Is sperm cryopreservation in absence of permeable cryoprotectants suitable for subfertile donkeys?

Sperm from fertile donkeys have been successfully frozen in absence of permeable cryoprotectants. The aim of this study was to determine whether this cryopreservation method is suitable for subfertile donkeys in comparison to conventional sperm freezing with glycerol. Ejaculates were collected from four Andalusian Donkeys: three fertile and one subfertile. Semen was frozen with an extender containing glycerol (GLY), or adding instead sucrose 0.25 molar and 1% bovine serum albumin (SUC) as non‐permeable cryoprotectants. After thawing, samples were assessed for total (TM, %) and progressive (PM, %) sperm motility by CASA, plasma membrane integrity (PMI, %) by epifluorescence microscopy and DNA integrity (DFI, %) by SCSA. Results (mean ± SD) were compared between extenders in fertile and subfertile donkeys using the Student's t test. No differences between GLY and SUC treatments were found in the fertile group for the sperm parameters assessed. In subfertile donkey ejaculates, GLY resulted in significantly higher values than SUC for TM (25.5 ± 3.1 vs. 19.6 ± 1.9) and PM (13.3 ± 5.1 vs. 4.0 ± 1.2), respectively. In conclusion, considering all the sperm parameters assessed, sperm freezing in absence of permeable cryoprotectants may not be still an option for cryopreservation of subfertile donkey sperm.     

Linkage disequilibrium and effective population size in Gir cattle selected for yearling weight

Linkage disequilibrium (LD) plays an important role in genomic selection and mapping of quantitative trait loci (QTL). This study investigated the pattern of LD and effective population size (N_e) in Gir cattle selected for yearling weight. For this purpose, 173 animals with imputed genotypes (from 18 animals genotyped with the Illumina BovineHD BeadChip and 155 animals genotyped with the Bovine LDv4 panel) were analysed. The LD was evaluated at distances of 25–50 kb, 50–100 kb, 100–500 kb and 0.5–1 Mb. The N_e was estimated based on 5 past generations. The r² values (a measure of LD) were, respectively, .35, .29, .18 and .032 for the distances evaluated. The LD estimates decreased with increasing distance of SNP pairs and LD persisted up to a distance of 100 kb (r² = .29). The N_e was greater in generations 4 and 5 (24 and 30 animals, respectively) and declined drastically after the last generation (12 animals). The results showed high levels of LD and low N_e, which were probably due to the loss of genetic variability as a consequence of the structure of the Gir population studied.     

Antibody response to an anti-rabies vaccine in a dog population under field conditions in Bolivia

Rabies remains an important public health issue in Bolivia, South America. Public concern and fears are most focussed on dogs as the source of rabies. The objective of the present study was to assess immunity of an inactivated suckling mouse brain vaccine against canine rabies used for the official vaccination campaigns under field conditions in an endemic area of rabies in Bolivia. A total of 236 vaccinated and 44 unvaccinated dogs in Santa Cruz de la Sierra, selected using stratified random sampling, were investigated in order to obtain owned dog characteristics and antibody titres against rabies in April 2007. The proportion of vaccinated dogs with an antibody titre exceeded the protection threshold value of 0.5 EU/ml was 58% [95% confidence intervals (CI): 52-65], indicating that vaccination is likely to elicit an antibody response (odds ratio 6.3, 95% CI: 1.2-11.5). The range of geometric mean of antibody titre for vaccinated dogs (0.89 EU/ml; 95% CI: 0.75-1.04) was considered to meet the minimal acceptable level indicating an adequate immune response to the vaccine. However, the titre level was not satisfactory in comparison with the results from other field investigations with inactivated tissue culture vaccines. It is recommended for public health authorities to (1) consider modernizing their vaccine manufacturing method because the level of immunity induced by the current vaccine is comparably low, (2) conduct frequent vaccination campaigns to maintain high levels of vaccination coverage, and (3) actively manage the domestic dog population in the study area, which is largely responsible for rabies maintenance.     

Retrospective analysis of selected predictors of mortality within a veterinary intensive care unit

The records of 204 cats entering the intensive care unit (ICU) at the University of Edinburgh Small Animal Hospital between December 2002 and October 2006 were retrospectively analysed. Of these, 37 cats over 12 months of age had a systolic blood pressure recorded on entry into the ICU, and this group comprised our study population. Of these 37 cats, 36 had both heart rate and respiratory rate recorded on entry into the ICU, whilst 24 of these cats also had body temperature recorded. The relationship between (i) survival to discharge and (ii) survival until 21 days after admission to the ICU was analysed using univariate generalised linear models with binomial errors. The robustness of any significant relationship was assessed using multivariate analysis methods. In addition, receiver operator curves (ROC) were generated for any of the significant predictors of mortality and from these curves the threshold values, optimal sensitivity and specificity were calculated. Using these values survival curves were generated for any significant prognostic indexes. A decreased blood pressure at the time of admission to the ICU was found to be a significant negative predictor of survival until discharge from the hospital. Overall, a systolic blood pressure of 124 mmHg or higher at the time of admission to the ICU has a sensitivity of 47.8% and a specificity of 85.7% for predicting that a cat will survive until discharge from the hospital.     

Environmental impact of razor clam harvesting using salt in Ria Formosa lagoon (Southern Portugal) and subsequent recovery of associated benthic communities

• 1. Razor clams are found in different habitats ranging from sheltered systems (e.g. lagoons and estuaries) to open coasts. They are distributed worldwide and comprise a small number of species of high economic value. Depending on the specific habitat and species exploited, different mechanical and hand‐harvesting techniques are employed. While the environmental effects resulting from mechanized methods have been described by several authors, the impacts caused by traditional hand‐harvesting methods remain unknown. Therefore, a study was undertaken in Ria Formosa lagoon (South of Portugal) addressing the environmental effects resulting from harvesting Solen marginatus with salt.
• 2. No significant impact on the sediment was found; the main effect was an increase in salinity after covering the area with salt, which decreased rapidly with the flood tide and after a few hours had returned to pre‐harvesting levels.
• 3. No effects on benthic communities were observed, with similar fluctuation patterns recorded in control and experimental areas, the observed differences being attributed to the natural variability of benthic populations.
• 4. Based on the results obtained in the present study, a razor clam fishery using salt in intertidal areas can be considered environmentally ‘friendly’.

Copyright © 2008 John Wiley & Sons, Ltd.     

Activity pattern of the marine shrimp Litopenaeus vannamei (Boone 1931) in laboratory as a function of different feeding frequencies

Management practices on most Brazilian shrimp farms have resulted in high expenses for producers and damage to the environment. Applied ethology could provide information on the pattern of shrimp activity, enabling more efficient feeding frequencies. Behavioural activities of the shrimp Litopenaeus vannamei were recorded during different feeding frequencies. The shrimp were kept in aquariums on a 12:12 h light/dark cycle. They were fed commercial ration three (at 6:00, 12:00 and 18:00 hours), four (at 6:00, 10:00, 14:00 and 18:00 hours) or seven times per day (at 6:00. 8:00, 10:00, 12:00, 14:00, 16:00 and 18:00 hours). We observed animals at 15 min^?1 aquarium^?1 time periods, recording feeding, substrate exploration, swimming and inactivity. Feeding and exploration were the highest for shrimp fed three times per day, swimming was greatest when animals received four feedings, whereas inactivity was higher for shrimp fed seven times per day. There was greater food ingestion between 12:00 and 14:00 hours for animals fed three and four times per day, whereas swimming was high mainly at 18:00 hours for shrimp fed three and seven times per day. The results indicate higher forage‐related activities (exploration/ingestion) when feed was offered three or four times, suggesting optimization in the pattern of shrimp activities based on these feeding frequencies.     

From Natural Xanthones to Synthetic C-1 Aminated 3,4-Dioxygenated Xanthones as Optimized Antifouling Agents

Biofouling, which occurs when certain marine species attach and accumulate in artificial submerged structures, represents a serious economic and environmental issue worldwide. The discovery of new non-toxic and eco-friendly antifouling systems to control or prevent biofouling is, therefore, a practical and urgent need. In this work, the antifouling activity of a series of 24 xanthones, with chemical similarities to natural products, was exploited. Nine (1, 2, 4, 6, 8, 16, 19, 21, and 23) of the tested xanthones presented highly significant anti-settlement responses at 50 μM against the settlement of mussel Mytilus galloprovincialis larvae and low toxicity to this macrofouling species. Xanthones 21 and 23 emerged as the most effective larval settlement inhibitors (EC₅₀ = 7.28 and 3.57 µM, respectively). Additionally, xanthone 23 exhibited a therapeutic ratio (LC₅₀/EC₅₀) > 15, as required by the US Navy program attesting its suitability as natural antifouling agents. From the nine tested xanthones, none of the compounds were found to significantly inhibit the growth of the marine biofilm-forming bacterial strains tested. Xanthones 4, 6, 8, 16, 19, 21, and 23 were found to be non-toxic to the marine non-target species Artemia salina (<10% mortality at 50 μM). Insights on the antifouling mode of action of the hit xanthones 21 and 23 suggest that these two compounds affected similar molecular targets and cellular processes in mussel larvae, including that related to mussel adhesion capacity. This work exposes for the first time the relevance of C-1 aminated xanthones with a 3,4-dioxygenated pattern of substitution as new non-toxic products to prevent marine biofouling.     

Dietary organic acids blend alone or in combination with an essential oil on the survival,growth, gut/liver structure and de hemato‐immunological in Nile tilapia Oreochromis niloticus

This study evaluated the effects of dietary supplementation of organic acids blend (OAB) alone or in combination with essential oil, Lippia origanoides (OAE) for Nile tilapia fed supplemented diets for 30 days. Fish (1.1 ± 0.04 g) were fed control (Control), or OAB 0.5% or OAB 0.5% + essential oil 0.125% (OAE) respectively. At the end of the experiment, samples were collected for de hemato‐immunological, histological analysis of the intestine and liver, as well as microbiology of the intestine. The pH of the diets supplemented with OAB and OAE reduced 0.92 and 0.19 respectively. The growth and FCR were unaffected by the treatments, but survival was significantly higher in the OAB treatment. Fish fed the OAB diet showed reduced concentration of total heterotrophic bacteria and Pseudomonas sp. in the intestine. Increased glucose in fish fed OAB and high number of circulating monocytes in fish fed OAE diet were observed. The anterior intestine of fish fed OAE diet showed larger number of goblet cells and increased villi height. The diet supplemented with OAB, mainly, improved the intestinal health and survival of tilapia juveniles and can be used in juvenile production.     

Evaluation of the Microalgae Paste Viability Produced in a Mollusk Hatchery in Southern Brazil

The present study was conducted to define a methodology to produce and store small‐scale microalgae paste to be used in a mollusk hatchery. Microalgae were cultured in 500 L fiberglass tanks, under temperature of 20 ± 2 C, Guillard f/2 culture medium, and continuous light intensity of 203–226 μmol photons/m²/sec. Cultures were centrifuged at 2000 g at the exponential growth phase. Microalgae cell quality after centrifugation and during storage was determined by analyses with Evan’s blue stain and by counting the number of total marine bacteria. Treatments with and without additive were applied to the microalgae paste produced, which was distributed into 100 mL plastic containers, capped, and stored under refrigeration at 4 ± 1 C. Results indicated that in the Chaetoceros muelleri paste, centrifugation did not damage the cells and the number of total marine bacteria reduced significantly from 2.9 × 10⁶ to 8.3 × 10⁵ colony‐forming units per milliliter. Chaetoceros muelleri and Chaetoceros calcitrans pastes stored with addition of 0.1% ascorbic acid had a shelf life shorter than 2 wk. For the treatment without additive, results with Evan’s blue stain showed that cells (99%) remained viable until the sixth week of storage for C. muelleri and seventh week of storage for Skeletonema sp. and C. calcitrans. The number of bacteria did not increase during storage for C. calcitrans and Skeletonema (P > 0.05). For C. muelleri, an increase in bacteria (P < 0.05) was observed after the sixth week of storage. This study demonstrated the feasibility to produce and store microalgae paste for a period of 2–8 wk, which allows it to be used as food source and also optimizes the use of microalgae cultured in laboratory.