Reproductive biology of Rioraja agassizi from the coastal southwestern Atlantic ecosystem between northern Uruguay (34°S) and northern Argentina (42°S)

A total of 552 individuals of Rioraja agassizi (257 females and 295 males) were collected by bottom-trawl during research cruises. Sexual dimorphism was observed with females are heavier than males for a given total length (TL). Using logistic regression, it was determined that TL at 50% maturity of males was 475 mm TL and of females 520 mm TL. This estimation agrees with the morphological parameters measured. Although not statistically significant, a peak in reproductive activity was observed in males during late spring and summer. The monthly variation in the gonadosomatic index and oviducal gland width, together with the largest diameter of ovarian follicles, suggests that R. agassizi females have a partially defined annual reproductive cycle with two peaks, one from November (spring) to February (summer) and another in July (winter).     

Effect of spermine on interaction of DNA polymerase α from the loach (Misgurnus fossilis) eggs with DNA

Polyamines (putrescine, spermidine and spermine) cause a marked increase in the activity of the loach Misgurnus fossilis DNA polymerase α on activated (gapped) DNA. The stimulatory effect increases in the order: putrescine, spermidine, spermine. Kinetic analysis shows that spermine does not change the affinity of the polymerase for dTTP, but it decreases the enzyme affinity for DNA. The apparent K_m of the polymerase for activated DNA progressively increases from 14 to 1200 μM (nucleotide), if the concentration of spermine rises up to 2 mM, while V_max reaches a maximum at 0.5 mM spermine and then drops at higher polyamine concentrations. Native calf thymus DNA and especially single-stranded DNA from phage M13 appear to be inhibitors of α-polymerase activity on gapped DNA. Dixon plots suggest simple competitive inhibition of the polymerase activity by single- or double-stranded DNA and absence of cooperativity in the interaction of the polymerase with DNA. Hill-plot analysis is compatible with the interpretation that there is only one DNA binding site on each DNA polymerase α molecule. Spermine, even at low concentrations, decreases sharply the affinity of the enzyme for double-stranded DNA, while the enzyme affinity for single-stranded DNA changes insignificantly. Another result of spermine action is the destabilization of the polymerase-DNA complex. The ratio of the ‘static affinity’ of the enzyme to its ‘kinetic affinity’ decreases 2.2-fold in the presence of 0.5 mM spermine. As a result, the sensitivity of DNA synthesis to 3′-deoxy-3′-aminothymidine 5′-triphosphate and to 1-β-d-arabinofuranosylcytidine 5′-triphosphate decreases in the presence of the polyamine. Both spermine effects, the decrease in the ‘nonproductive binding’ of the polymerase to double-stranded regions in DNA and the destabilization of the polymerase-DNA complex, presumably account for the increase in the activity of the loach α-polymerase on activated DNA.     

A phylogenetic analysis of Momordica (Cucurbitaceae) in India based on karyo-morphology,nuclear DNA content and rDNA ITS1–5.8S–ITS2 sequences

The infrageneric delimitation of Momordica, a medicinally important genus of Cucurbitaceae, is ill-defined until date. Momordica chromosomes are extremely small and are difficult to stain and visualize because of the dense cytoplasmic background. We have conducted karyomorphometric analysis by EMA method in five Indian Momordica species, and the nuclear genome sizes were estimated by flow cytometry for the first time. The somatic chromosome numbers ranged from 2n = 18 to 56 in the species. We have resolved previously disputed chromosome numbers in M. cymbalaria and M. dioica as 2n = 18 (lowest) and 2n = 56, respectively. Chromosome counts in the other species were re-confirmed as 2n = 22 in M. charantia, 2n = 28 in M. cochinchinensis and 2n = 56 in M. subangulata. The largest genome size was recorded in M. cymbalaria (3.74 pg 2C⁻¹), while the smallest size (0.72 pg 2C⁻¹) was detected in M. charantia var. charantia. The nuclear genome sizes were analysed in comparison to chromosome numbers and total chromosome lengths of the species. Karyomorphometric indices showed comparable symmetric karyotypes in the species except in M. cymbalaria having tendency towards asymmetry. The UPGMA phenogram and principle component analysis based on nuclear DNA contents and karyomorphometric parameters demonstrated interspecies differences, intraspecific distinction within M. charantia varieties and highlighted distinction of M. cymbalaria. This study was further supported by the rDNA ITS sequence-based phylogenetic analysis which revealed the monophyletic origin of the Indian members of Momordica and clarified the intraspecies relationship among the studied members. As a whole, the study brought out new insights on species diversification within the genus Momordica in India and would benefit further studies on biosystematics and plant breeding programmes.

     

Spatio-temporal variability of ichthyoplankton and its relationship with oceanographic conditions at the shelf break off Chilean Patagonia (43°S–51°S)

Larval fish composition and abundance have been studied in fjords of south Patagonia. However, the ichthyoplankton of the adjacent shelf and slope waters of the Pacific Ocean, which is an important spawning and nursery ground for demersal fishery species (e.g. austral hake and Patagonian grenadier), remains largely unknown. To compare ichthyoplankton occurrence with environmental conditions in August 2013 and 2014 (austral winter), two bio-oceanographic cruises were carried out between 43° and 51°S. Conductivity, temperature and depth (CTD) data and ichthyoplankton samples were collected. The highest frequency of occurrence (FO) was observed for eggs of lightfish Maurolicus parvipinnis (84 and 91% FO in 2013 and 2014, respectively) and larvae of the myctophid Lampanyctodes hectoris (60 and 94% FO, respectively). Ichthyoplankton composition and abundance varied significantly between years, and richness varied from 10 (2013) to 18 taxa (2014). Only larval L. hectoris showed a significant increase in abundance during the winter of 2014. Water column temperature and salinity were significantly different between years; the water was warmer and saltier in 2013. Canonical Correspondence Analysis detected positive relationships between temperature and salinity with egg abundance and larval distribution of Merluccius australis. The highest abundance of eggs of M. australis was found at ～6–10°C, mainly off south Guafo Island. Additionally, ichthyoplankton from the shelf-break area off south Patagonia showed a wider latitudinal distribution in the winter of 2014 than in 2013.     

A comparison of artificial diet and hybrid sweet corn for the rearing of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) based on life table characteristics

The demographic characteristics of Helicoverpa armigera (Hübner) reared on hybrid sweet corn (Zea mays L. variety saccharata) (hybrid super sweet corn KY bright jean) and on an artificial diet were compared by using the age-stage, two-sex life table. Because the hatch rate of eggs varies with maternal age, age-specific fecundity was calculated based on the numbers of hatched eggs to reveal the biological characteristics of H. armigera accurately. The intrinsic rate of increase (r), finite rate (λ) and mean generation time (T) of H. armigera were 0.0853 d(-1), 1.0890 d(-1), and 46.6 d, respectively, on Z. mays and 0.1015 d(-1), 1.1068 d(-1), and 46.3 d, respectively, on the artificial diet. There were significant differences in the intrinsic rate of increase and finite rate between two treatments. The age-stage life expectancy and reproductive value also were calculated. The relationships among the net reproductive rate, the mean female fecundity, the number of emerged females, and the total number of individuals used in the life table study are consistent with theoretical expectations. We recommend the age-stage, two-sex life table for use in insect demographic studies to incorporate both sexes and the variation in developmental rate among individuals and to obtain accurate population parameters. The artificial diet is more suitable for the mass rearing of H. armigera.     

Biases in determining the diet of jumbo squid <Emphasis Type="Italic">Dosidicus gigas</Emphasis> (D’ Orbigny 1835) (Cephalopoda: Ommastrephidae) off southern-central Chile (34°S–40°S)

The diet of jumbo squid (Dosidicus gigas) off southern-central Chile is described to examine potential biases in the determination of their main prey. Specimens were collected from catches using different fishing gear (jigging, trawl and purse-seine), from July 2003 to January 2004, and from December 2005 to October 2006. The stomach contents were analyzed in terms of frequency of occurrence, number, and weight of prey items and the diet composition was analyzed using Detrended Correspondence Analysis. In the industrial purse-seine fleet for jack mackerel (Trachurus murphyi), the dominant prey of D. gigas was T. murphyi. In the industrial mid-trawl fishery for Patagonian grenadier (Macruronus magellanicus), the dominant species in the diet of D. gigas was M. magellanicus. Similarly, Chilean hake (Merluccius gayi) was the main prey in the diet of D. gigas obtained in the industrial trawl fishery for Chilean hake; and, in both artisanal fisheries (purse-seine for small pelagics and jigging), small pelagic fish and D. gigas were the main prey in the stomach contents of D. gigas. Cannibalism in D. gigas varied between different fleets and probably is related to stress behavior during fishing. The Detrended Correspondence Analysis ordination showed that the main prey in the diet of D. gigas is associated with the target species of the respective fishery. Consequently, biases are associated with fishing gear, leading to an overestimate in the occurrence of the target species in the diet. We recommend analyzing samples from jigging taken at the same time and place where the trawl and purse-seine fleets are operating to avoid this problem, and the application of new tools like stable isotope, heavy metal, and fatty acid signature analyses.     

Theoretical description of halogen bonding – an insight based on the natural orbitals for chemical valence combined with the extended-transition-state method (ETS-NOCV)

In the present study we have characterized the halogen bonding in selected molecules H₃N–ICF₃ (1-NH ₃ ), (PH₃)₂C–ICF₃ (1-CPH ₃ ), C₃H₇Br–(IN₂H₂C₃)₂C₆H₄ (2-Br), H₂–(IN₂H₂C₃)₂C₆H₄ (2-H ₂ ) and Cl–(IC₆F₅)₂C₇H₁₀N₂O₅ (3-Cl), containing from one halogen bond (1-NH ₃ , 1-CPH ₃ ) up to four connections in 3-Cl (the two Cl–HN and two Cl–I), based on recently proposed ETS-NOCV analysis. It was found based on the NOCV-deformation density components that the halogen bonding C–X ^… B (X-halogen atom, B-Lewis base), contains a large degree of covalent contribution (the charge transfer to X ^… B inter-atomic region) supported further by the electron donation from base atom B to the empty σ*(C–X) orbital. Such charge transfers can be of similar importance compared to the electrostatic stabilization. Further, the covalent part of halogen bonding is due to the presence of σ-hole at outer part of halogen atom (X). ETS-NOCV approach allowed to visualize formation of the σ-hole at iodine atom of CF₃I molecule. It has also been demonstrated that strongly electrophilic halogen bond donor, [C₆H₄(C₃H₂N₂I)₂][OTf]₂, can activate chemically inert isopropyl bromide (2-Br) moiety via formation of Br–I bonding and bind the hydrogen molecule (2-H ₂ ). Finally, ETS-NOCV analysis performed for 3-Cl leads to the conclusion that, in terms of the orbital-interaction component, the strength of halogen (Cl–I) bond is roughly three times more important than the hydrogen bonding (Cl–HN).

Quality assessment of the predatory bugs Podisus maculiventris (Say) and Podisus sagitta (Fab.) (Heteroptera: Pentatomidae) after prolonged rearing on a meat—based artificial diet

The predatory stinkbugs Podisus maculiventris and Podisus sagitta were reared for more than 15 consecutive generations on a meat—based artificial diet. When they were returned to a diet of live prey (larvae of the pyralid Galleria mellonella) after different generations on the artificial diet, developmental and reproductive traits were similar to those of bugs continuously reared on live prey. Under laboratory conditions, predation rates for nymphs and adults of both pentatomids on larvae of the noctuid Spodoptera exigua were found to be unaffected by the previous diet. The results suggest that long—term rearing of P. maculiventris and P. sagitta on the meat—based artificial diet did not substantially affect the quality of the predators.     

Community structure and food web based on stable isotopes (δN and δC) analysis of a North Eastern Atlantic maerl bed

Maerl beds are highly biodiverse biogenic substrata that have been receiving increasing attention in the last decade. Although maerl beds represent important nursery areas for commercial fishes and molluscs, little is known on the trophic web of their communities. Community structure parameters of maerl bed of the Bay of Brest (species richness, abundance, biomass and dominating species) were studied in parallel with the carbon and nitrogen isotopic composition of their main benthic species (macrofaunal, and megafaunal organisms) in order to assess the trophic levels and differences in the potential food sources of maerl inhabitants. The major potential sources of energy were identified to originate either from epiphytic macroalgae and microphytobenthos both growing on maerl thalli, together with sedimenting (sedimentary) particulate organic matter (POM) originating from the water column. The majority of the macro- and megafaunal organisms investigated were filter feeders, selective-deposit feeders and predators/scavengers. Filter feeders fall into three different groups representing different trophic pathways (i) sponges feeding directly on POM (water column filter feeders I), (ii) ascidians and holothurians feeding on POM and probably captured pelagic preys (water column filter feeders II), and (iii) filter feeding molluscs and crustaceans were hypothesised to feed on microphytobenthos or on decaying sedimented POM (Interface filter feeders). Selective deposit feeders were also divided into two subgroups. Carnivores were also distinguished between those with scavenging habits and true predators. Coupling of the trophic levels observed with the community biomass structure revealed that most of the benthic biomass derives its food from detritic sedimented POM and/or microphytobenthos, with interface filter feeders (23% of the biomass), selective deposit feeders (12%). Carnivores made up to 14% of the total biomass. Generally stable isotopes ratio mean values overlap and cover a large range within feeding types, indicating a strong overlap in food sources and a high degree of complexity of the food web presumably due to the diversity of the potential food sources.     

A new genus and eight new species of the subtribe Anillina (Carabidae,Trechinae, Bembidiini) from Mexico,with a cladistic analysis and some notes on?the?evolution of the genus

One new genus and eight new species of anilline carabids are described from southern Mexico. The new genus, Zapotecanillus gen. n., is established for Z. oaxacanus (type species) sp. n., Z. nanus sp. n., Z. iviei sp. n., Z. ixtlanus sp. n., Z. montanus sp. n., and Z. kavanaughi sp. n. from the Sierra Madre de Oaxaca, Z. pecki sp. n. from the Sierra Madre del Sur, and Z. longinoi sp. n. from the Sierra Madre de Chiapas. A taxonomic key for all described species of Zapotecanillus and a cladistic analysis, based on morphological data, are provided. Morphological, behavioral and biogeographical aspects of the speciation in the genus obtained from the resulting cladogram are discussed.     

CoMFA analysis of tgDHFR and rlDHFR based on antifolates with 6–5 fused ring system using the all-orientation search (AOS) routine and a modified cross-validated r2-guided region selection (q2-GRS) routine and its initial application

We report the development of CoMFA analysis models that correlate the 3D chemical structures of 80 compounds with 6–5 fused ring system synthesized in our laboratory and their inhibitory potencies against tgDHFR and rlDHFR. In addition to conventional CoMFA analysis, we used two routines available in the literature aimed at the optimization of CoMFA: all-orientation search (AOS) and cross-validated r²-guided region selection (q²-GRS) to further optimize the models. During this process, we identified a problem associated with q²-GRS routine and modified using two strategies. Thus, for the inhibitory activity against each enzyme (tgDHFR and rlDHFR), five CoMFA models were developed using the conventional CoMFA, AOS optimized CoMFA, the original q²-GRS optimized CoMFA and the modified q²-GRS optimized CoMFA using the first and the second strategy. In this study, we demonstrate that the modified q²-GRS routines are superior to the original routine. On the basis of the steric contour maps of the models, we designed four new compounds in the 2,4-diamino-5-methyl-6-phenylsulfanyl-substituted pyrrolo[2,3-d]pyrimidine series. As predicted, the new compounds were potent and selective inhibitors of tgDHFR. One of them, 2,4-diamino-5-methyl-6-(2′,6′-dimethylphenylthio)pyrrolo[2,3-d]pyrimidine, is the first 6–5 fused ring system compound with nanomolar tgDHFR inhibitory activity. The HCl salt of this compound was also prepared to increase solubility. Both forms of the drug were tested in vivo in a Toxoplasma gondii infection mouse model. The results indicate that both forms were active with the HCl salt significantly more potent than the free base.     

Asymmetric reduction of (4S)-(+)-carvone catalyzed by baker's yeast: A green method for monitoring the conversion based on liquid–liquid–liquid microextraction with polypropylene hollow fiber membranes

The α,β-unsaturated carbonyl compound (4S)-(+)-carvone was selectively reduced to (1R,2R,4S)-iso-dihydrocarveol using baker's yeasts. The conversion of the bioreduction reaction was monitored using a green hollow-fiber liquid–liquid–liquid microextraction (HF-LLLME) technique. Several parameters which may affect the bioreduction of (4S)-(+)-carvone, such as temperature, time, substrate/enzyme ratio, pH and buffer concentration, were evaluated. The effect of some additives, such as trehalose, DMSO and the ionic liquid [BMIm][PF₆], was also studied. The (1R,2R,4S)-iso-dihydrocarveol was recovered with 52.7% conversion and diastereoisomeric excess >99% after 48 h of reaction at 40 °C in an aqueous monophasic system, with 0.1 mol L^?1 buffer concentration (pH 7.5) and a substrate/yeast cell mass ratio of 8.0 mg g^?1. The HF-LLLME microextraction technique allowed the optimization of the reaction with a reduction of over 99.5% in relation to the use of organic solvents.