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.     

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.     

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.     

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.     

In vitro sensitivity of the amphibian pathogen Batrachochytrium dendrobatidis to antifungal therapeutics

Chytridiomycosis, a skin disease caused by Batrachochytrium dendrobatidis, has caused amphibian declines worldwide. Amphibians can be treated by percutaneous application of antimicrobials, but knowledge of in vitro susceptibility is lacking. Using a modified broth microdilution method, we describe the in vitro sensitivity of two Australian isolates of B. dendrobatidis to six antimicrobial agents. Growth inhibition was observed, by measurement of optical density, with all agents. Minimum inhibitory concentrations (µg/ml; isolate 1/2) were – voriconazole 0.016/0.008; itraconazole 0.032/0.016; terbinafine 0.063/0.063; fluconazole 0.31/0.31; chloramphenicol 12.5/12.5; amphotericin B 12.5/6.25. Killing effects on zoospores were assessed by observing motility. Amphotericin B and terbinafine killed zoospores within 5 and 30 min depending on concentration, but other antimicrobials were not effective at the highest concentrations tested (100 µg/ml). This knowledge will help in drug selection and treatment optimization. As terbinafine was potent and has rapid effects, study of its pharmacokinetics, safety and efficacy is recommended.     

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.