Effect of human disturbance on the structure and regeneration of forests in the Nevado de Toluca National Park, Mexico

Sample plots were established in the principal forest types in the the Nevado de Toluca National Park, Mexico including those dominated by Pinus hartwegii, Abies religiosa, Quercus laurina and Alnus jorullensis. The vertical structure was defined by three strata in the coniferous forests and two strata in the broadleaved forests. Timber harvesting in Abies religiosa and Quercus laurina forests and fires generated by humans in Pinus hartwegii forests impeded the recruitment of saplings. Mature trees were also heavily impacted by logging in Pinus hartwegii forests. On the contrary, Alnus jorullensis forests were increasing due to the disturbance of Pinus and Quercus forests, as well abandoned crop lands within the park. A combination of logging, uncontrolled fire, and grazing appears to be compromising the recruitment of important tree species in this national park. These factors, together with human settlements, have also increased the proportion of early successional species. Changes in forest structure from human disturbance indicate a need to control these activities if conservation goals are not to be compromised.     

Pyrosequencing of the rabies virus glycoprotein gene to demonstrate absence of vaccine-associated rabies cases following oral vaccination

Replication competent vaccines have been used successfully for the control of terrestrial rabies, mainly in wildlife; however, these vaccine strains occasionally may induce rabies. In this study, a pyrosequencing protocol for the rapid identification of vaccine-associated rabies viruses was applied to the 2008–2011 Italian epidemic. There was no evidence of vaccine-associated rabies cases following oral vaccination of foxes with the SAG2 and SADB19 vaccine strains.     

A rare Cryptosporidium parvum genotype associated with infection of lambs and zoonotic transmission in Italy

An outbreak of cryptosporidiosis occurred in a mixed sheep/cattle farm of Central Italy in October 2011. A total of 450 ovines (250 sheep and 200 lambs) and 140 bovines (130 cows and 10 calves) were housed in two separated units, at the time of the outbreak. About half of the lambs had diarrhea due to Cryptosporidium sp. with a mortality rate of 80%; calves were not infected. Genomic DNA was extracted from an archived slide and from fecal specimens, and the parasite was identified as Cryptosporidium parvum by PCR and sequence analysis at the CpA135 gene. Genotyping at the GP60 gene showed the presence of a very rare genotype, IIaA20G2R1. Shortly after the outbreak was identified, the son of the farm's owner, aged 18 months, experienced an acute gastroenteritis and was hospitalized due to recurrent episodes of diarrhea, fever, vomiting and lack of appetite. The feces tested negative for bacteria and viruses, whereas cryptosporidiosis was diagnosed by microscopy and an immunochromatographic test. Molecular typing identified the C. parvum genotype IIaA20G2R1 in the feces of the child. This is the first case of transmission of cryptosporidiosis in Italy involving lambs as source of oocysts infectious to humans.     

Effects of α-tocopherol and Ascorbic Acid on Equine Semen Quality after Cryopreservation

This study investigated the effects of ascorbic acid and α-tocopherol supplementation on semen quality parameters of equine thawed-frozen semen. Semen was divided in seven different treatments in a final concentration of 100 × 10⁶ sperm/mL by using Gent extender containing no supplements (control) and the following supplements with three different concentrations: α-tocopherol (0.5, 1, and 2 mM) and ascorbic acid (0.45, 0.9, and 1.8 g/L). After thawing, all samples were maintained at 37°C, while analyses were performed at 0, 60, and 120 minutes. Evaluation of viability and acrosome status (using Pisum sativum agglutinin conjugated to fluorescein isothiocyanate and propidium iodide), mitochondrial membrane potential (5,5′,6,6′-tetrachloro-1,1′,3,3′tetraethylbenzimidazolyl carbocyanine iodine [JC-1]), membrane lipid peroxidation (LPO; C₁₁-BODIPY^581/591), and stability of the plasmatic membrane (merocyanine 540 and Yo-Pro-1) of each sample was determined by flow cytometry. Relative to the control group, supplementation with α-tocopherol improved (P ≤ .05) postthaw membrane LPO, yet the higher concentrations of ascorbic acid (0.9 and 1.8 g/L, respectively) showed a negative effect on membrane LPO. Neither antioxidant significantly increased (P > .05) the acrosome integrity and mitochondrial membrane potential of frozen-thawed spermatozoa, although supplementation with α-tocopherol and ascorbic acid (0.9 and 1.8 g/L, respectively) had a positive effect on membrane integrity and stability (P ≤ .05). For all semen parameters, the lower concentration of ascorbic acid (0.45 g/L) did not show significant differences (P > .05) compared with the control. In conclusion, α-tocopherol seems to be an efficient antioxidant for reducing the oxidative stress provoked by cryopreservation, decreasing lipid peroxidation on equine spermatozoa.     

Meat quality and nutritional composition of pheasants (Phasianus colchicus) reared in an extensive system

Abstract 1. Carcass cuts, chemical composition, colour, textural parameters, fatty acid and amino acid profiles, macro- and micromineral content and sensory properties of breast and drumstick meat from 10 month old common pheasant produced in an extensive rearing system were analysed.

2. Breast muscle was characterised by a high concentration of protein (254 g/kg), low content of intramuscular fat (1.3 g/kg), considerable amount of Fe-haeme (4.9 mg/kg) low shear force (1.96 kg/cm²) and hardness (4.07 kg/cm²).

3. The main drumstick characteristics were 222 g of protein/kg, 4.0 g of intramuscular fat/g and a significant higher cholesterol concentration compared to that of breast (7.7 vs. 2.9 g/kg).

4. Saturated, monounsaturated and polyunsaturated fatty acid mean values for both muscle types were approximately 34.5%, 43.3% and 24.2% of total fatty acid methyl esters, respectively, and n-3 content in breast was significantly higher (3.21% vs. 1.65%). Significant differences were found between the three tissues (breast, drumstick and subcutaneous fat).

5. Large differences in the amino acid profile between breast and drumstick were detected, especially in the non-essential fraction. The main essential amino acids were lysine and leucine, whereas aspartic and glutamic acids were the most important non-essential amino acids.

6. There were significant differences between muscle types in the mineral content with regard to iron, magnesium, sodium and zinc. Pheasant meat is a good nutritional source of iron as 100 g of drumstick may provide 23.6% of the recommended daily amount.     

Grass silage for biorefinery—A meta‐analysis of silage factors affecting liquid–solid separation

This meta‐analysis based on 19 studies from Finland comprising 43 grass silages was undertaken to evaluate the effect of silage quality on liquid yield, liquid composition and retained compounds in liquid using four different liquid–solid separation methods. Silages were classified according to species (grass, clover or a mixture of them), additive treatment (no, biological or formic acid‐based additive) and harvest (primary growth or regrowth). A mixed model regression analysis with random study effect was used to evaluate the impact of silage characteristics on biorefinery efficiency. There was a large variation in silage quality in the data set. Silage dry‐matter concentration was the characteristic most highly correlated with liquid yield for all separation methods, and when used as an independent variable in the model, it resulted in the best predictions. The liquid–solid separation methods presented a great variation in the liquid yield, ranging from 0.26 to 0.56 when silage dry‐matter concentration was standardized to 250 g/kg. There was no effect of additive treatment and harvest in the estimation of the biorefinery potential, but species was a significant variable in predicting liquid yield for the laboratory‐scale presses with higher liquid yield for mixed grass and legume. The high correlation between silage quality and liquid yield and liquid composition provides potential to predict the biorefinery potential based on equations developed for each separation method. This information can be used to modify the silage production systems so that they best meet the requirements of a green biorefinery process.     

Ecotoxicological evaluation of Harbour sediments using marine organisms from different trophic levels

Background, Aim and Scope  The toxicity of contaminated sediments should be evaluated considering the direct exposure of laboratory organisms to whole sediments and the indirect exposure to elutriates or extracts (Tay et al. 1992, Byrne and Halloran 1999, Nendza 2002). The alga Dunaliella tertiolecta is indicated for the use in toxicity bioassays because it is highly sensitive to several xenobiotics. Harpacticoid copepods have been already used for toxicity testing and Tigriopus fulvus is a promising Mediterranean target-species in ecotoxicology (Todaro et al. 2001, Faraponova et al. 2003, Pane et al. 2005a). In this study, the toxicity of sediments collected in harbour sites of the Northeastern Adriatic Sea was evaluated by growth inhibition test with free living and alginate-immobilized Dunaliella tertiolecta and acute toxicity test with nauplii and adult Tigriopus fulvus with the aim of pointing out the importance to utilize model organisms from different trophic levels in sediment ecotoxicology. Methodology  Elutriates and whole sediments were tested on free living and immobilized (Pane et al. 1998) algal cells, and on laboratory reared copepods. Free-living D. tertiolecta were exposed to diluted elutriates in a static, multi-well plate system. Naalginate immobilized D. tertiolecta were placed in polystyrene inserts fitted with polyester mesh bottoms and exposed to a thin layer (2 mm) of whole sediments in multi-well plates (EPS 1992, Pane and Bertino 1999). Toxicity tests with copepods were carried out on Tigriopus fulvus nauplii (elutriates) and adults (whole sediments and elutriates). Same-aged nauplii useful for toxicity tests were obtained by egg sac detaching and consequent hatching stimulation (Pane et al. 2006). Newborn nauplii (I–II stage) were exposed to elutriates in multi-well plates provided with polystyrene inserts. Adult T. fulvus maintained in polystyrene inserts fitted with polyester mesh bottoms were placed in contact with a thin layer (2 mm) of whole sediment placed on multi-well plate bottoms. All end-points were evaluated after 96 h. Results  In general, the effects increased with the increasing of elutriate concentration up to 50%; the stimulation or inhibition of algal growth was statistically significant in comparison to the control. The inhibiting elutriates induced EC₅₀ variations of algal growth ranging from 66.9% to 74.3%. The mortality of T. fulvus nauplii was always < 25% after treatment with 100% elutriates and < 10% after treatment with 50% dilution; no effect was shown up with 25% dilution; therefore LC₅₀ was not calculable. The effect of elutriates was negligible on adult copepods and LC₅₀ values were never calculable; percent mortality always resulted in < 10% after treatment with whole sediments. Discussion  Both experimental systems gave substantially similar results after exposition to whole sediments and elutriates. During the experiment with algal cells, the immobilization in Na-alginate and the employment of inserts which allowed the contact of organisms with sediments and their easy counting were particularly useful. Likewise, the employment of inserts of adequate mesh size in the tests with copepods allowed the contact of organisms with the sediment and made organism handling and counting easy, as well as the evaluation of mortality. The methodology here described and the utilization of the proposed test-species could have an importance also considering that the current trend in ecotoxicological research is towards finding the most appropriate organism for specific areas of concern by using indigenous species (Mariani et al. 2006) and towards the major significance of chronic and reproductive end-points. Conclusions  Based on the above results, it can be stated that the bioassay with Dunaliella tertiolecta could be a good estimation tool for the ecotoxicological assessment of marine sediments. The immobilization of algae in Na-alginate was seen to be useful to evaluate the toxicity of whole sediments; the employment of polystyrene inserts allowed an improvement of the procedures. T. fulvus nauplii and adults, as other harpacticoids such as Tigriopus japonicus (Yoon et al. 2006), satisfy the basic criteria for the employment of a standard species in marine bioassays. To date only pelagic Acartia tonsa are utilized in the standardized procedure to evaluate the risk assessment of chemicals or wastewaters (ISO 1999). As, on the contrary, the exposure of copepods to solid-phase contaminants it is not yet standardized, the employment of polystyrene inserts improved the procedures for T. fulvus too. So, the rapidity and the possibility to solve practical problems could be the main attractive features of this technique (Pane et al. 2005a) when applied to whole sediments. Recommendations and Perspectives  The methodology here developed being also applicable to long term and reproduction tests should be recommended because it provides relevant information in comparison with other frequently applied, standardized biotests with crustaceans (ISO 1999). The procedure has been shown to be easily applicable to selected marine organisms. ESS-Submission Editor: Prof. Dr. Henner Hollert (henner.hollert@bio5.rwth-aachen.de)     

Identification of fructooligosaccharides in different banana cultivars

Banana has been currently indicated as a good source of fructooligosaccharides (FOS), which are considered to be functional components of foods. However, significant differences in their amounts in bananas have been observed in the literature. This work aims to identify and quantify FOS during ripening in different banana cultivars belonging to the most common genomic groups cultivated in Brazil. Considering that these differences can be due to cultivar, stage of ripening, and the methodologies used for FOS analyses, sugar contents were analyzed by high performance anion exchange chromatography-pulsed amperometric detection (HPAEC-PAD) and gas chromatography-mass spectrometry (GC-MS). An initial screening of eight cultivars (Ouro, Nanic?o, Prata, Ma??, Mysore, Pacovan, Terra, and Figo) in a full-ripe stage showed that 1-kestose, the first member of the FOS series (amounts between 297 and 1600 microg/g of DM), was accumulated in all of them. Nystose, the second member, was detected only in Prata cultivar. Five of the cultivars were analyzed during ripening, and a strong correlation could be established with a specific sucrose level ( approximately 200 mg/g of DM), which seems to trigger the synthesis of 1-kestose (the low amounts of FOS, below the functional recommended dose, indicates that banana cannot be considered a good source of FOS).     

Classification of Peruvian highland maize races using plant traits

The maize of Latin America, with its enormous diversity, has played an important role in the development of modern maize cultivars of the American continent. Peruvian highland maize shows a high degree of variation stemming from its history of cultivation by Andean farmers. Multivariate statistical methods for classifying accessions have become powerful tools for classifying genetic resources conservation and the formation of core subsets. This study has two objectives: (1) to use a numerical classification strategy for classifying eight Peruvian highland races of maize based on six vegetative traits evaluated in two years and (2) to compare this classification with the existing racial classification. The numerical classification maintained the main structure of the eight races, but reclassified parts of the races into new groups (Gi). The new groups are more separated and well defined with a decreasing accession within group × environment interaction. Most of the accessions from G1 are from Cusco Gigante, all of the accessions from G3 (except one) are from Confite Morocho, and all of the accessions from G7 are from Chullpi. Group G2 has four accessions from Huayleño and four accessions from Paro, whereas G4 has four accessions from Huayleño and five accessions from San Geronimo. Group G5 has accessions from four races, and G6 and G8 formed small groups with two and one accession each, respectively. These groups can be used for forming core subsets for the purpose of germplasm enhancement and assembling gene pools for further breeding.