Impact of vertebrate acorn- and seedling-predators on a Mediterranean Quercus pyrenaica forest

We have experimentally investigated the impact of biotic factors, acting at the seed and seedling stages, on a Quercus pyrenaica forest in the Sierra Nevada mountains (SE Spain). We monitored the natural establishment of the oak for 3 years in two forest plots and two shrubland plots, by counting seedlings and juveniles. In addition, we established several experiments in these plots to examine acorn and seedling survival, while also considering the microhabitat effect on survival probability. Dispersed acorns were quickly consumed by several species of predators, particularly wild boar (Sus scrofa) and woodmouse (Apodemus sylvaticus). Less than 4% of the experimental acorns survived to produce seedlings, even when they were buried 4 cm in soil, simulating caches. No effect of microhabitat was found on predation, and thus no safe site appears to exist for Q. pyrenaica acorns in the study area. Some 98% of the 1000 experimental seedlings were killed by herbivores, notably woodmice, wild boar, and domestic and wild ungulates. Seedling survival varied spatially, being significantly higher under shrubs (4%) than in any other microhabitat (less than 0.5%). Both acorn and seedling survival were much lower in the shrublands than in the forests. In shrubland plots, the main agent of seedling mortality was trampling by domestic ungulates foraging in herds. This study suggests that the regeneration of Q. pyrenaica forests in Mediterranean mountains can be limited by herbivores acting at several life-history stages.     

Modeling cotton production response to shading in a pecan alleycropping system using CROPGRO

Light optimization assessment in alleycropping systems through model application is becoming an integral part of agroforestry research. The objective of this study was to use CROPGRO-cotton, a process-based model, to simulate cotton (Gossypium hirsutum L.) production under different levels of light in a pecan (Carya illinoensis K. Koch) alleycropping system in Jay, Florida, USA. Soil classification in the area was Red Bay sandy loam soil (Rhodic Paleudult). To separate roots of cotton and pecan, polyethylene-lined trenches were installed parallel to tree rows, thus competition for water and nutrients was assumed to be non-existent. Four treatments were set up in the CROPGRO-cotton model, as follows: (1) control (full amount of light transmittance), (2) Row 1 (50% light transmittance), (3) Row 4 (55% light transmittance), and (4) Row 8 (70% light transmittance). Cotton model parameters affecting specific leaf area (SLA), leaf area index (LAI), maximum leaf photosynthetic rate (FLMAX) and carbon partitioning were calibrated using the full sun treatment. Measurements of SLA, LAI, and aboveground biomass were made on the different shaded treatments and compared with simulated values. Simulation results showed that aboveground mechanisms affecting production in shaded environment (i.e., SLA, LAI, LFMAX, and carbon partitioning) influence model behavior. After calibration, the model predicted SLA of cotton in all treatments with reasonable precision. However, LAI was underestimated in the more shaded treatment rows 4 and 8. Generally, the model provided a close agreement between measured and simulated biomass both in 2001 and 2002 (R ² = 0.95 and R ² = 0.92, respectively). In 2001, predicted biomass for the control was 5,401 kg ha⁻¹ compared to the measured value of 5,393 kg ha⁻¹. A similar trend was also observed in 2002. The CROPGRO-Cotton model was able to describe variations in growth among the shaded treatments well across both growing seasons. However, it was found that additional research is needed to improve the model’s ability to simulate LAI under shading conditions. Parameters associated with photosynthesis and dry matter partitioning were reasonably stable across shading treatments and years but those associated with leaf area growth varied.     

Effects of temperature on early development of the New Zealand geoduck Panopea zelandica (Quoy & Gaimard, 1835)

Ambient seawater temperature is an important factor during the early life stages of marine invertebrates. Temperature is often manipulated in hatcheries to shorten the incubation period before the larval rearing phase. In this study, the effect of temperature on the early development of the geoduck Panopea zelandica was investigated over a 48‐hr period to identify the optimum temperature for fertilization and development in a controlled environment. Eggs and sperm collected from broodstock were exposed to ten temperatures ranging between 11.8 and 23.7°C, and fertilization and subsequent development were monitored over 48 hr. Highest percentages of fertilization were achieved at 23.7°C, which was the highest temperature tested in this study. However, the development of P. zelandica embryos was greatly hindered at temperatures >18.5°C due to a range of abnormalities arising from uneven cell division and cellular blebbing. All larvae died at the highest temperature of 23.7°C within 48 hr of exposure. The combined fertilization success and embryo development data indicate that 18.5°C is the optimal temperature for incubating P. zelandica embryos under hatchery conditions.     

Determination of epsilon-N-pyrrolylnorleucine in fresh food products.

epsilon-N-Pyrrolylnorleucine was determined in different fresh food products to study its presence as a normal component of food proteins. Twenty-two different products were screened: cod, cuttlefish, salmon, sardine, trout, beef, chicken, pork, broad bean, broccoli, chickpea, garlic, green pea, lentil, mushroom, soybean, spinach, sunflower, almond, hazelnut, peanut, and walnut. Foods were homogenized, their proteins were precipitated with trichloroacetic acid and hydrolyzed with 2 N NaOH for 20 h, and the epsilon-N-pyrrolylnorleucine content was determined by capillary electrophoresis. The epsilon-N-pyrrolylnorleucine, which was identified by HPLC/MS in sardine muscle hydrolysate, ranged in the 22 foods analyzed from 0.24 to 6.36 micromol/g. This concentration was correlated with the protein content of the food (r = 0.687, p = 0.00041). In addition, the epsilon-N-pyrrolylnorleucine/lysine ratio was found to be a function of the lipid, iron, and protein contents of the food (r = 0.881, p < 0.0001) and was directly correlated with lipid and iron contents and inversely correlated with the protein content. These results are in agreement with the oxidative stress origin proposed for epsilon-N-pyrrolylnorleucine and suggest that the epsilon-N-pyrrolylnorleucine/lysine ratio is a characteristic of each food. In addition, epsilon-N-pyrrolylnorleucine seemed to be a normal component of many fresh food products, in which it may be acting as a natural antioxidant.     

A highly sensitive and specific gel-based multiplex RT-PCR assay for the simultaneous and differential diagnosis of African swine fever and Classical swine fever in clinical samples

The development and standardisation of a novel, highly sensitive and specific one-step hot start multiplex RT-PCR assay is presented for the simultaneous and differential diagnosis of African swine fever (ASF) and Classical swine fever (CSF). The method uses two primer sets, each one specific for the corresponding virus, amplifying DNA fragments different in length, allowing a gel-based differential detection of the PCR products. Universal detection of ASF and CSF virus strains was achieved through selection of primers in conserved viral genome regions. The detection range was confirmed by analysis of a large collection of isolates of the two viruses. The high specificity of the assay was proven by testing related viruses, uninfected cell line cultures and healthy pig tissues. Additional confirmatory tests of the ASF and CSF virus amplicon specificity, based on restriction endonuclease analysis with BsmA I or Ban II, respectively, are also described. The analysis of whole blood and serum samples from experimentally infected animals proved the usefulness of the method for an early diagnosis of both diseases, even before the appearance of the first clinical signs. A study of 150 positive field samples from several ASF and CSF outbreaks showed the suitability of this method for a rapid (less than five hours), sensitive and specific differential diagnosis in clinical samples. In addition, a highly sensitive and specific uniplex RT-PCR for CSFV was also developed and standardised as a powerful tool for fast and early diagnosis of the disease.     

Water diversion reduces abundance and survival of two Mediterranean cyprinids

Water flow is a fundamental characteristic required for the ecological integrity of stream ecosystems. However, populations of many freshwater fishes in the Mediterranean region are threatened by man‐induced drought due to water diversion. Mark‐recapture methodology is an effective tool for estimating fish abundance and survival probability, but it has been seldom used with Mediterranean freshwater fish. We tagged over 2,400 individuals of two threatened cyprinids (Mediterranean barbel Barbus meridionalis and Catalan chub Squalius laietanus) inhabiting an Iberian stream affected by water diversion, and used mark‐recapture methods to evaluate the effects of flow, temperature and depth on their apparent survival and abundance at hydrologically altered and perennial reaches. Based on estimates over the summer drying season, survival of Mediterranean barbel was more than six times higher in an upstream reach with permanent flow than a middle reach impacted by water diversion. Water depth was the most important habitat feature accounting for differences in survival, with both barbel and chub exhibiting higher survival and abundance at permanent sites farther downstream where flow had been restored and in an unaltered tributary. Our results show clear negative impacts of water diversion on fish populations. Both species, but particularly barbel, have life‐cycle characteristics that may allow populations to recover quickly if natural streamflow was restored by conserving water and reducing water diversion.     

Conjunctival vaccination of the brushtail possum (Trichosurus vulpecula) with bacille Calmette-Guérin

AIM: To determine the efficacy of conjunctival vaccination of captive brushtail possums (Trichosurus vulpecula) with bacille Calmette-Guérin (BCG), as measured by immunological responses to vaccination and response to intratracheal challenge with Mycobacterium bovis.

METHODS: Nine adult male brushtail possums were vaccinated by the instillation of a suspension of BCG strain Pasteur 1173P2 into the conjunctival sac of each eye. Each drop contained approximately 2.5 × 10⁵ colony forming units (cfu). At 8 weeks post-vaccination (pv) the vaccinated possums and 10 unvaccinated possums were challenged by intratracheal instillation of approximately 100 cfu of M. bovis. Cellular immune responses to bovine purified protein derivative (PPD) antigen were measured using the lymphocyte proliferation assay (LPA). Possums surviving to 50–51 days after challenge were euth anised and subjected to detailed post-mortem examination, including histopathology, to assess protection against tuberculosis. Sections of lung and spleen were cultured for M. bovis.

RESULTS: No conjunctival inflammation or other adverse reactions to the administration of the vaccine were evident macroscopically. The vaccinated group showed a systemic cellular immune response to bovine PPD antigen at 4 and 8 weeks pv, and the response at 8 weeks was significantly greater than at 4 weeks (p<0.05). Conjunctival vaccination induced significant levels of protective immunity, measured as less mass of tuberculous lesions in lung (p<0.05) and less dissemination of disease in vaccinated compared with unvaccinated possums (p<0.05).

CONCLUSIONS: Conjunctival vaccination with BCG induced a significant level of protective immunity against M. bovis infection in possums. This route of vaccination, together with intranasal aerosol vaccination, could be utilised in the delivery of an aerosolised vaccine using a device that sprays the vaccine suspension into the eyes and nose of possums.     

Growth of Hatchery‐reared Juveniles of the Australasian Sea Cucumber,Australostichopus mollis,Fed with Artificial and Natural Diets

Australostichopus mollis is the most common and valuable sea cucumber species on the coasts of New Zealand and southern Australia. However, information supporting the development of artificial diets to advance the aquaculture of this species is scarce. In this study, the nutrient absorption, growth, pigmentation, and survival of small (111 mg) and large (434 mg) early juveniles were measured. The juveniles were fed with mussel biodeposits (MB); a commercial hatchery diet‐powdered macroalgae (HD); and artificial diets with different protein levels, 5% (5P) and 10% (10P), respectively. During the experiment, survival was greater than 66% in all diet treatments, with the juveniles feeding actively and absorbing all major macronutrients. The small juveniles, which were starting pigmentation, had higher growth rates (0.8–2.5%/d) than the large juveniles (0.2–1.2%/d), which were completing pigmentation. Growth was higher with the MB and HD diets than with the 5P and 10P diets, which could be explained by differences in the food consumption and nutrient absorption among the diet treatments. This study shows the feasibility of developing artificial diets without depending on naturally available ingredients; however, palatability and consumption need to be improved in order to make artificial diets for this species more effective and commercially available.