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New sensor technologies enable a valid recording of resting behaviour on pasture. The behaviour of dairy cows influences animal welfare and health, productivity as well as economic parameters. In comparison to freestall housing the resting periods of grazing cows can be postponed or the length of the period can be limited. In the present study, changes in the resting time of dairy cows during the change from freestall housing ("Stall") to grazing ("Weideumstellung") as well as during the subsequent pasture period ("Vollweide") in two experiments (2018 and 2019) at different continuously grazed pasture sward height (AWH) groups. In the stable and pasture conversion phase 18 and 15 animals were kept together and fed uniformly in experiment 1 and 2 respectively. The resting behaviour was recorded during the stable period on the last 9 days before the grazing period started. The subsequent pasture conversion period comprised the first 9 (experiment 1) and 13 (experiment 2) grazing days. Before the subsequent full grazing period started, the cows were divided equally between the two grazing groups in each experiment and then kept on continuous grazed pasture in different sward height groups. The AWH were measured with a rising plate pasture meter. In experiment 1 the AWH groups "kurz" (5.4 cm ±0.15) and "mittel" (6.6 cm ±0.13) and in experiment 2 "mittel" (6.1 cm ±0.87) and "lang" (7.6 cm ±0.95) were compared respectively. During the full grazing period the animals received only 1.4 kg DM of concentrate plus minerals as supplementary feed. In experiment 1, the resting data set included the full grazing weeks 8, 10 and 11 and in experiment 2 the weeks 4, 5, 6, 8 and 12. The resting parameters were recorded with the HOBO Pendant G data logger at a measuring interval of 30 seconds and the data were evaluated with a mixed model. In both experiments the longest daily resting times were recorded during the stable periods and the shortest within the full grazing periods. In experiment 1 the resting time decreased from 11.4 hours per day in the stable period to 10.2 hours in the pasture conversion period and 7.2 to 8.1 hours per day in the full grazing period in the AWH groups "kurz" and "mittel" respectively. In experiment 2 the daily resting times were 11.1 hours for "Stall", 9.7 for "Weideumstellung" as well as 8.5 for AWH group "mittel" and 9.0 hours for group "lang". The average duration of resting (73-87 minutes/period) did not differ significantly between the experimental periods or the AWH groups, but during the full grazing periods, the animals restricted the number of lying phases per day. In comparison to results from the literature, the daily resting times at short AWH can be classi fied as low. Although the available data do not yet allow conclusions to be drawn about reduced animal welfare, measures should be applied in animal and pasture management that allow dairy cow's sufficient time to lie down. [ABSTRACT FROM AUTHOR]

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Ciprofloxacin, azlocillin, ceftizoxime, and amikacin were studied alone and in combination against six Enterobacteriaceae and six strains of Pseudomonas aeruginosa in an infected chamber model in rabbits simulating a closed space infection. In-vivo results were compared with in-vitro tests of inhibition, killing and synergy. Ciprofloxacin was the most effective single agent, with efficacy against five of the six Enterobacteriaceae when used in low doses, and two of the six pseudomonads when used in high doses. The development of in-vitro resistance to ciprofloxacin was observed during therapy in strains which failed to be eradicated. Ciprofloxacin and azlocillin together was the most effective regimen, with efficacy against eleven of the twelve strains. Synergy, as determined by chequerboard testing, did not correlate with in-vivo outcome. Unlike mezlocillin, azlocillin, ceftizoxime or amikacin, MIC testing of ciprofloxacin was predictive of in-vivo success. [ABSTRACT FROM AUTHOR]

A study on the importance of the route of antibiotic administration on achievable extravascular fluid drug concentration was performed in a rabbit model. Cefazolin was given either by repeated intramuscular injection or by constant intravenous infusion for 12 to 26 h. Cefazolin penetration into a high and low protein extravascular space was measured by sampling subcutaneous Visking chambers filled with serum or saline. Application of pharmacokinetic principles to equilibrium conditions allowed accurate prediction of the extravascular fluid concentrations. [ABSTRACT FROM PUBLISHER]

Abstract Background Oral keratinocytes on the mucosal surface are frequently exposed to HIV-1 through contact with infected sexual partners or nursing mothers. To determine the plausibility that oral keratinocytes are primary targets of HIV-1, we tested the hypothesis that HIV-1 infects oral keratinocytes in a restricted manner. Results To study the fate of HIV-1, immortalized oral keratinocytes (OKF6/TERT-2; TERT-2 cells) were characterized for the fate of HIV-specific RNA and DNA. At 6 h post inoculation with X4 or R5-tropic HIV-1, HIV-1gag RNA was detected maximally within TERT-2 cells. Reverse transcriptase activity in TERT-2 cells was confirmed by VSV-G-mediated infection with HIV-NL4-3Δenv-EGFP. AZT inhibited EGFP expression in a dose-dependent manner, suggesting that viral replication can be supported if receptors are bypassed. Within 3 h post inoculation, integrated HIV-1 DNA was detected in TERT-2 cell nuclei and persisted after subculture. Multiply spliced and unspliced HIV-1 mRNAs were not detectable up to 72 h post inoculation, suggesting that HIV replication may abort and that infection is non-productive. Within 48 h post inoculation, however, virus harbored by CD4 negative TERT-2 cells trans infected co-cultured peripheral blood mononuclear cells (PBMCs) or MOLT4 cells (CD4+ CCR5+) by direct cell-to-cell transfer or by releasing low levels of infectious virions. Primary tonsil epithelial cells also trans infected HIV-1 to permissive cells in a donor-specific manner. Conclusion Oral keratinocytes appear, therefore, to support stable non-replicative integration, while harboring and transmitting infectious X4- or R5-tropic HIV-1 to permissive cells for up to 48 h.

Despite their relatively large size, Icelandic glaciers, and their organic carbon (OC) fluxes, have not been explicitly considered in current global glacial OC flux calculations. Most global glacial OC estimates are based on limited individual flux estimates, often determined during the melt season, rarely accounting for the seasonal and diurnal variability of glacial dissolved organic matter (DOM). Using an annual dataset of 25 Icelandic glaciers (and their glacial streams) we investigate DOM concentration and composition, calculating an estimate for downstream OC fluxes from Icelandic glaciers, considering diurnal and seasonal variability. DOM source and composition distinctly changed from a terrestrial character toward a more proteinaceous character as melt increased, both on a seasonal and diurnal basis, likely reflecting the flow path of the meltwater. While DOC concentration did not change on a diurnal basis, DOM composition was more labile in the afternoons, possibly indicating photochemical or biological transformation processes. Overall, the glacial streams predominantly acted as CO 2 sinks. However, higher DOC concentrations, along with contributions of more proteinaceous DOM in proglacial streams, led to a decrease in the uptake potential for CO 2 . Finally, we estimated an export flux of 0.0026 ± 0.0029 Gg C yr -1  km -2 of DOC, and 0.011 ± 0.007 Tg C yr -1  km -2 of POC, from Icelandic glaciers. We reveal larger than previously assumed DOC and POC fluxes from Icelandic glaciers, with higher-than-global-average areal fluxes (~3 % and 9 % of global glacial C flux respectively). Our findings underscore the importance of revising current global estimates to include the not-fully-accounted-for contribution of Icelandic, and other glaciers. This is particularly important considering ongoing climatic changes will likely affect glacial meltwater discharge and sources, leading to altered DOM composition and DOC concentration, having potentially considerable consequences for glacial OC export and CO 2 uptake potentials of glacial streams., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

In aquatic ecosystems, dissolved organic matter (DOM) composition is driven by land use, microbial activity, and seasonal variation in hydrology and water temperature, and, in turn, its microbial bioavailability is expected to vary due to differences in its composition. It is commonly assumed that DOM of terrestrial origin is resistant to microbial activity because it is composed of more complex aromatic compounds. However, the effect of DOM sources on the microbial reworking and degradation of the DOM pool remains debated. We performed laboratory incubation experiments to examine how temporal changes in DOM composition influence its microbial biodegradability in two contrasting streams (agricultural and forested) in southern Ontario, Canada. Despite a more allochthonous-like DOM signature in the forest stream and a more autochthonous-like DOM signature in the agriculture stream, we found that biodegradation and production of DOC were the same in both streams and synchronous throughout the sampling period. However, the initial DOM composition impacted how the DOM pool changed upon degradation. During the incubations, both autochthonous-like and allochthonous-like fractions of the DOM pool increased. We also found that a greater change in DOM composition during the incubations induced higher degradation of carbon. Finally, temporal variation in DOC biodegradation and production over time or across streams was not related to DOM composition, although there was a significant relationship between BDOC and nutrient concentrations in the agriculture stream. This observation potentially challenges the notion that DOM origin predicts its bioavailability and suggests that broad environmental factors shape DOC consumption and production in aquatic ecosystems. More research is needed to better understand the drivers of microbial biodegradability in streams, as this ultimately determines the fate of DOM in aquatic ecosystems., (Copyright © 2020 Kadjeski, Fasching and Xenopoulos.)

Human activities can alter aquatic ecosystems through the input of nutrients and carbon, but there is increasing evidence that these pressures induce nonlinear ecological responses. Nonlinear relationships can contain breakpoints where there is an unexpected change in an ecological response to an environmental driver, which may result in ecological regime shifts. We investigated the occurrence of nonlinearity and breakpoints in relationships between total dissolved nitrogen (TDN), total dissolved phosphorus (TDP), and total dissolved carbon (DOC) concentrations and ecological responses in streams with varying land uses. We calculated breakpoints using piecewise regression, two dimensional Kolmogorov-Smirnov (2DKS), and significant zero crossings (SiZer) methods. We found nonlinearity was common, occurring in half of all analyses, with some evidence of multiple breakpoints. Linearity, by contrast, occurred in less than 14% of cases, on average. Breakpoints were related to land use gradients, with 34-43% agricultural cover associated with DOC and TDN breakpoints, and 15% wetland and 9.5% urban land associated with DOC and nutrient breakpoints, respectively. While these breakpoints are likely specific to our study area, our study contributes to the growing literature of the prevalence and location of ecological breakpoints in streams, providing watershed managers potential criteria for catchment land use thresholds.

Streams and rivers transport dissolved organic matter (DOM) from the terrestrial environment to downstream ecosystems. In light of climate and global change it is crucial to understand the temporal dynamics of DOM concentration and composition, and its export fluxes from headwaters to larger downstream ecosystems. We monitored DOM concentration and composition based on a diurnal sampling design for 3 years in an Alpine headwater stream. We found hydrologic variability to control DOM composition and the coupling of DOM dynamics in the streamwater and the hyporheic zone. High-flow events increased DOM inputs from terrestrial sources (as indicated by the contributions of humic- and fulvic-like fluorescence), while summer baseflow enhanced the autochthonous imprint of DOM. Diurnal and seasonal patterns of DOM composition were likely induced by biological processes linked to temperature and photosynthetic active radiation (PAR). Floods frequently interrupted diurnal and seasonal patterns of DOM, which led to a decoupling of streamwater and hyporheic water DOM composition and delivery of aromatic and humic-like DOM to the streamwater. Accordingly, DOM export fluxes were largely of terrigenous origin as indicated by optical properties. Our study highlights the relevance of hydrologic and seasonal dynamics for the origin, composition and fluxes of DOM in an Alpine headwater stream.

Glaciers harbour diverse microorganisms, which upon ice melt can be released downstream. In glacier-fed streams microorganisms can attach to stones or sediments to form benthic biofilms. We used 454-pyrosequencing to explore the bulk (16S rDNA) and putatively active (16S rRNA) microbial communities of stone and sediment biofilms across 26 glacier-fed streams. We found differences in community composition between bulk and active communities among streams and a stronger congruence between biofilm types. Relative abundances of rRNA and rDNA were positively correlated across different taxa and taxonomic levels, but at lower taxonomic levels, the higher abundance in either the active or the bulk communities became more apparent. Here, environmental variables played a minor role in structuring active communities. However, we found a large number of rare taxa with higher relative abundances in rRNA compared with rDNA. This suggests that rare taxa contribute disproportionately to microbial community dynamics in glacier-fed streams. Our findings propose that high community turnover, where taxa repeatedly enter and leave the 'seed bank', contributes to the maintenance of microbial biodiversity in harsh ecosystems with continuous environmental perturbations, such as glacier-fed streams., (© 2014 The Authors. Environmental Microbiology published by Society for Applied Microbiology and John Wiley & Sons Ltd.)

Streams receive substantial terrestrial deliveries of dissolved organic matter (DOM). The chromophoric (CDOM) fraction of terrestrial deliveries confers the brown colour to streamwater, often understood as browning, and plays a central role in aquatic photochemistry and is generally considered resistant to microbial metabolism. To assess the relevance of terrigenous DOM for carbon fluxes mediated by stream microorganisms, we determined the bioavailable fraction of DOM and microbial carbon use efficiency (CUE), and related these measures to partial pressure of CO2 in headwater streams spanning across a browning gradient. Fluorescence and absorbance analyses revealed high molecular weight and aromaticity, and elevated contributions from humic-like components to characterize terrestrial CDOM. We found that microorganisms metabolized this material at the cost of low CUE and shifted its composition (from fluorescence and absorbance) towards less aromatic and low-molecular weight compounds. Respiration (from CUE) was related to CO2 supersaturation in streams and this relationship was modulated by DOM composition. Our findings imply that terrigenous DOM is respired by microorganisms rather than incorporated into their biomass, and that this channelizes terrigenous carbon to the pool of CO2 potentially outgassing from streams into the atmosphere. This finding may gain relevance as major terrigenous carbon stores become mobilized and browning progresses.

Rad54 is a dsDNA-dependent ATPase that translocates on duplex DNA. Its ATPase function is essential for homologous recombination, a pathway critical for meiotic chromosome segregation, repair of complex DNA damage, and recovery of stalled or broken replication forks. In recombination, Rad54 cooperates with Rad51 protein and is required to dissociate Rad51 from heteroduplex DNA to allow access by DNA polymerases for recombination-associated DNA synthesis. Sequence analysis revealed that Rad54 contains a perfect match to the consensus PIP box sequence, a widely spread PCNA interaction motif. Indeed, Rad54 interacts directly with PCNA, but this interaction is not mediated by the Rad54 PIP box-like sequence. This sequence is located as an extension of motif III of the Rad54 motor domain and is essential for full Rad54 ATPase activity. Mutations in this motif render Rad54 non-functional in vivo and severely compromise its activities in vitro. Further analysis demonstrated that such mutations affect dsDNA binding, consistent with the location of this sequence motif on the surface of the cleft formed by two RecA-like domains, which likely forms the dsDNA binding site of Rad54. Our study identified a novel sequence motif critical for Rad54 function and showed that even perfect matches to the PIP box consensus may not necessarily identify PCNA interaction sites.