Culture of Differentiated Adult Rabbit Auricular Chondrocytes

Chondrocytes dedifferentiate to a fibroblast‐like phenotype on plastic surfaces. Dedifferentiation is reversible if these cells are then cultured embedded in gels as alginate, agarose or collagen. Chondrocytes cultured in suspension on a non‐adherent surface are also known to form aggregates of differentiated cells. The knowledge of chondrocyte behavior in culture is relevant for tissue engineering purposes. In this report we describe a simple method to culture differentiated or redifferentiated rabbit auricular chondrocytes on plastic surfaces with a stable phenotype. When chondrocyte aggregates formed in suspension are next seeded on plastic surfaces, most of them attach to the plastic as round or polygonal cells, and this morphological differentiation, confirmed by the presence of type II collagen, is stable for long culture periods. We also report that the addition of aggregates to monolayer cultures of dedifferentiated chondrocytes results in their redifferentiation, as is shown by their morphological changes and the synthesis of type II collagen. Therefore, this simple method can be useful for the study of chondrocyte behavior on plastic surfaces and for redifferentiating previously proliferated chondrocytes in tissue engineering techniques. Furthermore, these results demonstrate that, in addition to culture conditions such as cell isolation method or cell‐density, chondrocyte behavior on plastic depends on the presence or absence of aggregates resulting from the dissociation process.     

Socio-spatial organization reveals paternity and low kinship in the Honduran white bat (Ectophylla alba) in Costa Rica

Ectophylla alba is a tent-making bat that roosts in mixed-sex clusters comprising adults and offspring. Our goal was to determine the genetic identity of individuals belonging to different roosting groups. We tested the hypothesis of kin selection as a major force structuring group composition. We used 9 microsatellites designed for E. alba to determine the genetic identity and probability of parentage of individuals. We analyzed parentage and kinship using the software ML-Relate, GenAIEx, and Cervus. The obtained relationship probabilities (0.5) revealed a clear maternal relationship between female adults and offspring with allele compatibility, and at least 5 relationships between male adults and pups. We found a low degree of relatedness within roosting groups. Between roosting groups at different sites, the mean probability of a half-sibling relationship ranged from 0.214 to 0.244 and, for full-sibling relationship, from 0.383 to 0.553. Genetically, adult individuals were poorly related within clusters, and kinship as an evolutionary force could not explain group membership.     

Fluorescence of Extracts from Argentine Hake (Merluccius hubbsi) Muscle and Its Correlation with Quality Indices During Chilled Storage

Argentine hake (Merluccius hubbsi) was stored chilled for 8 days. The evolution of fluorescence of the aqueous and organic extracts of muscle during storage at 4°C was studied and correlated with common damage indices; pH, total volatile basic nitrogen, free fatty acids, and the fluorescence intensity of the aqueous phase showed a good correlation with storage time. The excitation-emission matrices of the aqueous extracts were also obtained. A fluorescence landscape that could be attributed to tryptophan decreased with storage time, meanwhile a peak (around 340 and 400 nm for excitation and emission wavelengths, respectively) is developed during chilled storage.     

Morphological development and allometric growth of yellowtail kingfish Seriola lalandi V. larvae under culture conditions

Morphological development and allometric growth patterns of Seriola lalandi larvae were assessed to characterize normal growth patterns under culture conditions. Early ontogenetic stages of yellowtail kingfish exhibited an exponential growth in terms of standard length as a function of age. Five development stages were characterized from hatching to the juvenile stage: larval stage I (0–2 days post hatch, dph) with endogenous feeding, characterized by a small yolk sac, unpigmented eyes, primordial finfold surrounding the body and a closed mouth; larval stage II (2–15 dph) characterized by mouth opening, complete pigmentation of eyes and the beginning of the exogenous feeding; subsequently, in the larval stage III (15–25 dph) the posterior tip of notochord of the larvae bended upward and the first rays appeared in fins, concomitant with a change in swimming behaviour; thereafter, larval stage IV (post‐flexion stage; 25–30 dph) began when larvae resembled in morphology to a juvenile organism; however, caudal and dorsal fins were not completely development. Lastly, the juvenile stage was reached 30 dph characterized by a morphology and fin structures similar to those of the adults. Growth and development of structures and organs associated with vital functions such as feeding, sensorial and breathing systems seemed to be more critical previous to 23 dph, which was reflected with a positive allometric growth of head and eyes during this period. The results from this study can be used as a tool‐guide to assess normal development in larval research with S. lalandi to improve existing rearing protocols in hatchery production.     

Probiotic microorganisms and antiviral plants reduce mortality and prevalence of WSSV in shrimp (Litopenaeus vannamei) cultured under laboratory conditions

The protective effect of a probiotic mixture (PM) and antiviral plants, against the white spot syndrome virus (WSSV) in Litopenaeus vannamei , was evaluated in three experiments. The PM was composed of four lactic acid bacteria (LAB) and one yeast strain. The plant mixture was composed of Ocimum sanctum and commercial antiviral plants (VPH^®, HSV^®). Shrimp in each experiment (weighing 2.7±0.7, 11.5±1.3, 11.70±2.5 g) were cultured in 120-L plastic tanks and fed twice a day with commercial feed plus additives (plants or bacteria and yeast). Animals were monitored for the occurrence of WSSV by single-step and nested PCR. The PM and powdered antiviral plants added to the commercial feed showed an increase in survival and a decrease in the prevalence of WSSV in shrimp. The results showed that both the PM and the powdered antiviral plants can provide protection for shrimp against WSSV.     

Intermediate landscape disturbance maximizes metapopulation density

The viability of metapopulations in fragmented landscapes has become a central theme in conservation biology. Landscape fragmentation is increasingly recognized as a dynamical process: in many situations, the quality of local habitats must be expected to undergo continual changes. Here we assess the implications of such recurrent local disturbances for the equilibrium density of metapopulations. Using a spatially explicit lattice model in which the considered metapopulation as well as the underlying landscape pattern change dynamically, we show that equilibrium metapopulation density is maximized at intermediate frequencies of local landscape disturbance. On both sides around this maximum, the metapopulation may go extinct. We show how the position and shape of the intermediate viability maximum is responding to changes in the landscape’s overall habitat quality and the population’s propensity for local extinction. We interpret our findings in terms of a dual effect of intensified landscape disturbances, which on the one hand exterminate local populations and on the other hand enhance a metapopulation’s capacity for spreading between habitat clusters.     

Comparative intestinal absorption of amino acids in rainbow trout (Oncorhynchus mykiss), totoaba (Totoaba macdonaldi) and Pacific bluefin tuna (Thunnus orientalis)

The kinetics of amino acid absorption in the proximal section of three fish species were studied using an everted intestine technique and a pancreatic digest of casein (Tryptone), as the model amino acid mixture. Fresh intestine of rainbow trout (Oncorhynchus mykiss) was used to set up the experimental system, and results were compared with those obtained using intestines from totoaba (Totoaba macdonaldi) and bluefin tuna fish (Thunnus orientalis). The kinetics of amino acid absorption was sigmoidal for all amino acids and for each species of fish tested. In general, specific absorptions of essential amino acids were higher than those of non‐essential ones. No correlation between the concentration of amino acids in the Tryptone solution and the corresponding absorptions was found. The maximum specific absorption rates of all amino acids for trout were 10 times higher than those determined for totoaba and bluefin tuna. The relative amounts of the different amino acids preferentially absorbed in all three species were different. Results obtained from the everted technique may be applicable to the design of formulated diets for large fish species with commercial value.     

Effect of nutrient solution salinity and ionic concentration on parsley (Petroselinum crispum Mill.) essential oil yield and content

The growth and essential oil (EO) production of parsley were evaluated in response to salinity and nutrient solution concentrations in a soilless culture. Parsley plants that were 60 days old were potted in a coconut fiber and peat moss medium and were treated with four different nutrient solutions, including T1, T2, T3 and T4. The T1 nutrient solution was the standard, the T2 and T3 solutions contained incremental macronutrient concentrations with an electrical conductivity (EC) of up to 2.2 and 3.2 dS m^?1, respectively, and the T4 solution was the same as T2 but with sodium chloride (NaCl) and an incremental macronutrient concentration with an EC of 3.2 dS m^?1. Next, these plants were grown for 90 days in a greenhouse with natural daylight in Nador, Morocco. Shoot and root growth significant decreased with increasing EC. However, the salinity that resulted from the addition of NaCl did not affect plant growth in the nutrient solutions. The optimum obtained growth and EO production were 1.2 and 2.2 dS m^?1, respectively. Consequently, the optimum EC value (based on the EO production) of parsley in the soilless culture was 1.2–2.2 dS m^?1.     

Influence of Salinity in the Bioavailability of Zn in Sediments of the Gulf of Cádiz (Spain)

This study assesses the effect of salinity in bioavailability and toxicity of Zn by means of laboratory bioassays by observing contamination in both sediment and water, accumulation of Zn in biological tissues, and histopathological damage in the gills and guts tissues of Ruditapes philippinarum clams, which were exposed to different types of sediments from the Gulf of Cádiz (SW Spain) as well as two dilutions of toxic mud coming from an accidental mining spill. With this objective, the coefficients of distribution (K _D) for Zn between overlying water and sediments were calculated, the histopathological frequencies in the tissues of the gills and guts of clams were determined, and the biota-sediment bioaccumulation factors as well as the bioaccumulation factors were quantified in the different stations. Results showed that the greatest histopathological damages appeared when the salinity values decreased. Statistical results showed that salinity was inversely correlated with histopathological damage (p?<?0.01) for the lesion index for gills. The most outstanding results were observed in the two dilutions of toxic mud (0.3% and 7.9%) at a salinity value of 10. Salinity was inversely correlated with the concentration of Zn in biological tissues (p?<?0.05) and inversely correlated with the concentration of Zn in water and sediment. Zn mobilization to the overlying water is produced when salinity values decrease.