Recirculation nursery systems for bivalves

In order to increase production of bivalves in hatcheries and nurseries, the development of new technology and its integration into commercial bivalve hatcheries is important. Recirculation aquaculture systems (RASs) have several advantages: high densities of the species can be cultured resulting in a cost-effective production system; optimal temperature maximizes production and allows rapid turnover of the product; stable water quality improves growth rate and minimizes stress and potential loss by diseases. Pilot RAS systems were developed for seed rearing of oysters (Crassostrea gigas), scallops (Pecten maximus), mussels (Mytilus edulis) and clams (Ruditapes decussatus). Optimal feed addition and waste matrix were determined. Based on this, system flow rates were designed. Seed growth in the pilot RAS systems was compared at different renewal rates and with growth in flow-through systems (FTS). All four species can be reared in RAS and showed similar growth in RAS and in FTS or in RAS with a higher renewal rate. RAS can keep O₂, nitrogen and pH within the desired range. Temperature was generally higher in RAS than in FTS, probably due to heat induced by the pump circulating the water. The supply of sufficient amount of food in combination with a desire to reduce the renewal rate calls for use of concentrated feed in RAS.     

Understanding physical clogging in drip irrigation: in situ,in-lab and numerical approaches

Dripper clogging is a major drawback of microirrigation systems that must be addressed to improve their efficiency and durability. Particle-induced clogging is first studied in situ. The experiments consist in observing in real conditions the behavior of a series of drippers fitted on an agricultural plot in the south of France. The plot is supplied from a canal with Durance River water. The latter is loaded with sediments that gradually clog drippers and filters. Water analysis reveal that physicochemical clogging prevails over biological clogging. This characterization helps in setting in-lab experiment protocol. Indeed, besides field observation of clogging, laboratory analyses of both the irrigation water and the clogging material are performed with reactive and inert clay: smectite and an illite–calcite mix. A surprising tendency is observed: Salt concentration in smectite seeded water decreases the clogging, whereas it increases agglomerate size. Computational fluid dynamic simulations are carried out to investigate the impact of particles on flow behavior. Results demonstrate that clay particles interacting with the flow govern the complex structure of the fluid velocity fields inside the dripper labyrinth channel.     

Chemical characterization of wood and extractives of fast-growing Schizolobium parahyba and Pinus taeda

This study aims to evaluate the chemical composition of wood and extractives of Pinus taeda and Schizolobium parahyba (guapuruvu) as potential feedstock for new applications in the biorefinery industry. For this purpose, their content of α-cellulose, hemicellulose, insoluble lignin, hot water solubility, NaOH_1% solubility, inorganic materials (ash), and monomeric sugars by high-performance liquid chromatography was quantified. Attenuated total reflectance infrared spectroscopy and thermogravimetric analysis were also used to complete the physicochemical characterization of the studied woods. The extractives were obtained by soxhlet extraction with ethanol:toluene and dichloromethane and identified with pyrolysis-gas chromatography/mass spectroscopy technique. The results showed that guapuruvu wood has the higher amount of hemicellulose (16%) when compared to pine wood (10%), which resulted in higher solubility in alkali solution. Furthermore, in relation to other biomasses, the two woods presented more percentage of lignin and minor content of hemicelluloses. The P. taeda wood presented the highest percentage of extractives mainly composed of fatty acids and aromatic hydrocarbons, while guapuruvu wood had a higher percentage of phenolic compounds and also fatty acids. Both the materials have low content of extractives with dichloromethane and were mainly composed of lipophilic compounds.     

DNA Quantity and Quality in Remnants of Traffic-Killed Specimens of an Endangered Longhorn Beetle: A Comparison of Different Methods

The sampling of living insects should be avoided in highly endangered species when the sampling would further increase the risk of population extinction. Nonlethal sampling (wing clips or leg removals) can be an alternative to obtain DNA of individuals for population genetic studies. However, nonlethal sampling may not be possible for all insect species. We examined whether remnants of traffic-killed specimens of the endangered and protected flighless longhorn beetle Iberodorcadion fuliginator (L., 1758) can be used as a resource for population genetic analyses. Using insect fragments of traffic-killed specimens collected over 15 yr, we determined the most efficient DNA extraction method in relation to the state of the specimens (crushed, fragment, or intact), preservation (dried, airtight, or in ethanol), storage duration, and weight of the sample by assessing the quantity and quality of genomic DNA. A modified cetyltrimethyl ammonium bromide method provided the highest recovery rate of genomic DNA and the largest yield and highest quality of DNA. We further used traffic-killed specimens to evaluate two DNA amplification techniques (quantitative polymerase chain reaction [qPCR] and microsatellites). Both qPCR and microsatellites revealed successful DNA amplification in all degraded specimens or beetle fragments examined. However, relative qPCR concentration and peak height of microsatellites were affected by the state of specimen and storage duration but not by specimen weight. Our investigation demonstrates that degraded remnants of traffic-killed beetle specimens can serve as a source of high-quality genomic DNA, which allows to address conservation genetic issues.     

Low genetic variability in Sclerotinia sclerotiorum populations from common bean fields in Minas Gerais State,Brazil, at regional,local and micro‐scales

Sclerotinia sclerotiorum, causal agent of white mould, is the most destructive and widely distributed soilborne pathogen of common bean during the autumn–winter season in Brazil. Nevertheless, little is known about the genetic structure of the pathogen population. Microsatellite (SSR) markers and mycelial compatibility groups (MCGs) were used to characterize 118 isolates collected from 20 bean fields located in the most important growing regions of Minas Gerais State (MG). Additionally, the genetic variability among 10 isolates obtained from a single sclerotium was investigated in 10 different sclerotia. Seventy SSR haplotypes and 14 MCGs were identified among the 118 isolates. The genetic differences within bean growing areas accounted for most of the genetic variation (72%). Despite the relatively high genotypic diversity, the SSR loci were at linkage disequilibrium. Moreover, 70% of the isolates were assigned to only two MCGs, and haplotypes of a given MCG were closely related. The discriminant analysis of principal components revealed five groups. There was strong genetic differentiation between isolates collected in one municipality in southern MG when compared to other regions. Common bean resistance to white mould should be assessed with representative isolates of the five genetic groups and, if possible, of the different MCGs detected in the present study. One to five haplotypes were detected among the 10 isolates obtained from a single sclerotium. Therefore, in order to ensure genetic identity of an isolate, hyphal tip or monoascosporic isolates should be used.     

Genetic diversity of the root‐knot nematode Meloidogyne ethiopica and development of a species‐specific SCAR marker for its diagnosis

Meloidogyne ethiopica is an important nematode pathogen causing serious economic damage to grapevine in Chile. In Brazil, M. ethiopica has been detected with low frequency in kiwifruit and other crops. The objectives of this study were to evaluate the intraspecific genetic variability of M. ethiopica isolates from Brazil and Chile using AFLP and RAPD markers and to develop a species‐specific SCAR‐PCR assay for its diagnosis. Fourteen isolates were obtained from different geographic regions or host plants. Three isolates of an undescribed Meloidogyne species and one isolate of M. ethiopica from Kenya were included in the analysis. The results showed a low level of diversity among the M. ethiopica isolates, regardless of their geographical distribution or host plant origin. The three isolates of Meloidogyne sp. showed a high homogeneity and clustered separately from M. ethiopica (100% bootstrap). RAPD screenings of M. ethiopica allowed the identification of a differential DNA fragment that was converted into a SCAR marker. Using genomic DNA from pooled nematodes as a template, PCR amplification with primers designed from this species‐specific SCAR produced a fragment of 350 bp in all 14 isolates of M. ethiopica tested, in contrast with other species tested. This primer pair also allowed successful amplification of DNA from single nematodes, either juveniles or females and when used in multiplex PCR reactions containing mixtures of other root‐knot nematode species, thus showing the sensitivity of the assay. Therefore, the method developed here has potential for application in routine diagnostic procedures.     

Validation of two models for self‐incompatibility in autotetraploid perennial ryegrass using high resolution melting‐based markers

Perennial ryegrass (Lolium perenne L.) displays a two‐locus gametophytic self‐incompatibility (SI) system that remains intact at the tetraploid level. Two models are plausible for SI in autotetraploids. In Model I: both alleles at the S locus and both at the Z locus in diploid pollen matching the female genotype results in incompatibility. In Model II: only one allele at S and one at Z locus in diploid pollen matching the female results in incompatibility. The goals were to determine which of the models best explains SI in our autotetraploid ryegrass population and to evaluate the efficiency of high‐resolution melting (HRM) genotyping for discriminating different iso‐allelic genotypes. The progeny of a cross between two autotetraploids was characterized with three HRM‐based markers co‐segregating with Z. Segregation ratios were used to make inferences about the mode of action of the SI system. The observed segregation differed significantly (P < 0.001) from the expected under Model I, but not from the expected under Model II (P = 0.463). Thus, Model II explains SI in this population, and HRM is an efficient tool to distinguish different iso‐allelic genotypic classes.     

Ecological fitness and strategies of adaptation of Bartonella species to their hosts and vectors

Bartonella spp. are facultative intracellular bacteria that cause characteristic host-restricted hemotropic infections in mammals and are typically transmitted by blood-sucking arthropods. In the mammalian reservoir, these bacteria initially infect a yet unrecognized primary niche, which seeds organisms into the blood stream leading to the establishment of a long-lasting intra-erythrocytic bacteremia as the hall-mark of infection. Bacterial type IV secretion systems, which are supra-molecular transporters ancestrally related to bacterial conjugation systems, represent crucial pathogenicity factors that have contributed to a radial expansion of the Bartonella lineage in nature by facilitating adaptation to unique mammalian hosts. On the molecular level, the type IV secretion system VirB/VirD4 is known to translocate a cocktail of different effector proteins into host cells, which subvert multiple cellular functions to the benefit of the infecting pathogen. Furthermore, bacterial adhesins mediate a critical, early step in the pathogenesis of the bartonellae by binding to extracellular matrix components of host cells, which leads to firm bacterial adhesion to the cell surface as a prerequisite for the efficient translocation of type IV secretion effector proteins. The best-studied adhesins in bartonellae are the orthologous trimeric autotransporter adhesins, BadA in Bartonella henselae and the Vomp family in Bartonella quintana. Genetic diversity and strain variability also appear to enhance the ability of bartonellae to invade not only specific reservoir hosts, but also accidental hosts, as shown for B. henselae. Bartonellae have been identified in many different blood-sucking arthropods, in which they are typically found to cause extracellular infections of the mid-gut epithelium. Adaptation to specific vectors and reservoirs seems to be a common strategy of bartonellae for transmission and host diversity. However, knowledge regarding arthropod specificity/restriction, the mode of transmission, and the bacterial factors involved in arthropod infection and transmission is still limited.