Effect of Breeding Activity on the Microflora of the External Genitalia and in the Semen of Stallions,and the Relationship Between Micro‐organisms on the Skin and on the External Genitalia

A possible role of breeding activities in the composition of the microbial population in stallions' external genitalia (EG) and the relationship between micro‐organisms colonizing the skin of the abdomen and the ones colonizing the EG have not been studied. In experiment 1, EG microbiological samples were collected from 41 stallions used for both natural cover and semen collection (BST) and from 18 non‐breeding stallions (NBST). A higher (p < 0.05) frequency of isolation of potentially pathogenic species was found for BST. Age did not influence number of micro‐organism species isolated both in BST and NBST. In experiment 2, the microbial content of the EG and semen was compared in 23 BST. Most micro‐organisms isolated from the EG were present in semen, albeit with a numerically lower prevalence. In 7 stallions, six microbial species isolated from semen were absent from the EG cultures, suggesting contamination by the operator. In experiment 3, a numerically higher number of micro‐organism species was isolated from the EG of 31 stallions, than from their skin of the ventral abdomen in contact with the penis or from the skin of the thorax. With the sole exception of Escherichia coli, potentially pathogenic bacteria were only isolated from the EG but not from the skin. Results suggest that breeding activity increased the number of species colonizing the EG; most species isolated from the EG were also found in semen even if with a lower frequency, and additional semen contamination seemed to occur during its manipulation. Many micro‐organism species of the skin were also isolated from the penis, but independently of being or not in contact with the penis, skin did not seem to provide an adequate environment for the growth of potentially pathogenic bacteria that were isolated from EG, with the sole exception for E. coli.     

Characterization of Amaranthus cruentus L. seed proteins by 2-DE and LC/MS–MS: Identification and cloning of a novel late embryogenesis-abundant protein

Amaranth is taking great attention as an important cereal crop that could fulfill food requirements for the growing population, especially in developing countries. However, the protein composition of these seeds is not well known yet. We have used the proteomics tools to characterize amaranth seed proteome. About 400 proteins spots were resolved on two-dimensional gel electrophoresis (2-DE) and protein spots were analyzed by LC-MS/MS (liquid chromatography tandem mass spectrometry). Identified proteins were related to stress and defense responses, metabolic, respiratory, and oxide-reduction processes. One abundant spot was identified as a Late Embryogenesis Abundant (LEA) protein and the gene was cloned and characterized. The AcLEA cDNA contains a 418 bp open reading frame (ORF) encoding a polypeptide of 139 amino acids. Bioinformatics analysis showed that AcLEA belongs to LEAs Group 5. Proteomics is a powerful technique that could be used even in non-sequenced organisms such as amaranth. The obtained information reveals that amaranth seed, beyond the classical seed storage proteins, contains proteins related to protection against stress. The identification of these proteins opens the door to the application of new strategies to improve the quality of amaranth production.     

Development and evaluation of a rapid absorbed enzyme immunoassay test for the diagnosis of Johne''s disease in cattle

An absorbed enzyme immunoassay (EIA) test for Johne's disease in cattle was developed in which absorption of cross-reacting antibodies occurred as a rapid reaction in solution rather than overnight with whole organisms and a subsequent centrifugation step. Total test time was reduced to less than 2 h with a minimum of manipulations. The test was evaluated in cattle herds from Johne's disease-endemic and Johne's disease-free regions of Australia. Specificity was 99.8%. Calculations of sensitivity were affected by the history of the herd under test. However, the EIA detected in excess of 80% of animals before onset of clinical disease and 65% of faecal shedders were EIA positive on, or before, first detection of Mycobacterium paratuberculosis in their faeces. The test should aid epidemiological studies and be a useful tool in the management and control of Johne's disease.     

The prevalence of itchmite, Psorergates ovis, among sheep flocks with a history of fleece derangement

An investigation of sheep flocks in the main sheep raising areas of New South Wales showed that the itchmite Psorergates ovis was frequently associated with fleece derangement. In 26 of the 41 flocks examined, P. ovis was the only ectoparasite detected. P. ovis and the sheep body louse Damalinia ovis, were found in 5 flocks. No external parasites were found on sheep examined from the 10 remaining flocks. The type of fleece derangement most frequently recorded was rubbing which in some cases was combined with areas of chewed fleece. Among flocks, there were positive relations between the prevalence of fleece derangement and prevalence of itchmite or scurf and between itchmite count and mean scurf score. Within flocks, itchmite infested sheep or sheep with scurf had higher prevalences of fleece derangement than sheep on which no mites or no scurf were found. Itchmite infested sheep had a higher prevalence of scurf than those with no detectable mite infestation. There were no significant differences in itchmite populations or fleece derangement between untreated flocks and flocks treated with synthetic pyrethroids, organophosphates or arsenic and rotenone.     

Fungicide dissipation curves in winemaking processes with and without maceration step

The evolution of residual levels of four fungicides (cyprodinil, fludioxonil, pyrimethanil, and quinoxyfen) during the elaboration of three types of wine with maceration (traditional red wine, carbonic maceration red wine, and red wine of long maceration and prefermentation at low temperature) and two types of wine without maceration (rose and white) has been studied. The disappearance curves of each fungicide have been analyzed during the period of each winemaking process (21 days) and during the different enological steps involved in the elaborations. The residual levels of fludioxonil reduce most quickly during the winemaking processes without maceration, whereas the decrease in levels of pyrimethanil was the slowest in practically all cases (with and without maceration). During carbonic maceration winemaking, the decay constant of cyprodinil was greater than that of the other pesticides in all assays (time and steps).     

Evolution of chlorpyrifos, fenarimol, metalaxyl, penconazole, and vinclozolin in red wines elaborated by carbonic maceration of Monastrell grapes

The influence of the different steps involved in the wine-making process on the disappearance of chlorpyrifos, fenarimol, metalaxyl, penconazole, and vinclozolin in red wines elaborated by carbonic maceration of Monastrell grapes was studied. The initial levels of the residues in grapes ranged from 0.28 mg/kg (penconazole) to 1 mg/kg (chlorpyrifos). Ten days after the beginning of maceration, the compound that had decreased least was chlorpyrifos (83% of initial value remaining), whereas metalaxyl, the least persistent of the residues, had decreased to 49% of its initial value. In the free-run juice, on the other hand, the highest percentage remaining corresponded to metalaxyl (10%) and the lowest to chlorpyrifos (0. 1%). After pressing, the percentages of initial values eliminated in pomace varied from 82.7% for chlorpyrifos to 17.7% for metalaxyl, whereas in the press juice the opposite was the case (37% metalaxyl and 2% chlorpyrifos remaining). In finished wine, there were residues of all the pesticides, with the exception of chlorpyrifos, metalaxyl (21%) being the most persistent. The percentages eliminated in the lees varied from 1.5 to 2.5% of the initial value.     

Direct measurement of the current-phase relation of a superfluid 3He-B weak link

Direct measurements of the current-phase relation, I versus Deltaphi, for a weak link coupling two reservoirs of B-phase superfluid helium-3 (3He-B) were made over a wide range of temperatures. The weak link consists of a square array of 100-nanometer-diameter apertures. For temperatures T such that T/Tc >/= 0.6 (where Tc is the superfluid transition temperature), I approximately sin(Deltaphi). At lower temperatures, I(Deltaphi) approaches a straight line. Several remarkable phenomena heretofore inaccessible to superconducting Josephson junctions, including direct observation of quantum oscillations and continuous knowledge of Deltaphi, were also observed.     

Controlling microbial contamination on beef and lamb meat during processing

SUMMARY The microbiological quality of carcases, meat and environmental surfaces was evaluated in commercial boning rooms processing beef and lamb. There was considerable variation in the level of microbial contamination on both carcases and meat, with counts ranging from less than 20 to 10⁸/cm² on carcases and to 2 times 10⁷/cm² on meat. The level of microbial contamination on meat was influenced by the level of carcase contamination at boning and by the boning process itself. Carcase contamination was the major determinant of microbiological quality, as more than 70% of carcases had microbial counts greater than 10³/cm². Cutting boards were a major source for microbial dissemination during boning, particularly when carcase counts were less than 10³/cm². If carcases were heavily contaminated, the contamination of processing surfaces was irrelevant in determining microbial loads on meat. Where carcase contamination was at low to moderate levels, the contribution of the boning process to the contamination on meat assumed increased significance. Under these conditions, improved sanitation of cutting surfaces in the boning room resulted in a significant reduction in microbial contamination on the surface of meat. These results can form the basis for ensuring that improvements made in carcase management before boning, to improve microbiological quality, will be preserved through attention to cutting board hygiene during boning.     

Effectiveness of passive gas-scavenging canisters attached to isoflurane anesthesia systems under standard-use conditions in a laboratory animal facility

Chronic exposure to trace levels of waste anesthetic gases has been linked to higher incidences of neurologic and reproductive dysfunction, hepatic and renal toxicity, and neoplasia in health care professionals. We have shown that low levels of isoflurane emissions are likely in conventional laboratory animal treatment rooms during the use of standard anesthesia delivery systems equipped with activated charcoal canisters for passive gas scavenging. In the present study, we surveyed the effectiveness of canisters (attached to well‐maintained precision isoflurane vaporizers) in current use throughout our AAALAC‐accredited laboratory animal facility. Canisters (Omnicon f/air) had been weighed prior to use and then attached to dual‐loop systems (face mask and induction box circuits) from 1 week to 6 months of service. Isoflurane emissions were measured using a pre‐calibrated, portable infrared spectrophotometer by attaching each canister to the face‐mask circuit, occluding the face mask and closing the stopcock to the induction circuit, and running the system at uniform isoflurane concentration (2%) and oxygen flow rate (1 L minute^?1). Samples were taken in animal procedure rooms (size range, 45–80 m³) in which the air turnover rate ranged between 20 and 30 nonrecirculating changes per hour. Nine of the 60 canisters (15%) in current use were found to have exceeded the manufacturer's recommended use‐life (defined as a weight increase of 50 g). Of these nine, seven canisters did not scavenge isoflurane at all (indicated by emissions greatly exceeding 100 ppm). Isoflurane was not detected in the operator's breathing zone under normal use conditions (i.e. gas directed to both circuits at once). Of the 51 canisters that had not exceeded their rated use‐life, 12 (23.5%) exhausted isoflurane at >2 ppm. Our data show that (i) the potential for exposure to waste isoflurane emissions will depend on the configuration of the delivery system and that (ii) enhanced attention to canister surveillance may be warranted even in well‐run facilities.