Interactions between different media and follicle‐stimulating hormone supplementation on in vitro culture of preantral follicles enclosed in ovarian tissue derived from collared peccaries (Pecari tajacu Linneaus, 1758)

The aim was to verify the effect of follicle‐stimulating hormone (FSH) supplementation to α‐MEM+ or TCM199+ media on the in vitro development of ovarian preantral follicles (PFs) derived from collared peccaries. Ovaries (n = 5 pairs) were collected and divided into fragments destined to control group (non‐cultured) or treatments that were cultured for 7 days. The PFs morphology, growth and activation were evaluated by classical histology. The immunohistochemistry markers Ag‐NOR and PCNA were used for nuclear proliferation analysis, and the picrosirius red labelling was used for ovarian extracellular matrix (ECM) evaluation. After 7‐day culture, only the TCM199+ treatment maintained the proportion of intact PFs similar to day 1 (63.2%), but no differences were found among treatments (p > .05). In addition, a significant increase in the growing follicles proportion was verified for all the treatments, indicating follicular activation (p > .05). By the Ag‐NOR analysis, only the TCM199+/FSH maintained the nuclear proliferation similar to the first day (p > .05). The picrosirius red staining revealed that the ECM remained intact in all the treatments (p > .05). We suggest the use of TCM199+ medium supplemented of FSH for the in vitro development of peccaries PFs under 7‐day culturing conditions.     

Validation of the periodicity of growth zone formation in the otoliths of four fish species from the Upper Zambezi ecoregion,southern Africa

In the Upper Zambezi and Okavango ecoregions, Brycinus lateralis, Hepsetus cuvieri, Schilbe intermedius and Serranochromis macrocephalus are important in subsistence fisheries, while S. intermedius and S. macrocephalus are often caught in commercial catches. Despite their importance, there is little information on their age and growth and age validation. Growth zone deposition rate in otoliths of B. lateralis, H. cuvieri, S. intermedius and S. macrocephalus was validated as annual using edge analysis in this study. Annual deposition of growth zones was corroborated by a fluorochrome marking experiment. Both methods demonstrated that one growth zone was deposited annually. Edge analysis demonstrated that the highest proportion of opaque zones was detected between August and December, most likely as a result of slow growth during lower temperature water during the preceding winter. With growth zone deposition validated as annual, future research should focus on determining the growth, maturity and mortality rates of these species, parameters which are important for their management.     

Implementation of two high through-put techniques in a novel application: detecting point mutations in large EMS mutated plant populations

Background  

The establishment of mutant populations together with the strategies for targeted mutation detection has been applied successfully to a large number of organisms including many species in the plant kingdom. Considerable efforts have been invested into research on tomato as a model for berry-fruit plants. With the progress of the tomato sequencing project, reverse genetics becomes an obvious and achievable goal.     

In-field calibration of a dielectric soil moisture meter designed for use in an access tube

Abstract. The in-field calibration of a dielectric probe to measure soil water content is described. The probe uses an access tube analogous to that of the neutron probe. The dielectric constant was measured at soil depths of 10, 20, 30, 40, 60 and 100 cm. Cores of soil were then taken from the face of pits dug 30 cm from the access tube and their soil water contents determined by oven drying. The dielectric constant values measured by the probe were calibrated against water contents from these cores. We found that sensor depth needed to be included to achieve a good calibration model that explained 72% of the variance. It is argued that depth needs to be included because of artefacts introduced during the installation of the access tube.     

Enhanced efficiency of electrostatically charged insecticide aerosols

The bioefficacy of a standard domestic aerosol insecticide has been compared with that of a similar spray in which the droplets were electrostatically charged. The aerosol was charged without the need for an external power supply, by enhancing natural charge separation processes that occur during atomisation. The charge-to-mass ratio achieved was 1.1 x 10(-4) C kg-1, compared with 2.2 x 10(-5) C kg-1 for the standard aerosol. Efficiency was assessed for insects in free flight. A direct space-spray application method was used to study the bioefficacy of these aerosols on houseflies, Musca domestica, and an indirect space-spray method on houseflies and mosquitoes, Culex quinquefasciatus. Two levels of concentration of active ingredients were compared. At low concentrations of 1.57 g kg-1 of bioallethrin and 0.29 g kg-1 of bioresmethrin, the charged aerosol achieved a significant reduction in KDT50 (time at which 50% of the flies were knocked down), compared with the standard aerosol. In the direct spray application to houseflies, the KDT50 was reduced by 50%, while in the indirect spray the KDT50 was reduced by 40%. In the indirect spray of mosquitoes, the KDT50 was reduced by 22%. With higher active ingredient concentrations of 2.09 g kg-1 for bioallethrin and 0.39 g kg-1 for bioresmethrin, the charged aerosol also demonstrated faster knockdown than the standard, but the improvement was less marked. In the direct spray application to houseflies, the KDT50 was reduced by 21% compared with the standard aerosol, while in the indirect spray the KDT50 was reduced by 16%. In the indirect spray of mosquitoes, the KDT50 was reduced by 24%. The percentage mortality after 24 h was also increased with the charged aerosol. Charge carried by the droplets results in increased interception of the insecticide, as the droplets are attracted to the insects in flight and space charge effects within the aerosol could cause improved dispersion in the test chamber.     

Current distribution of sarcoptic mange in wombats

Objective To determine the distribution and prevalence of sarcoptic mange in wombats, particularly the common wombat ( Vombatus ursinus ).
Design Questionnaire survey in two parts.
Procedure Questionnaires were distributed to biologists, rangers, animal carers and naturalists. Part 1 of the questionnaire aimed to determine the present distribution of sarcoptic mange in wombats (103 responses). Part 2 invited respondents to assess the prevalence of sarcoptic mange in wombats over a 3 month period (four responses). Information on wombats from 66 localities was received. Each locality represented an area of about 2500 km².
Results Mange was observed at 93% of localities surveyed and Sarcoptes scabiei was present in common wombats at 52% of localities. Sarcoptic mange was highly prevalent (22%) in two common wombat populations in Victoria. Anecdotal evidence suggested that mange epizootics are sporadic, cause significant morbidity and mortality and have a substantial effect on local abundance. The respondents did not report sarcoptic mange in either northern hairy-nosed wombats ( Lasiorhinus krefftii ) or southern hairy-nosed wombats ( Lasiorhinus latifrons ).
Conclusions Sarcoptic mange occurs in common wombat populations throughout the range of the common wombat including Tasmania and Flinders Island. While mange epizootics are sporadic, they have the potential to threaten the long-term survival of small, remnant populations.     

Effect of Oil Overlay on Inhibition Potential of Roscovitine in Sheep Cumulus‐Oocyte Complexes

Inhibitors of cyclin‐dependent kinases, as roscovitine, have been used to prevent the spontaneous resumption of meiosis in vitro and to improve the oocyte developmental competence. In this study, the interference of oil overlay on the reversible arrest capacity of roscovitine in sheep oocytes as well as its effects on cumulus expansion was evaluated. For this, cumulus‐oocyte complexes (COCs) were cultured for 20 h in TCM 199 with 10% foetal bovine serum (Control) containing 75 μm roscovitine (Rosco). Subsequently, they were in vitro matured (IVM) for further 18 h in inhibitor‐free medium with LH and FSH. The culture was performed in Petri dishes under mineral oil (+) or in 96 well plates without oil overlay (?) at 38.5°C and 5% CO₂. At 20 and 38 h, the cumulus expansion and nuclear maturation were evaluated under stereomicroscope and by Hoechst 33342 staining, respectively. No group presented cumulus expansion at 20 h. After additional culture with gonadotrophins, a significant rate of COCs from both Control groups (+/?) exhibited total expansion while in both Rosco groups (+/?) the partial expansion prevailed. Among the oocytes treated with roscovitine, 65.2% were kept at GV in the absence of oil overlay while 40.6% of them reached MII under oil cover (p < 0.05). This meiotic arrest was reversible, and proper meiosis progression also occurred in the Control groups (+/?). So, the culture system without oil overlay improved the meiotic inhibition promoted by roscovitine without affecting the cumulus expansion rate or the subsequent meiosis progression.     

Assessment of Suitable Porcine Semen for Freezing,According to the Ejaculate Characteristics in the Iberico × Landrace Breed

A total of 35 ejaculates were studied in order to assess the suitability of porcine semen for freezing according to the ejaculate characteristics. The effects of the freezing procedure were identified; a decrease in motility and acrosome quality was found after thawing. The best results on motility were linked to the ejaculates with a volume of less than 100 ml of the sperm‐rich fraction, a concentration lower than 450 × 10⁶ spermatozoa/ml and an agglutination score below 2. However, the best normal apical ridge (NAR) was found when the volume of the sperm‐rich fraction was greater than 150 ml. For this reason, an intermediate volume of the sperm‐rich fraction of the ejaculate for the best motility and the best NAR, a concentration lower than 450 × 10⁶ spermatozoa/ml and a rate of agglutination below 2 should provide the best quality after freezing. This study also attempted to determine whether a positive effect of ejaculate selection on the overall freezing performance might be expected.     

Adverse effects of simulated harvesting of short-rotation willow and poplar coppice on vertical pressures and rut depths

Little is known of the effects of mechanized harvesting on ground conditions during the harvesting of short-rotation coppice. An investigation was therefore carried out in which different vehicles were used to simulate the effects of wheeling from heavy and light harvesters and crop removal equipment. The experiments were carried out on sites containing Bowles hybrid willow (Salix viminalis) and poplars (Populas rap) and on clay and sandy loam soils. The effect of different vehicles was assessed in terms of rut damage and direct measurements of soil stress using buried sensors. Maximum stresses measured 0.3 m below tractor wheels ranged from 50 to 200 kPa, but the greatest stresses, 350 kPa, were recorded under laden trailer wheels. Maximum stresses measured beneath crawler tracks were only 25 kPa. Similarly, substantial ruts were caused by vehicles simulating wheeled harvesters, the deepest ruts were caused by laden trailers but crawler tracks created least disturbance. Wheeling was carried out at soil water contents above the plastic limit and the deepest ruts were created on clay rather than sandy loam soil. The effects of the stresses generated in the soil could impede future root growth, and the deeper ruts formed could damage existing root systems of coppice.