Conservation of feline semen. Part I: cooling and freezing protocols

There has been increased interest recently in the conservation of wild felids and preservation of valuable cat breeds. Assisted reproduction, by means of artificial insemination (AI), is an important tool for developing breeding programs for conservation. Optimal use of AI requires accurate data on semen conservation protocols and its long-term storage/survival. In this paper, semen cooling and freezing processes are described, with special emphasis on the results obtained in experiments performed in the domestic cat. Conception rates after AI in wild and domestic cats are also reported.     

A multiplex PCR for the detection of Brucella spp. and Leptospira spp. DNA from aborted bovine fetuses

Bovine brucellosis and leptospirosis are important causes of bovine abortion around the world. Both diseases can be serologically diagnosed, but many factors may cause false positive and negative results. Direct methods based on bacteriological isolation are usually employed, but they are difficult, time consuming and dangerous. Monoplex polymerase chain reaction (PCR) have been successfully described for the detection of Brucella spp. and Leptospira spp. Aiming at improvement in the direct diagnosis, a multiplex PCR (mPCR) for the detection of these agents in aborted bovine fetuses is described. The detection threshold of the mPCR was evaluated in experimentally contaminated bovine clinical samples using a conventional proteinase K/SDS or a boiling-based extraction protocols. The mPCR was applied to two groups of clinical samples: 63 episodes of bovine abortion and eight hamsters experimentally infected with Leptospira interrogans serovar pomona. Adopting microbiological isolation as reference, the test showed a sensitivity of 100% in both groups of clinical samples. Seven samples collected from bovine fetuses were Brucella spp. culture negative but showed positive results in mPCR. Regarding Leptospira spp. detection, similar results were observed in three bovine clinical samples. All hamsters infected with Leptospira were positive in both microbiological culture and mPCR. The boiling extraction protocol showed better results in some clinical samples, probably by the removal of PCR inhibitors by heat treatment. The high sensitivity, simplicity and the possibility of detection of both bacteria in a single tube reaction support the use of the mPCR described in the routine diagnosis.     

Deletion of the GI-2 integrase and the wbkA flanking transposase improves the stability of Brucella melitensis Rev 1 vaccine

Brucella melitensis Rev 1 is the best vaccine available for the prophylaxis of small ruminant brucellosis and, indirectly, for reducing human brucellosis. However, Rev 1 shows anomalously high rates of spontaneous dissociation from smooth (S) to rough (R) bacteria, the latter being inefficacious as vaccines. This S-R instability results from the loss of the O-polysaccharide. To overcome this problem, we investigated whether some recently described mechanisms promoting mutations in O-polysaccharide genes were involved in Rev 1 S-R dissociation. We found that a proportion of Rev 1 R mutants result from genome rearrangements affecting the wbo O-polysaccharide loci of genomic island GI-2 and the wbkA O-polysaccharide glycosyltransferase gene of the wbk region. Accordingly, we mutated the GI-2 int gene and the wbk IS transposase involved in those arrangements, and found that these Rev 1 mutants maintained the S phenotype and showed lower dissociation levels. Combining these two mutations resulted in a strain (Rev 2) displaying a 95% decrease in dissociation with respect to parental Rev 1 under conditions promoting dissociation. Rev 2 did not differ from Rev 1 in the characteristics used in Rev 1 typing (growth rate, colonial size, reactivity with O-polysaccharide antibodies, phage, dye and antibiotic susceptibility). Moreover, Rev 2 and Rev 1 showed similar attenuation and afforded similar protection in the mouse model of brucellosis vaccines. We conclude that mutations targeting genes and DNA sequences involved in spontaneous O-polysaccharide loss enhance the stability of a critical vaccine phenotype and complement the empirical stabilization precautions taken during S Brucella vaccine production.     

Aeration strategy for controlling grain storage based on simulation and on real data acquisition

In this work a new control strategy for grain aeration systems was proposed. This strategy was implemented in a software package called AERO and relates four conditions which depend on real time data acquisition and on the process simulation. The main advantage of AERO controller is that it can be used in different geographic regions, during different seasons and with different aeration systems, automatically adjusting its set points. The objectives of AERO controller are to equalize the temperatures inside the bin, to cool the grain mass whenever is possible and to maintain the moisture content under safe conditions. The results showed that the proposed strategy is efficient to achieve these objectives. Moreover, significant energy saving has been achieved with the AERO controller.     

A structural mechanism for MscS gating in lipid bilayers

The mechanosensitive channel of small conductance (MscS) is a key determinant in the prokaryotic response to osmotic challenges. We determined the structural rearrangements associated with MscS activation in membranes, using functorial measurements, electron paramagnetic resonance spectroscopy, and computational analyses. MscS was trapped in its open conformation after the transbilayer pressure profile was modified through the asymmetric incorporation of lysophospholipids. The transition from the closed to the open state is accompanied by the downward tilting of the transmembrane TM1-TM2 hairpin and by the expansion, tilt, and rotation of the TM3 helices. These movements expand the permeation pathway, leading to an increase in accessibility to water around TM3. Our open MscS model is compatible with single-channel conductance measurements and supports the notion that helix tilting is associated with efficient pore widening in mechanosensitive channels.     

Comment on "Drought-induced reduction in global terrestrial net primary production from 2000 through 2009"

Zhao and Running (Reports, 20 August 2010, p. 940) reported a reduction in global terrestrial net primary production (NPP) from 2000 through 2009. We argue that the small trends, regional patterns, and interannual variations that they describe are artifacts of their NPP model. Satellite observations of vegetation activity show no statistically significant changes in more than 85% of the vegetated lands south of 70°N during the same 2000 to 2009 period.     

Effects of no‐take area size and age of marine protected areas on fisheries yields: a meta‐analytical approach

Marine protected areas (MPAs) are often promoted as tools for biodiversity conservation as well as for fisheries management. Despite increasing evidence of their usefulness, questions remain regarding the optimal design of MPAs, in particular concerning their function as fisheries management tools, for which empirical studies are still lacking. Using 28 data sets from seven MPAs in Southern Europe, we developed a meta‐analytical approach to investigate the effects of protection on adjacent fisheries and asking how these effects are influenced by MPA size and age. Southern European MPAs showed clear effects on the surrounding fisheries, on the ‘catch per unit effort’ (CPUE) of target species, but especially on the CPUE of the marketable catch. These effects depended on the time of protection and on the size of the no‐take area. CPUE of both target species and the marketable catch increased gradually by 2–4% per year over a long time period (at least 30 years). The influence of the size of the no‐take area appeared to be more complex. The catch rates of the entire fishery in and around the MPA were higher when the no‐take areas were smaller. Conversely, catch rates of selected fisheries that were expected to benefit most from protection increased when the no‐take area was larger. Our results emphasize the importance of MPA size on its export functions and suggest that an adequate, often extended, time frame be used for the management and the evaluation of effectiveness of MPAs.     

Antiparasitic efficacy of dietary administration of trichlorfon (Masoten®) in the control of Neoechinorhynchus buttnerae (Neochinorhynchidae) in Colossoma macropomum (Serrasalmidae)

Aquaculture International - This study investigated the efficacy of trichlorfon against Neoechinorhynchus buttnerae, as well as its palatability, residue analysis, and the blood parameters of...