Cryopreservation of Piau‐Breed Wild Boar Sperm: Assessment of Cooling Curves and Centrifugation Regimes

This study aimed to assess the effects of different cooling curves and centrifugation regimes used in cryopreservation protocols on the post‐thaw viability of Piau‐breed wild boar (Sus scrofa) sperm using in vitro assessment tests. Two centrifugations (800  g for 10 min and 2400  g for 3 min) and two cooling curves (conventional cooling using nitrogen vapour – freezing 1 and automated cooling using a programmed freezing machine – freezing 2) were tested. Therefore, the treatments were divided into M3 – centrifugation at 2400  g for 3 min and freezing 2; M10 – centrifugation at 800  g for 10 min and freezing 2; R3 – centrifugation at 2400  g for 3 min and freezing 1; and R10 – centrifugation at 800  g for 10 min and freezing 1. No significant differences (p > 0.05) between treatments occurred post‐thawing regarding the total sperm motility means recorded. The mean values of the different treatments were not different from each other regarding the supravital staining (SV), hypo‐osmotic test (HO), sperm–egg binding assay or sperm morphology. This study showed that both the cooling curve and the centrifugation regime affected the quality of post‐thaw sperm, and centrifugation for shorter times and cooling curves using automated cooling are the most suitable for minimizing sperm injury.     

Ruminal and morphometric parameters of the rumen and intestines of sheep fed with increasing levels of spineless cactus (Nopalea cochenillifera Salm-Dyck)

Tropical Animal Health and Production - The objective of this study was to evaluate the ruminal parameters (pH, N-NH3, and microbial protein) and morphometry of the rumen and intestine of sheep fed...     

Morphofunctional description of mucous cells in the gills of the Arapaimidae Arapaima gigas (Cuvier) during its development

The gill structure of the Amazonian fish Arapaima gigas (Cuvier 1829) shows ontogenetic changes during development, particularly due the transition from the aquatic to the obligatory air breathing mode of respiration. However, three main cell types can be found in the gills: mitochondrial rich cells, pavement cells and mucous cells (MCs). The MCs are involved in the secretory pathway. The functions of the secreted molecules include mechanical protection of epithelia, protection against parasites and bacterial infection, and role on ion regulation. In this study, we analysed mucous cell location and mucous cell type, based on pH, during the development of A. gigas. Using samples obtained from the environment, gills were collected and fixed in buffered solution. Histological techniques for the identification of MCs were performed Alcian Blue (AB) and periodic acid‐Schiff (PAS). The results showed the presence of PAS+ and AB+ cells in the whole filament in all examined fish. In animals less than 50 g, few MCs were present, and no differences were observed in AB+ and PAS+ cells. In animals weighing close to 500 g, more PAS+ cells than AB+ cells were observed, and in animals that weighed more than 1,000 g, more AB+ cells than PAS+ cells were observed. These observations may be a result of the ontogenetic changes in the gill epithelia, which can change the osmorespiratory compromise in ion regulation functions as well the glycosaminoglycans secreted by PAS cells, which in large animals can play a role in the protection against parasites and bacterial infection.     

Phage Therapy and Photodynamic Therapy: Low Environmental Impact Approaches to Inactivate Microorganisms in Fish Farming Plants

Owing to the increasing importance of aquaculture to compensate for the progressive worldwide reduction of natural fish and to the fact that several fish farming plants often suffer from heavy financial losses due to the development of infections caused by microbial pathogens, including multidrug resistant bacteria, more environmentally-friendly strategies to control fish infections are urgently needed to make the aquaculture industry more sustainable. The aim of this review is to briefly present the typical fish farming diseases and their threats and discuss the present state of chemotherapy to inactivate microorganisms in fish farming plants as well as to examine the new environmentally friendly approaches to control fish infection namely phage therapy and photodynamic antimicrobial therapy.     

Characterization of intracellular and extracellular saxitoxin levels in both field and cultured Alexandrium spp. samples from Sequim Bay, Washington

Traditionally, harmful algal bloom studies have primarily focused on quantifying toxin levels contained within the phytoplankton cells of interest. In the case of paralytic shellfish poisoning toxins (PSTs), intracellular toxin levels and the effects of dietary consumption of toxic cells by planktivores have been well documented. However, little information is available regarding the levels of extracellular PSTs that may leak or be released into seawater from toxic cells during blooms. In order to fully evaluate the risks of harmful algal bloom toxins in the marine food web, it is necessary to understand all potential routes of exposure. In the present study, extracellular and intracellular PST levels were measured in field seawater samples (collected weekly from June to October 2004–2007) and in Alexandrium spp. culture samples isolated from Sequim Bay, Washington. Measurable levels of intra- and extra-cellular toxins were detected in both field and culture samples via receptor binding assay (RBA) and an enzyme-linked immunosorbent assay (ELISA). Characterization of the PST toxin profile in the Sequim Bay isolates by pre-column oxidation and HPLC-fluorescence detection revealed that gonyautoxin 1 and 4 made up 65 ± 9.7 % of the total PSTs present. Collectively, these data confirm that extracellular PSTs are present during blooms of Alexandrium spp. in the Sequim Bay region.     

Cadmium uptake by Lupinus albus (L.): Cadmium excretion,a possible mechanism of cadmium tolerance

Cadmium (Cd) uptake by white lupin (Lupinus albus) was studied at low Cd concentrations (0.05nM to 5 μM) in hydroponic solution. Ten 12‐day old seedlings were pretreated in 0.5 mM CaCl₂ solution in presence and absence of metabolic inhibitors (DCCD, DNP or NaN3). Cadmium solutions were labelled with carrier free ¹⁰⁹CdCl2. Cadmium uptake was measured after a 2 h desorption in unlabelled CdCl2 solution. In the absence of any metabolic inhibitor and at 5 [μM Cd, roots absorbed 235.23 μg Cd/g root dry weight. Over the range of lnM to 5 μM Cd, exchangeable Cd represented approximately 5% of the absorbed fraction, and about 25 % of the total absorbed Cd was adsorbed to the root. Cadmium was passively absorbed to about 30% as observed in the presence of the inhibitor (DCCD). Ative absorption which represented 70% of Cd uptake involved H⁺‐ATPase carriers. Cadmium absorption was reduced to 30 and 20% in presence of lanthanum (La³⁺) and zinc (Zn²⁺), respectively which suggested that calcium (Ca), Cd, and Zn use the same carriers. Cadmium uptake in presence of DNP or NaN3 was approximately 4‐ fold that in control. Data showed presomption for an excretion of Cd out of root cells which could be the expression of a detoxification process limiting cell contamination.     

Host/parasite relationship in the in vitro infection of rat gliocytes by Neospora caninum: evaluation of cell respiration

Neospora caninum is a protozoon that causes abortion in cattle and neuromuscular lesions in dogs, with the formation of cysts mainly in the central nervous system. Since N. caninum is an intracellular parasite with tropism for the cells of nervous system, this study evaluated the respiratory metabolism of glial cells infected by this* parasite. Glial cultures obtained from the cerebral cortex of newborn rats were kept in DMEM enriched with 10% fetal bovine serum, 1 mM pyruvic acid and 2 mM of L-glutamine. They were infected at a ratio of approximately 1:1 (cell/parasite). Oxygen consumption was evaluated by polarography in the non infected and N. caninum infected groups, 24 and 72 h following infection. Glial cell respiration after 24 and 72 h was 307.2 +/- 34.7 and 308.9 +/- 64.1 microL of oxygen per mug of total protein per minute, and 566.2 +/- 54.6 and 579 +/- 117.5 microL O2/microg of total protein/minute in the control and infected groups, respectively. These results show that N. caninum does not interfere with glial respiration in vitro.     

Variation of Phyllosphere Microflora of Different Rice Varieties in Sri Lanka and its Relationship to Leaf Anatomical and Physiological Characters

Anatomical and physiological characters of the leaf surface and its physico‐chemical environment substantially influence the density and diversity of phyllosphere‐inhabiting microorganisms, which may include natural antagonists of important pathogens. The objective of this investigation was to quantify the phyllosphere (i.e. leaf surface) microbial population in a range of rice varieties grown in Sri Lanka and to identify the leaf anatomical and physiological characters that determine the density and diversity of phyllosphere microbes. Fifteen rice varieties including both traditional and new high‐yielding varieties were used in a planthouse experiment and a field experiment in two consecutive seasons to quantify the phyllosphere microbial population and measure leaf characters that may influence it. There were highly significant intervarietal variations in the density and diversity of epiphytic bacterial, fungal and total microbial populations under both planthouse and field conditions. However, there was no difference between traditional and new, high‐yielding varieties in their capacity to harbour epiphytic microbes in the phyllosphere. Total microbial density (TMD) under both conditions showed positive correlations with leaf hair density, stomatal density and transpiration rate. Under planthouse conditions, TMD was also positively correlated with leaf hair length and negatively correlated with leaf temperature. These correlations can be explained in terms of providing favourable microsites on the phylloplane for epiphytic microbial growth.