Occurrence of Mycobacterium avium subsp. paratuberculosis in milk at dairy cattle farms: a systematic review and meta-analysis

Presence of Mycobacterium avium subsp. paratuberculosis (MAP) in milk for human consumption is a concern due to its possible relationship with Crohn's disease in humans. Pasteurization effectively reduces the MAP load by four to five logs, but the efficacy depends on the MAP concentration, which depends on the prevalence among contributing herds and individuals. Considerable variation of MAP in bulk tank milk (BTM) and individual cow's milk (IM) is reported, but factors associated with MAP occurrence in milk at farm level have not been described. This study systematically reviewed published studies aiming at estimating the occurrence of MAP in on-farm BTM and IM by meta-analysis. A total of 692 articles were identified through electronic databases and initially screened using title and abstract. The quality of the 61 potentially relevant articles was assessed using full text and 31 articles were eventually included in the meta-analysis. The apparent prevalence (AP) of MAP in BTM and IM on farm were summarized in relation to strata defined by the test used to identify MAP and the infection status of the herds/animals. There was considerable inconsistency in the reporting, resulting in missing information potentially explaining the dispersion in the estimated AP. The overall AP and 95% confidence intervals based on PCR and culture of MAP were summarized to 0.10 (0.04-0.22) in BTM and 0.20 (0.12-0.32) in IM. Quantifying the MAP load in test-positive milk samples was not possible because very few articles provided quantitative information on individual samples.     

Rehydration fluid temperature affects voluntary drinking in horses dehydrated by furosemide administration and endurance exercise

To determine whether temperature of rehydration fluid influences voluntary rehydration by horses, six 2-3-year-old horses were dehydrated (4-5% body weight loss) by a combination of furosemide administration and 30 km of treadmill exercise. For the initial 5 min following exercise, horses were offered a 0.9% NaCl solution at 10, 20, or 30 degrees C. Subsequently, after washing and cooling out, voluntary intake of water at 10, 20, or 30 degrees C from 20 to 60 min after exercise was measured. Fluid intake (FI) during the first 5 min of recovery was 9.8+/-2.5,12.3+/-2.1 and 9.7+/-2.0L (p>0.05) for saline at 10, 20, and 30 degrees C, respectively. Although not a significant finding, horses offered 0.9% NaCl at 20 degrees C tended to take fewer (p=0.07), longer drinks than when saline at either 10 or 30 degrees C was offered. Between 20 and 60 min of recovery, intake of water at 20 degrees C (7.7+/-0.8L) and 30 degrees C (6.6+/-1.2L) was greater (p<0.05) than that at 10 degrees C (4.9+/-0.5L). Thus, total FI was 14.7+/-2.5,19.9+/-2.5, and 16.3+/-2.4L for rehydration fluids at 10, 20, and 30 degrees C, respectively (p<0.05, value for 20 degrees C water greater than that for 10 degrees C water). Although the amount of metabolic heat transferred to the initial saline drink was correlated with the decrease in core temperature during the initial 5 min of recovery, heat transfer to ingested fluid was most likely responsible for the dissipation of, at most, 5% of the heat generated during endurance exercise. In conclusion, following exercise these dehydrated-normothermic horses voluntary drank the greatest amount of fluid at near ambient (20 degrees C) temperature. Although not determined in this study, greater satiation of thirst by oropharyngeal cooling may have contributed to lesser intake of colder (10 degrees C) fluid.     

Influence of age,sex and herd characteristics on the occurrence of Verocytotoxin-producing Escherichia coli O157 in Danish dairy farms

Verocytotoxin-producing Escherichia coli (VTEC) O157 is an important emerging human pathogen. Cattle are considered to be the main reservoir for VTEC O157. The objectives of this study were to determine the prevalence of VTEC O157 in Danish dairy herds and to investigate the relationship between shedding of VTEC O157 and a number of animal and herd characteristics. Sixty dairy farms were visited once in August-October, and from each herd faecal samples from up to 50 animals were analysed for VTEC O157 by enrichment, immunomagnetic separation (IMS), and plating on selective agar. In total, 2419 animals were sampled, and 3.6% of these excreted VTEC O157. These animals were located on 10 farms (17%). On average, 21% of the sampled animals in the positive herds excreted VTEC O157. Register data, including age, sex, breed, housing conditions and herd composition, were extracted from a database. No influence of herd size or housing conditions was found. A strong effect of age was seen with 2-6-month-old calves as the high-risk age group (8.6% positive) in contrast to calves <2 months (0.7%) and cows (2.4%). There was a non-significant tendency of bull calves to have a higher prevalence than heifers within the age group of 2-6 months. Significantly, more of the herds characterised by having relatively many bull calves or many animals bought into the herd were positive for VTEC O157. Despite the low incidence of human VTEC O157 infections in Denmark, the prevalence in Danish dairy herds was found to be at a similar level as in many other countries.     

Responses to Selection against Clinical Mastitis in the Norwegian Cattle Population

The genetic trend for clinical mastitis was estimated for Norwegian Cattle, a population in which mastitis resistance has been included in the breeding programme since 1978. More than 1.2 million first-lactation daughters, with clinical mastitis records registered from 1978 to 1995, bred by 2043 sires, were analysed with a linear sire model. For bulls born from 1974 to 1990 a flat genetic trend for clinical mastitis was found. However, bull sires born from 1983 onwards, selected on a breeding goal with increased weight on mastitis relative to milk production, showed consistently larger and favourable selection differentials for mastitis relative to bull sires born earlier, which had an average selection differential of approximately zero. This will influence genetic trend in the population through their sons, which were born from 1990 onwards.     

Selection of Nontarget Testing Organisms for ERA of GM Potato with Increased Resistance to Late Blight

The deliberate release of any genetically modified (GM) organism in the European Union requires an environmental risk assessment (ERA) prior to commercialisation, including impact assessment on nontarget organisms. We report from two expert workshops where a newly developed selection procedure for identification of ecologically relevant testing organisms was applied to the case of a GM potato with increased resistance to late blight, planned for cultivation in southern Scandinavia. Species known to contribute to important ecological functions in the receiving environment were selected in a stepwise procedure, to arrive at a practical number of ecologically relevant species that are likely to be exposed to the transgene and suitable for experimental testing. Four ecological functional categories were identified: herbivory and disease transmission, natural enemies, ecological soil processes and pollination. Among these, relevant nontarget species were identified for herbivores and soil living pathogens, natural enemies and decomposers/beneficial soil organisms. Out of a total of 16 herbivores, 17 soil-living pathogens, 49 natural enemies and 14 decomposers/beneficial soil organisms in the initial lists, 8 herbivores, 10 soil-living pathogens, 15 natural enemies and 11 decomposers/beneficial soil organisms were identified as possible testing organisms, based on ecological criteria. These findings are highly relevant for determining the scope and structure of an ERA of this type of GM potato. The selection procedure could not be completed because of insufficient information about tissue- and developmental stage-specific expression levels of the transgenic products for this particular GM potato. Thus, the case study illustrates some of the difficulties and knowledge gaps that limit the relevance and quality of ERA of GM plants.     

Streptococcus equi subsp. zooepidemicus isolates from equine infectious endometritis belong to a distinct genetic group

Streptococcus equi subsp. zooepidemicus is the pathogen most commonly isolated from the uterus of mares. S. zooepidemicus is an opportunistic pathogen and part of the resident flora in the caudal reproductive tract. The aim of this study was to investigate whether a genotypically distinct subpopulation of S. zooepidemicus is associated with endometritis in the mare, by genotyping and comparing uterine S. zooepidemicus strains with isolates from the vagina and clitoral fossa. Mares with (n = 18) or without (n = 11) clinical symptoms of endometritis were included. Uterine samples were obtained using a guarded endometrial biopsy punch, whereas a swab was used to recover samples from the cranial vagina and the clitoral fossa. If S. zooepidemicus was present, up to three colonies were selected from each anatomical location (max. 9 isolates per mare). Bacterial isolates were characterized by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). S. zooepidemicus was isolated from the endometrium of 12 mares. A total of 88 isolates were analyzed by PFGE: 31 from the endometrium, 26 from the cranial vagina and 31 isolates from the clitoral fossa. For MLST 21 isolates were chosen. Results demonstrated a higher genetic similarity of the isolates obtained from infectious endometritis compared to isolates obtained from the caudal reproductive tract. In conclusion, we demonstrate for the first time that a genetically distinct group of S. zooepidemicus is associated with infectious endometritis in the mare.     

In vitro adherence of Staphylococcus pseudintermedius to canine corneocytes is influenced by colonization status of corneocyte donors

The current knowledge of in vitro adherence of Staphylococcus pseudintermedius to canine corneocytes is limited to comparative analyses between strains, staphylococcal species or corneocytes collected from different breeds, body sites and hosts. However, the role played by colonization status of corneocyte donors remains unknown. The aim of this study was to evaluate the adherence properties of commensal S. pseudintermedius strains to corneocytes collected from dogs with different colonization status. For this purpose, corneocytes were collected from five dogs that were classified as persistently colonized (D1 and D2), intermittently colonized (D3 and D4) or non-colonized (D5) on the basis of the results of a previous longitudinal study. Adherence to corneocytes originating from each of the five dogs was assessed by an in vitro adhesion assay using four genetically unrelated strains isolated from the colonized dogs (S1 to S4). Irrespective of their host of origin, all strains adhered significantly better to corneocytes from D1 and D2 than to corneocytes from D3, D4 and D5 (P < 0.0001). The mean count of cells adhering to corneocytes from persistently colonized dogs was on average three times higher than the mean count using corneocytes from the other dogs. A significant difference between strains was only observed for one strain-corneocyte combination (S2-D4), indicating that S. pseudintermedius adherence to corneocytes is driven by host factors and only marginally influenced by strain factors. This finding has important implications for understanding and preventing S. pseudintermedius skin colonization and infection.     

Ascorbic acid and ascorbic acid 6-palmitate induced oxidation in egg yolk dispersions

The oxidation in aqueous dispersions of egg yolk powder and the influence of addition of the proposed antioxidants ascorbic acid and ascorbic acid 6-palmitate indicate that both ascorbic acid and ascorbic acid 6-palmitate propagated the oxidation of egg yolk powder dispersions. Ascorbic acid 6-palmitate was found to be more prooxidative than ascorbic acid. Moreover, it was found that addition of ascorbic acid or ascorbic acid 6-palmitate gave rise to an increase in the amount of free iron Fe(II) in the egg yolk dispersions. It is proposed that ascorbic acid and ascorbic acid 6-palmitate react with the phosvitin-Fe(III) complex found in egg yolk and release Fe(II), which subsequently propagates lipid oxidation. It appears that less oxidation occurs in egg yolk dispersions exposed to high concentrations of peroxy radicals with added ascorbic acid than egg yolk dispersions with added ascorbic acid without exposure to peroxy radicals.     

Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide

A polyamide nucleic acid (PNA) was designed by detaching the deoxyribose phosphate backbone of DNA in a computer model and replacing it with an achiral polyamide backbone. On the basis of this model, oligomers consisting of thymine-linked aminoethylglycyl units were prepared. These oligomers recognize their complementary target in double-stranded DNA by strand displacement. The displacement is made possible by the extraordinarily high stability of the PNA-DNA hybrids. The results show that the backbone of DNA can be replaced by a polyamide, with the resulting oligomer retaining base-specific hybridization.