Use of Powdered Egg Yolk vs Fresh Egg Yolk for the Cryopreservation of Ovine Semen

Egg yolk is a common additive to sperm cryopreservation diluents. Because of its animal origin, however, it also represents a potential risk of microbiological contamination in the diluent. This potential contamination can be avoided by using powdered egg yolk, instead of fresh egg yolk, as it is pasteurized. This study was conducted to determine ram sperm cryosurvival was affected by the type of egg yolk used (powdered egg yolk or fresh egg yolk) and by yolk concentration (10, 15 or 20%) in the diluent. Microbiological analyses were also performed to quantify the microbiological contamination in the diluents containing the two types of egg yolk. Sperm cryosurvival was determined by motility and morphology analyses after thawing. Motility parameters were assessed using a computer-assisted sperm analysis (CASA) system, and the percentage of sperm with a normal apical ridge was evaluated using a differential interference contrast microscope. No significant differences were observed between diluents in the percentage of sperm with normal apical ridge. However, higher percentages of total motile cells were observed for samples containing powdered egg yolk (69%) compared to samples containing fresh egg yolk (60%). However, sperm in diluents containing fresh egg yolk, exhibited higher values for average-path velocity, straight-line velocity and beat cross frequency and lower values for amplitude of lateral head displacement (p <0.05), compared to cells in diluents containing powdered egg yolk. Microbiological contamination was similar (<200 CFU/ml) in both diluents, and no bacterial growth was observed in either, when antibiotics were added. Therefore, powdered egg yolk can be effective used in diluents for the freezing of ram semen. However, the in vivo fertility of sperm frozen in diluents containing powdered egg yolk should be tested, as some motility parameters were different for sperm treated with powdered egg yolk compared to fresh egg yolk.     

Occupancy pattern of a long-horned beetle in a variegated forest landscape: linkages between tree quality and forest cover across spatial scales

Context

Interactions between landscape-scale processes and fine-grained habitat heterogeneity are usually invoked to explain species occupancy in fragmented landscapes. In variegated landscapes, however, organisms face continuous variation in micro-habitat features, which makes necessary to consider ecologically meaningful estimates of habitat quality at different spatial scales.

Objectives

We evaluated the spatial scales at which forest cover and tree quality make the greatest contribution to the occupancy of the long-horned beetle Microplophorus magellanicus (Coleoptera: Cerambycidae) in a variegated forest landscape.

Methods

We used averaged data of tree quality (as derived from remote sensing estimates of the decay stage of single trees) and spatially independent pheromone-baited traps to model the occurrence probability as a function of multiple cross-scale combinations between forest cover and tree quality (with scales ranging between 50 and 400 m).

Results

Model support and performance increased monotonically with the increasing scale at which tree quality was measured. Forest cover was not significant, and did not exhibit scale-specific effects on the occurrence probability of M. magellanicus. The interactive effect between tree quality and forest cover was stronger than the independent (additive) effects of tree quality and particularly forest cover. Significant interactions included tree quality measured at spatial scales ≥200 m, but cross-scale interactions occurred only in four of the seven best-supported models.

Conclusions

M. magellanicus respond to the high-quality trees available in the landscape rather than to the amount of forest per se. Conservation of viable metapopulations of M. magellanicus should consider the quality of trees at spatial scales >200 m.

     

Cultivation of Pen Shells: An Example with Atrina maura in Northwestern Mexico

Pen shells are economically important and most are threatened by overexploitation and habitat degradation. Cultivation represents an alternative for preserving the resource and increasing the production. Atrina maura was cultivated in a suspension system which was followed by a bottom phase. Samples of pen shells and tissues were collected monthly to measure absolute growth, shell height increase, and an adductor muscle index. After the trials, adults shell height was 195 ± 10 mm, total weight was 223 ± 28 g, and mean shell height increase was 0.81 cm/mo. Protective devices were vital to avoid predation by crabs and fish; survival was 90% in the suspension phase and 70% in the bottom phase. Variations in the muscle index were apparently related to reproduction, suggesting activity (partial spawning) from August 2008 to April 2009 in juveniles and a major spawning in adults during August 2009. Adductor muscles reached a mean of 21 g after 20 mo of cultivation, but muscle weights decreased significantly after reproduction (August to November 2009). Results indicated that A. maura is suitable for aquaculture and the cultivation system adopted here should be tested in pilot‐scale ventures. This system may work with other species of pen shells.     

Endometrial Tissue Concentrations of Enrofloxacin after Intrauterine Administration to Mares

Endometritis in mares is a common cause of infertility. Conventional treatments of the disease have mostly been unsuccessful, so new therapeutic alternatives need to be investigated. This study evaluated the uterine disposition and plasma pharmacokinetic behaviour of a commercial formulation of enrofloxacin (EFX) given by the intrauterine (i.u.) route (2.5 mg/kg) in healthy mares. In order to evaluate the uterine inflammatory response, an initial histopathological study assessing polymorphonuclear cell infiltration was carried out in 20 mares over a 14-day period after treatment. In a second study, 6 healthy adult mares were used for the pharmacokinetic study. Samples of uterine tissue and plasma were collected from 0 to 24 h after the i.u. treatment with 5% EFX solution. Samples were analysed by conventional microbiological assay using an EFX-sensitive strain of Escherichia coli (ATCC 25922). There was a moderate but statistically nonsignificant inflammatory response following i.u. administration of either the formulation or the vehicle alone. Pharmacokinetic analysis of the uterine concentrations of EFX showed a slow and sustained depletion, with EFX remaining at concentrations above the MIC for 24 h after treatment. The area under the concentration–time curve obtained for the uterus suggested that EFX and its metabolites are specifically retained in the uterus, which is the target tissue for bacterial colonization. Neither study provided any evidence of EFX toxicity. In conclusion, these results are encouraging and suggest that EFX may be a useful local treatment in mares with bacterial endometritis.     

Immune and clinical parameters associated with Leishmania infantum infection in the golden hamster model

For experimental infections with viscerotropic strains of Leishmania, a suitable animal model is not yet defined. In the present work, we have reappraised the use of golden hamster (Mesocricetus auratus) as an experimental model for infection with Leishmania infantum. Groups of hamsters were challenged by the intracardial route with doses ranging from 10(3) to 10(5) infectious promastigotes and the animals were monitored for 1-year follow-up period. The outcome of the infection was assessed by clinical symptoms of leishmaniasis, parasite loads in both liver and spleen, humoral response to Leishmania antigens and antibody levels in kidneys. The humoral response was analysed using either crude antigens (by ELISA and Western blotting) or several recombinant Leishmania antigens (Hsp70, Hsp83, LiP2a, LiP2b, H2A, H3 and KMP-11). From the analysis of all these parameters, we established the existence of three groups of animals: symptomatic or susceptible, oligosymptomatic, and resistant. Given the parallelism existing between the outcomes of Leishmania-infection in hamsters, dogs and humans, we believe that our data illustrate that the hamster is an excellent experimental model to study visceral leishmaniasis and for the design of vaccine development.     

Pesticide use and residues on Queensland wool

Objective To determine practices for control of louse infestation and blowfly strike in Queensland sheep flocks that are associated with organophosphorous and synthetic pyrethroid residues on wool.
Design Information on residues was obtained from a survey of Queensland wool clips. Information on pesticide use was obtained from a trace-back postal survey. The association between pesticide use and residues was assessed using generalised linear models, controlling for potential confounding by flock location.
Procedure Between 1995 and 1997 Queensland wool clips were randomly sampled. Samples were tested for the presence and amount (mg per kg of greasy wool) of organophosphorous and synthetic pyrethroid pesticides. A questionnaire seeking information on flock characteristics and pesticide use was sent to the manager of each flock from which a wool sample was tested.
Results The median amount of OP and SP residue was 0.8 and 0.25 mg/kg, respectively, and 91 and 95% of wool samples contained < 8 mg/kg of OP and SP residues, respectively. The frequency of OP pesticide use for louse control was significantly (P = 0.005) associated with mean OP residue amount, and the timing of SP use for louse control, in relation to shearing, was significantly (P < 0.001) associated with mean SP residue amount.
Conclusion Most Queensland wool clips have acceptable amounts of residues after the use of OP and SP pesticides, but wool growers can further reduce residues by effectively controlling louse infestation with pesticide applications early after shearing and the use of non-chemical methods of ectoparasite control.     

Narrow genetic base in forest restoration with holm oak (Quercus ilex L.) in Sicily

In order to empirically assess the effect of actual seed sampling strategy on genetic diversity of holm oak (Quercus ilex) forestations in Sicily, we have analysed the genetic composition of two seedling lots (nursery stock and plantation) and their known natural seed origin stand by means of six nuclear microsatellite loci. Significant reduction in genetic diversity and significant difference in genetic composition of the seedling lots compared to the seed origin stand were detected. The female and the total effective number of parents were quantified by means of maternity assignment of seedlings and temporal changes in allele frequencies. Extremely low effective maternity numbers were estimated (Nf _e ≈ 2–4) and estimates accounting for both seed and pollen donors gave also low values (N _e ≈ 35–50). These values can be explained by an inappropriate forestry seed harvest strategy limited to a small number of spatially close trees.