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.     

Biological and socioeconomic implications of recreational boat fishing for the management of fishery resources in the marine reserve of Cap de Creus (NW Mediterranean)

A survey of coastal recreational boat fishing was conducted in summer 2006 in the marine reserve of Cap de Creus (NW Mediterranean) to assess the biological and socioeconomic implications of this leisure activity. Recreational boat fishers employ four different fishing techniques: bottom fishing rod, fluixa, trolling and surface fishing rod. Although the targeted species depend on the fishing method used, a total of 33 fish species were identified in the catch (8 were pelagic or benthopelagic and the rest were demersal). Fishing effort was high since fishers fished an average of 4 h/day, 8 days/month and nearly 6 months/year. Apart from the highly varied exploitation of the fauna, recreational fishing in Cap de Creus has a large economic effect on the local economy since the majority of fishers were visitors who were spending holidays in one of the villages belonging to the Park, where most of expenditures related to angling activities were made. Overall, results highlight the pressure that recreational boat fishing exerts on fish communities, particularly on littoral, demersal ones. Considering these biological and socioeconomic implications, the competition between recreational and artisanal fishers for littoral resources and the low level of compliance with the current sport fishing regulations, the implementation of a comprehensive management strategy in Mediterranean costal areas is needed.     

SDS-PAGE and Western blot of urinary proteins in dogs with leishmaniasis

Canine leishmaniasis is an endemic disease in the Mediterranean area caused by the protozoan Leishmania infantum, which usually produces renal failure. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and Western blot using antibodies to IgG and IgA from dogs were carried out in the urine of 22 dogs with leishmaniasis diagnosed by ELISA and confirmed by PCR, and 20 healthy dogs. The results were compared to renal function laboratory tests and to those from a histopathological study of the kidneys from sick animals that died naturally or were euthanized. Five different bands with molecular weights ranging from 10 to 110 kDa were obtained from the electrophoresis of the urine of healthy dogs. 33.5% of total proteins corresponded to low molecular weight proteins and the other proteins had middle and high molecular weights. However, in the group with leishmaniasis, a maximum of 11 different bands with molecular weights ranging from 10 kDa to 150 kDa were displayed in the electrophoresis of the urine. The urine electrophoretic pattern in the sick dogs was classified as mixed (proteins with high and low molecular weights) because low molecular weight proteins made up 57.9% and the rest of the proteins had middle and high molecular weights. In Western blot, none of the healthy dogs showed excretion of IgG and/or IgA, whereas IgG and IgA were detected in the Western blot of urine of 68% and 55% respectively of dogs with leishmaniasis. The results obtained in the leishmaniasis group agreed with glomerular and tubular damage, which were confirmed by the histopathological findings.     

Serum α-globulin fraction in horses is related to changes in the acute phase proteins

Serum electrophoresis is a useful technique to diagnose some diseases characterized by quantitative and qualitative changes in the serum proteins, such as the acute phase proteins. An increased synthesis of acute phase proteins has been associated with acute infection and inflammation. We wanted to study whether gastrointestinal diseases and disorders of the locomotor system in horses produce enough inflammatory reaction so the acute phase proteins can be detected by electrophoresis in serum. Serum total proteins were calculated, and the serum electrophoresis from 63 horses was performed. The horses were classified in 3 groups: Group I (19 healthy horses), Group II (20 horses suffering from inflammatory disorders of the gastrointestinal tract), and Group III (24 horses with inflammatory disorders of the locomotor system). Results indicated an increase of the acute phase proteins only in patent inflammatory disorders that showed a rapid progression. A global increase in those proteins was observed only for α₁-globulins from Group II.     

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.     

The Control of Reactive Oxygen Species Influences Porcine Oocyte In Vitro Maturation

The aim of this study was to examine the effect of varying intracellular reactive oxygen species (ROS) levels during oocyte in vitro maturation with enzymatic ROS production systems (xanthine + xanthine oxidase or xanthine + xanthine oxidase + catalase), scavenger systems (catalase or superoxide dismutase + catalase) or cysteine on porcine oocyte maturation. Oocyte ROS levels showed an increase when H₂O₂ or O₂?^‐ production systems were added to the culture medium (p < 0.05). On the other hand, the presence of ROS scavengers in the maturation medium did not modify oocyte ROS levels compared with the control after 48 h of maturation, but the addition of cysteine induced a decrease in oocyte ROS levels (p < 0.05). The ROS production systems used in this work did not modified the percentage of oocyte nuclear maturation, but increased the decondensation of sperm head (p < 0.05) and decreased the pronuclear formation (p < 0.05). In turn, the addition of O₂?^‐ and H₂O₂ scavenging systems during in vitro maturation did not modify the percentage of oocytes reaching metaphase II nor the oocytes with decondensed sperm head or pronuclei after fertilization. However, both parameters increased in the presence of cysteine (p < 0.05). The exogenous generation of O₂?^‐ and H₂O₂ during oocyte in vitro maturation would not affect nuclear maturation or later sperm penetration, but most of the spermatozoa cannot progress to form the pronuclei after fusion with the oocyte. The decrease in endogenous ROS levels by the addition of cysteine would improve pronuclear formation after sperm penetration.