Variation in clearance and ingestion rates by larvae of the black-lip pearl oyster (Pinctada margaritifera, L.) feeding on various microalgae

Clearance rate (CR) and ingestion rate (IR) of different sizes (89, 125 and 188 μm shell length) of Pinctada margaritifera larvae were determined when feeding on various microalgae. The microalgae tested were the diatoms, Chaetoceros muelleri and C. simplex, and flagellates, Tahitian Isochrysis aff. galbana, Pavlova lutheri and P. salina at 5 or 10 cells μL^–1. Both CR and IR of microalgae tested in this study increased with increasing larval size; but at all larval sizes, diatoms resulted in lower CR and IR. Of the microalgae tested, P. margaritifera larvae showed greatest CR and IR with the two Pavlova spp. Maximum CR for P. salina was 10.5, 21.2 and 29.7 μL h^–1 for larvae with shell lengths of 89, 125 and 188 μm, respectively. The highest IR values for P. margaritifera larvae with shell lengths of 89, 125 and 188 μm were 8.7, 81.0 and 165.7 cells·larva^–1 h^–1, respectively. CR and IR of P. salina were approximately five times higher than those recorded for C. muelleri and C. simplex.     

Reserve effect of a small North-East Atlantic marine protected area (Arrábida,Portugal) on soft-sediment fish species

1. The implementation of marine protected areas (MPAs) has been widely used as a tool to manage and conserve marine resources and services. Yet, to date, the reserve effect is still weakly evaluated, particularly for soft-sediment habitats.
2. The Arrábida MPA, considered as a biodiversity hotspot, was fully established in 2009 on the west coast of Portugal and is characterized by large expanses of soft-sediment habitats. This MPA was established to protect biodiversity and to ensure the livelihood of the local small-scale fisheries community.
3. Beyond before–after-control–impact (BACI) analyses were carried out on catch data (abundance and biomass) of 351 trammel net sets from experimental fishing campaigns (2007–2019) to study the reserve effect on demersal fish populations.
4. The results show a declining trend in abundance inside and outside the protected area, with significant positive effects only found for undersize commercial species and Solea senegalensis, and a general increase in fish species total length. Despite the lack of any obvious reserve effect, the increase in fish length can be considered as a first sign.
5. Anthropogenic pressures are generalized around the area, and transgressions of the restrictions are frequently observed inside the no-take zone. These can, among other aspects, delay or prevent the expected reserve effects. Therefore, enforcement and/or modification of the spatial plan must be incentivized to achieve the goals of the MPA.

     

The influence of the tracer injection–acquisition interval on the distribution of 99mTc-ECD in the brain of laboratory cats

Single-photon emission computed tomography (SPECT) can be used to study the regional distribution of the cerebral blood flow in the feline brain without interference of anesthetics. When the tracer, ^99mtechnetium-ethyl cysteinate dimer (^99mTc-ECD), is injected in the awake animal, the acquisition can be performed afterward under general anesthesia, whereas the tracer distribution still represents the regional cerebral blood flow (rCBF) in the awake animal. The aim of this study was to look at the in vivo stability of ^99mTc-ECD in the feline brain. For this purpose, 6 cats (n = 6) were used, and 3 serial acquisitions were performed starting at 40 (T40), 60 (T60), and 85 (T85) minutes after tracer injection. Total counts and perfusion indices (PIs), normalized to the total brain and to the cerebellum, were calculated. When T85 was compared with T40, total counts decreased, depending on the brain region, with 29% (left thalamus) to 51% (bulbus olfactorius). These regional differences in tracer clearance resulted in significantly altered PIs at T85 as compared with T40. This study shows that ^99mTc-ECD SPECT can be used in cats for studying the rCBF and that the clearance of ^99mTc-ECD in the feline brain is region dependent. As a result, the acquisition should be started between 40 and 60 minutes after tracer injection.     

Species-specific DNA markers for identification of two root-knot nematodes of coffee: Meloidogyne arabicida and M. izalcoensis

Seven root-knot nematodes (RKN), including Meloidogyne exigua, M. incognita, M. paranaensis, M. enterolobii, M. arabicida, M. izalcoensis and M. arenaria are major pathogens of coffee crop in the Americas. Species-specific primers for their identification have been developed for five of them and constitute a fast and reliable method of identification. Here we report a PCR-based assay for specific detection of M. arabicida and M. izalcoensis. Random Amplified Polymorphic DNA fragments specific for these two species were converted into sequence characterized amplified region (SCAR) markers. PCR amplification using the SCAR primers produced a specific fragment of 300 bp and 670 bp for M. arabicida and M. izalcoensis, respectively, which were absent in other coffee-associated Meloidogyne spp. tested. SCAR primers also allowed successful amplification of DNA from single second-stage juveniles (J2), males and females. In addition, these primers were able to unambiguously detect the target species in nematode suspensions extracted from soil and roots samples, in different isolates of the same species or when used in multiplex PCR reactions containing mixtures of species. These results demonstrated the effectiveness of these SCAR markers and their multiplex use with those previously developed for M. exigua, M. incognita, M. paranaensis, M. enterolobii and M. arenaria constitute an essential detection tool. This diagnostic kit will contribute for specific J2 identification of the major RKN infecting coffee from field samples in the Americas.     

Assessment of a pyrogallol red technique for total protein measurement in the cerebrospinal fluid of dogs

The measurement of protein concentration in the cerebrospinal fluid is a basic analytical method in neurology. In this study, a pyrogallol red technique using a human albumin calibrator previously validated in human medicine was tested for canine samples, and the results were compared with those obtained using urine test strips. Pyrogallol red significantly (P<0.05) but moderately underestimated purified dog albumin and globulins. The imprecision of the technique was low: intra- and between-series coefficients of variation were 1.6 and 4.3 per cent at protein concentrations of about 0.3 g/litre. Over 49 samples, there was good agreement between the pyrogallol red and test strip results (r=0.63), especially for low and high protein concentrations, but misclassifications were observed with '+' test strip readings.     

Response of black rot in cabbage to spatial distribution of inoculum

Disease progress of black rot in cabbage crop was studied over three years in field plots to compare the effects of uni-focal and multi-focal inoculum applied in equal amounts per plot. Disease progress (plant incidence and leaf incidence) was plotted over time, three dimensional maps were made, and disease aggregation was studied by means of geostatistics, black-black counts and Moran's I statistic. Black rot progress was primarily due to focus expansion. Secondary foci may appear at short distances from the initial focus but they usually merge with the expanding initial focus. Anisotropy occurred occasionally but was of minor importance. Disease proceeds faster in plots with multi-focal inoculation than in those with uni-focal inoculation. Probably, serious epidemics in Dutch cabbage fields originate from large numbers of foci.     

Genetic variability of rhizobacteria from wild populations of four Lupinus species based on PCR-RAPDs§

The metabolic capacities of rhizosphere bacteria can depend on intraspecific genetic variability at strain level. We sampled bacteria from the rhizosphere of three populations of four different Lupinus species at two growth stages (flowering: GS1 and fruiting: GS2). Isolates were identified to the genus level by classical biochemical tests. The most abundant genera found were Bacillus, Aureobacterium, Cellulomonas, Pseudomonas, and Arthrobacter. Genetic divergence of rhizobacteria was tested by PCR-RAPDs. The genetic distances were low, with mean values of 37 % for Bacillus, 25 % for Aureobacterium, 46 % for Cellulomonas, 16 % for Pseudomonas, and 23 % for Arthrobacter. Aureobacterium, the most abundant genus, predominated in the rhizosphere of all populations and at both growth stages (GS1 and GS2) of L. angustifolius. The Aureobacterium strains consisted of 11 groups with 90 % similarity indexes. The cluster analysis of these groups shows that strains isolated from different lupin species and sampling times have extraordinary low diversity indexes, or are even identical. This fact, together with the low genetic distance detected in the rhizosphere, reveals a clear specificity in the plant-bacteria interaction. This specificity could be related with several aspects of plant physiology.     

Broodstock spawning and larviculture of whiting (Merlangius merlangus L.) reared in captivity

In recent years, an increasing interest in the farming of marine ‘white’ fish, such as cod, has been observed in the aquaculture industry. One species being considered for aquaculture development is whiting (Merlangius merlangus L.). With little data being available on the domestication of this species, our study aimed to characterize spawning behaviour and rhythmicity in captive broodstock and provide original data on egg incubation, embryonic development and larval rearing. The results observed through video recording showed that spawning mainly occurred at night with active courtship behaviour. Daily egg batches were collected from two separate spawning seasons, with an average fertilization success declining from 77% to 53%. Time to hatch was inversely related to the incubation temperature and occurred after 78 degree‐days (9 days at 8 °C), with emerging larvae ranging in size from 3.3 to 3.46 mm total length and hatch success ranging from 48.3% to 99.9%. In comparison with a clear water environment, a green water regime during the first days of larval rearing significantly improved larval growth and survival. These results are the first to describe spawning activity, early development and larval performances in whiting and are promising for the potential future development of whiting aquaculture.     

Multivariate analysis of toxicological and environmental properties of soil nematicides

BACKGROUND: In intensive agriculture, the use of pesticides and soil fumigants is necessary to produce economically viable crops worldwide. However, this practice may involve undesirable effects on human health and the environment. In 1995, methyl bromide was restricted by the Montreal Protocol because of possible ozone depletion. The objective of this study was to compare intrinsic environmental and toxicological properties of 11 active substances with nematicidal properties, some of them recognized as methyl bromide alternatives. RESULTS: Four groups of active substances were discriminated by a series of principal component analyses (PCAs): (a) high toxicity to non-target fauna, humans and animals and medium persistence in the environment (cadusafos, ethoprophos and fenamiphos); (b) high toxicity to humans, animals and non-target fauna and high persistence (carbofuran and fosthiazate); (c) low toxicity to non-target fauna, humans and animals and low persistence (carbosulfan, benfuracarb and oxamyl); (d) low toxicity to humans, animals and non-target fauna and medium persistence in the environment (1,3-dichloropropene, chloropicrin and methyl bromide). CONCLUSION: Evaluating the multiple aspects of toxicological and environmental properties of active substances through PCA is proposed as a helpful tool for initially comparing the complex toxicological behaviour of active substances as plant protection products.