHODS AND RESULTS: Cell culturability was assayed by plate method, respiratory activity and membrane integrity were determined by an indirect fluorescent antibody assay, combined with 5-cyano-2, 3 ditolyl tetrazolium chloride and propidium iodide, respectively. Results showed the coexistence of different physiological states within the E. coli population, of which a large fraction (46%) of cells was actively respiring. CONCLUSIONS: The methodological approach used offer interesting perspectives in water pollution monitoring, particularly when the differentiation between dead and living E. coli cells is required for a more precise assessment of the bacteriological quality of seawaters. SIGNIFICANCE AND IMPACT OF THE STUDY: The study suggests the importance of knowledge of the viability status of faecal bacteria in aquatic environments as a fundamental issue for the preservation of public health; the availability of rapid analytical procedures for this purpose may find significant applications in the evaluation of the sanitary risk consequent to water use.

Bacteriological Techniques↗

Detection and identification by real time PCR/RFLP analyses of Cryptosporidium species from human faeces.

AIMS: To detect a wide range of Cryptosporidium species from human faeces by analysis of the Cryptosporidium oocyst wall protein gene by PCR. METHODS AND RESULTS: The nested-assay comprised an initial amplification using a conventional thermocycler followed by real time PCR using a LightCycler with SYBR Green I for the characterization of the amplicons. The technique uses four sets of primers composed of five to six oligonucleotides with one to six base differences corresponding to the inter-species sequence differences of the gene fragment. Restriction fragment length polymorphism analysis identified Cryptosporidium hominis and C. parvum. The assay was evaluated using DNA extracted from purified material and faecal specimens containing a range of potential pathogens (including Cryptosporidium). The assay was specific, sensitive, reproducible and rapid. CONCLUSIONS: This unique technique enables the rapid detection of a range of polymorphic COWP gene sequences directly from faeces using real time PCR. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates a novel approach to identification of Cryptosporidium species and the identification of C. hominis and C. parvum. The technique may be especially useful for the analysis of environmental samples which are likely to contain heterogeneous mixtures of Cryptosporidium species.

Animals↗

Schistosoma mansoni in infants (aged < 3 years) along the Ugandan shoreline of Lake Victoria.

In two complementary epidemiological surveys of villages on the Ugandan shoreline of Lake Victoria, the putative occurrence of intestinal schistosomiasis in the local infants (children aged < 3 years) was investigated. When, during the first survey, 136 mother-and-infant pairs from a total of 12 villages were studied, only 7% of the infants but 45% of the mothers were found to be egg-patent for Schistosoma mansoni infection. The use of dipstick tests for urine-circulating cathodic antigen indicated, however, a much higher prevalence, of approximately 40%, among the infants. In the second survey, urine samples and multiple, not single, stool samples were collected from another 19 mother-and-infant pairs in two of the 12 study villages (Bugoto and Bwondha), and a standardized questionnaire was implemented. The prevalence of egg-patent infection was then found to be markedly higher in the study infants from Bugoto (86%) than in those from Bwondha (25%). A greater level of mother-and-infant water contact, a higher abundance of (infected) Biomphalaria choanomphala, and an unusual lakeshore topology may explain why S. mansoni infection was so much more common in the Bugoto subjects than in the Bwondha. All but one of the infants studied in the second survey were found to be anaemic (with <110 g haemoglobin/litre). Taken together, these children were less likely to be found infected with hookworm (16%), Hymenolepis nana (11%) or Trichuris trichiura (5%) than with S. mansoni (47%). Infection with the parasites causing intestinal schistosomiasis can be common among the infants living in these lakeshore villages. Although the immediate and later-life clinical impacts of such infection have yet to be elucidated, such infants would probably benefit from regular de-worming. Mothers should be strongly encouraged to visit the nearest health-services clinic, with their infants, for any necessary anthelmintic treatment.

Animals↗

[Acute diarrhea: stool water loss in hospitalized infants and its correlation with etiologic agents and lactose content in the diet].

Forty weaned male infants were studied during their first year of life, all hospitalized with acute diarrhea in the Gastroenterology and Metabolism Unit of the Hospital "Umberto I", São Paulo, SP, Brazil. We evaluated and quantified water fecal losses, employing the metabolic bed technique, relating the feeding formula employed with the different causal enteropathogenic agents. 67.5% of the studied infants were under six months and 40% under three months of age. Two groups were randomly assembled to receive, lactose or lactose free feeding formulae. Twenty one patients received a lactose-containing formula (Ninho 10%) and the other 19 children were fed caseine (Portagen) formulae. According to coproculture results and identification of enteropathogenic agents, we divided the studied infants relating feeding formula with the presence or absence of the enteropathogenic Escherichia coli (EPEC): I-13 with positive coproculture for EPEC and diets which included lactose--(L/EPEC+); II--eight with negative coproculture for EPEC and diets which included lactose--(L/EPEC-); III--seven with positive coproculture for EPEC and lactose free diets--(G/EPEC+); IV--12 with negative coproculture for EPEC and lactose free diets. (G/EPEC-). The most frequently isolated agent at coproculture was EPEC, in 20 of the cases (50%), followed by Campylobacter (7.5%). It was also possible to observe that the frequencies of EIEC, Salmonella and Rotavirus were all equal (2.5%). Mixed infections occurred only between EPEC and EIEC, registering a frequency of 5%. The EIEC samples, associated to EPEC 0111 were serotyped as 0 28 ac: H- and 0 152:H-. The use of metabolic bed made the evaluation of fecal volumes possible by a simple and quick technique, thus allowing a closer clinical monitoring, as well as a more reliable evaluation of the patients hospitalized with acute diarrhea. Average acceptance volumes of the formulae--either with or without lactose--were always below the amount recommended by FAO/WHO (100 kcal/day) which shows the impact of acute diarrhea on the decrease of food intake. The average volumes of watery fecal losses found among any of the studied subgroups may be considered quite relevant when compared to standard values. Especially within the L/EPEC+ group fecal losses, both on the first day (83.56 ml/kg/day) and, mainly, on the second (119.44 ml/kg/day) reached exceedingly high levels indicating a disastrous association between the presence of EPEC in the small intestine and lactose offer in the diet. Thus, the results show that there exists a positive and significant association between poor lactose absorption and the presence of EPEC in the feces. WHO's recommendation proposing the use of diluted cow milk, in universal and indiscriminate administration, in the two first days of the disease, may represent a risk factor, not only for malnutrition, but also for the survival rates of children with severe diarrhea, especially those under six months of age and hospitalized with EPEC enteroinfection.

Acute Disease↗

Immune responses to Toxoplasma gondii in the gut.

Toxoplasma gondii is an important cause of disease, in pregnant women, newborns and immunocompromised hosts. Infection with T. gondii naturally occurs through ingestion of raw or undercooked meat containing cysts or through contact with cat feces containing oocysts. The intestine thus appears to be the site of invasion of the parasite. Since acute infection in immunocompetent hosts usually goes unnoticed, no information is available regarding the involvement of the intestine during acute infection in humans. However, in murine models of the disease the acute phase of infection has not been investigated in detail as well. In this report, an overview of the current knowledge regarding the initial phase of infection with T. gondii is given. Results of studies focussing on invasion and dissemination of the parasite and the immune responses elicited in the intestine are reviewed. The impact on the induction of local immune responses of the genetic background and sex of mice as well as the role of different doses and strains of the parasite are discussed. Finally, reports on the development of intestinal pathology in different animal species and the involvement of the intestine in human disease are reviewed.

Animals↗

Geographical variation in antibiotic resistance profiles of Escherichia coli isolated from swine, poultry, beef and dairy cattle farm water retention ponds in Florida.

AIMS: The aim of this study was to assess geographical variation in multiple antibiotic resistance (MAR) profiles of livestock Escherichia coli as well as to evaluate the ability of MAR profiles to differentiate sources of faecal pollution. METHODS AND RESULTS: More than 2000 E. coli isolates were collected from water retention ponds and manure of swine, poultry, beef and dairy farms in south, central and north Florida, and analysed for MAR using nine antibiotics. There were significant differences in antibiotic resistance of E. coli by season and livestock type for more than one antibiotic, but regional differences were significant only for ampicillin. Over the three regions, discriminant analysis using MAR profiles correctly classified 27% of swine, 49% of poultry, 56% of beef and 51% of dairy isolates. CONCLUSIONS: Regional variations in MAR combined with moderate discrimination success suggest that MAR profiles of E. coli may only be marginally successful in identifying sources of faecal pollution. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the existence of regional and seasonal differences in MAR profiles as well as the limited ability of MAR profiles to discriminate among livestock sources.

Agriculture↗

Nitrate and vitamin C from fruits and vegetables: impact of intake variations on nitrate and nitrite excretions of humans.

The objective of the study was to determine nitrate + nitrite excretions of human subjects fed variable amounts of nitrates and nitrites and vitamin C from fruits and vegetables. During four, randomly-arranged experimental periods of seven days each, the 12 apparently healthy, adult human subjects consumed laboratory controlled, constant, diets which were systematically varied in kinds of fruits and vegetable to provide the four following variations: 414 mg nitrate + nitrite and 23 mg vitamin C, 412 mg nitrate + nitrite and 177 mg vitamin C, 23 mg nitrate + nitrite and 39 mg vitamin C, and 21 mg nitrate + nitrite and 193 mg vitamin C per subject per day, respectively. Subjects made complete collections of urine and stools throughout the study. Regardless of type of experimental diet fed, no nitrates and nitrites were detected in the feces. Urinary excretion of nitrate + nitrite was significantly greater at the higher levels of nitrate + nitrite intake than at the lower intake levels. Increased intake of vitamin C at either level of nitrate + nitrite intake resulted in apparent decreased urinary excretions of nitrite + nitrate.

Adult↗

Top dressing of feed with desiccated chlamydospores of Duddingtonia flagrans for biological control of the pre-parasitic stages of ovine Haemonchus contortus.

Feeding trials were conducted with stall-fed sheep parasitized with Haemonchus contortus. For 10 days they were offered 250 g of a concentrate feed that had been top-dressed with desiccated chlamydospores of Duddingtonia flagrans at 1 x 10(5), 5 x 10(5), 1 x 10(6) or 2 x 10(6) chlamydospores/kg body weight. Pooled faeces from each group on day 7 of spore feeding were spread on different pasture plots. On day 28 after the start of spore feeding, further pooled faeces from each group were spread on the same plots. The larval burdens on the plots were monitored for 2 months and the larval harvest from in vitro faecal cultures were monitored regularly. The application of 1 x 10(6) or more spores/kg body weight virtually eliminated larvae from both the pasture and the faecal cultures. The application of as few as 1 x 10(5) spores/kg body weight had a profound impact on larval recovery. The effect persisted while the spores were being fed but not for more than 4 days following discontinuation of spore feeding. Top dressing supplementary feed with dried chlamydospores offers a potential way of using D. flagrans for biological control of the pre-parasitic stages of H. contortus.

Acetyltransferases↗

Carbon source utilization profiles as a method to identify sources of faecal pollution in water.

AIMS: Carbon source utilization profiles as a phenotypic fingerprinting methodology for determining sources of faecal pollution in water were evaluated. METHODS AND RESULTS: Three hundred and sixty-five Enterococcus isolates were collected from known faecal sources in four different geographical regions and were identified to species with the commercial Biolog system. Discriminant analysis (DA) was used to identify the substrate-containing wells that best classified the 365 isolates by source. By using 30 of the 95 wells for the analysis, the average rate of correct classification (ARCC) by source was 92.7% for a human vs non-human two-way classification when isolates from all regions were combined into one library. Corresponding ARCCs for other classification schemes were 81.9% for a four-way classification of human vs livestock vs wildlife vs domestic pets, and 85.7% for a three-way classification without human isolates. When three individual libraries were made based on classification of sources within Enterococcus species, the ARCC was 95.3% for the Ent. faecalis library, 95.8% for the Ent. gallinarum library and 94.7% for the Ent. mundtii library. Thirty Enterococcus isolates (unknown sources) were obtained from each of three stream sites where a specific source of pollution was apparent; 90.0% of the isolates from a human-suspected source were classified as human, 86.6% were classified as livestock from a livestock-suspected site, and 93.3% were classified as wildlife from a wildlife-suspected site. CONCLUSIONS: Phenotypic fingerprinting with carbon source utilization profiles provided levels of correct classification by sources from an Enterococcus library that were in the upper range of those reported in the literature. ARCCs for three Enterococcus species-specific libraries were very high and may be the best approach for further developing this concept and methodology. SIGNIFICANCE ANC IMPACT OF THE STUDY: The results, based on a modest Enterococcus library and a preliminary field validation test, demonstrated the potential for carbon source utilization profiles to be employed as a phenotypic method for determining sources of faecal pollution in water.

Animals↗

Effect of dalbavancin on the normal intestinal microflora.

OBJECTIVES: Dalbavancin is a new lipoglycopeptide antibiotic active in vitro against most Gram-positive bacteria. It is administered parenterally as a weekly regimen, is eliminated both in urine and faeces, and has t(1/2) in plasma of 8.5 days. Investigating the impact of antibiotics on endogenous microflora is important since alteration of the balance may facilitate colonization by new potentially pathogenic strains or enable microorganisms in the normal flora to develop resistance. The purpose of the present study was to investigate the effect of administration of dalbavancin on the intestinal flora of healthy subjects. METHODS: Six women and six men, 18-40 years, received a single 30 min intravenous infusion of 1 g dalbavancin. Plasma and faeces were collected over several weeks for determination of dalbavancin concentration and analysis of faecal flora. Faecal specimens were cultured on non-selective and selective media. Different colony types were counted, isolated in pure culture and identified to genus level. All new colonizing bacteria were tested for susceptibility to dalbavancin. RESULTS: Plasma dalbavancin concentrations at 2, 21 and 60 days after administration were 35.8-208.7, 3.9-22.1 and 0.5-2.9 mg/L, respectively. The faecal concentrations of dalbavancin were 6.8-73.4 mg/kg on day 5 and 7.4-26.4 mg/kg on day 14. Dalbavancin was not detectable in faeces on day 60. There was some impact on numbers of enterococci and Escherichia coli and no changes in numbers of lactobacilli, clostridia and bacteroides. No Clostridium difficile strains were recovered. No new colonizing aerobic and anaerobic bacteria resistant to dalbavancin were found. CONCLUSIONS: Dalbavancin has no major ecological effect on the human normal intestinal microflora.

Adolescent↗

Effect of supplementing corn- or barley-based feedlot diets with canola oil on faecal shedding of Escherichia coli O157:H7 by steers.

AIMS: This study was conducted to evaluate the effect of supplementing barley- or corn-based diets with canola oil on faecal shedding of Escherichia coli O157:H7 by experimentally inoculated feedlot cattle. METHODS AND RESULTS: Four groups of yearling steers fed on barley- or corn-based feedlot diets containing 0% (BA; CO) or 6% canola oil (BA-O; CO-O) were inoculated with 10(10) CFU of a mixture of four nalidixic acid-resistant strains of E. coli O157:H7. The inoculated strains were tracked in oral (mouth swab) and environmental (water, water bowl interface, feed, faecal pat) samples by enrichment and immunomagnetic separation (IMS) for 12 weeks, and in rectally collected faecal samples for 23 weeks (enumeration by dilution plating for 12 weeks; detection by IMS for a further 11 weeks). Levels of E. coli O157:H7 shed in faecal samples over the course of the enumeration period were similar (P = 0.14) among treatments. Disappearance of the inoculated strains from faeces was more rapid (P = 0.009) with barley than with corn, but shedding levels at the end of the enumeration period were similar (P = 0.21) across grain types. Canola oil supplementation did not affect (P = 0.71) the rate of disappearance of E. coli O157:H7 from faeces. The numbers of steers culture positive for E. coli O157:H7 during the enumeration period were similar (P = 0.57) among treatments. During the 11-week detection period, however, more (P < 0.001) steers were E. coli O157:H7-positive in the BA group (15/64) than in BA-O (two of 64), CO (two of 56), or CO-O (one of 56). The organism was present in two of 48 water samples (both CO-O), one of 48 water bowl swabs (BA-O), four of 48 feed samples (two of 12 BA; two of 12 CO-O), 30 of 48 pen floor faecal pat samples, and 296 of 540 mouth swabs (81/144 BA, 80/144 BA-O, 74/126 CO and 61/126 CO-O). CONCLUSION: Supplementing corn or barley-based diets with canola oil did not affect shedding of E. coli O157:H7 by feedlot cattle. SIGNIFICANCE AND IMPACT OF THE STUDY: High-shedding individuals (i.e. 'super shedders') may be responsible for disseminating E. coli O157:H7 among penmates. Faeces on pen floors appears to be a more significant source of infection than are feed or drinking water.

Animal Feed↗

Detection of Mycobacterium avium subspecies paratuberculosis in Swiss dairy cattle by real-time PCR and culture: a comparison of the two assays.

AIMS: To compare the two different diagnostic assays for the detection of Mycobacterium avium ssp. paratuberculosis, the aetiological agent of paratuberculosis. METHODS AND RESULTS: Faecal samples were derived from 310 cows, representing 13 commercial dairy herds in various locations in Switzerland with expected increased risk because of a past history of disease. Detection assays for M. avium ssp. paratuberculosis were culture (gold standard) and a newly designed real-time PCR. Real-time PCR identified 31 of 310 animals as positive within this risk population whereas culture identified 20 positive animals. The specificity of real-time PCR was confirmed by DNA sequencing of the PCR product. Depending on the test used, the paratuberculosis prevalence in our tested risk population ranged from 6.5 to 10%. CONCLUSIONS: Real-time PCR and culture data were in good agreement, and real-time PCR generates data in a short time in contrast to culture. SIGNIFICANCE AND IMPACT OF THE STUDY: We consider real-time PCR as a suitable alternative method to culture for the detection of M. avium ssp. paratuberculosis in a national surveillance programme.

Animals↗