Annual reproductive cycle based on histological changes in the ovary of the female mosquitofish,Gambusia affinis, in central Japan

To clarify the annual reproductive cycle of wild female mosquitofish,Gambusia affinis, in Mie Prefecture, central Japan, changes in ovarian histology were investigated. Female mosquitofish kept in aquaria under constant temperature (25°C) and photoperiod (16L: 8D) conditions produced successive broods at intervals of 22.1±0.46 days (n=7). Between days 0–3 following parturition, females began active vitellogenesis. Between days 3–5, fully grown oocytes matured and were fertilized, and embryonic development began in the follicles. By day 10, as fertilized eggs continued embryonic development, some oocytes at the oil-droplet stage had begun to accumulate yolk globules for the next gestation. Thus, vitellogenesis of the succeeding batch of oocytes overlaps with gestation during reproduction in the mosquitofish. A rearing experiment showed the annual reproductive cycle of mosquitofish breeding in Nagashima to be as follows. Although oocytes had not at that point developed to the yolk globule stage, copulation occurred in February. Females began vitellogenesis in early May, the first pregnancy of the year commencing in mid-May. From mid-May to August, females repeated the gestation cycle (vitellogenesis, maturation, fertilization, pregnancy and parturition) at around one month intervals. In September, oocyte recruitment from the oil-droplet to the yolk globule stage ceased. After the final parturition, the ovaries contained only non-vitellogenic oocytes. Spermatozoa in the ovarian cavity were scare from November to January.     

Female masculinization and reproductive life history in the western mosquitofish (<Emphasis Type="Italic">Gambusia affinis</Emphasis>)

In this study, we explored the relationship between female masculinization and various morphological and reproductive parameters of male and female western mosquitofish (Gambusia affinis), including body size, anal fin length, testes mass and female fecundity. We detected high levels of female masculinization in the Brazos and Little Brazos Rivers (Falls County, Texas) in March 2008, but not 2009. In both populations, masculinized females were smaller, possessed shorter anal fins, and had significantly lower total fecundity (collective number of eggs and embryos) compared to non-masculinized females. In addition, 2008 non-masculinized females were smaller and less fecund than 2009 non-masculinized females in both populations. Moreover, 2008 males were smaller, had shorter gonopodia, and smaller testes than 2009 males. Our results support the hypothesis that female masculinization can have a strong influence on reproductive life history and population dynamics of livebearing fishes.     

Attenuation of fibrosis <Emphasis Type=Italic>in vitro</Emphasis> and <Emphasis Type=Italic>in vivo</Emphasis> with <Emphasis Type=Italic>SPARC</Emphasis> siRNA

Introduction  

SPARC is a matricellular protein, which, along with other extracellular matrix components including collagens, is commonly over-expressed in fibrotic diseases. The purpose of this study was to examine whether inhibition of SPARC can regulate collagen expression in vitro and in vivo, and subsequently attenuate fibrotic stimulation by bleomycin in mouse skin and lungs.     

Cryopreservation of <Emphasis Type=Italic>Robinia</Emphasis><Emphasis Type=Italic>pseudoacacia</Emphasis>

Cryopreservation of Robinia pseudoacacia explants by vitrification achieved 78% survival following the stepwise preculture of shoot tips in (0.3 + 0.5 + 0.7 M) sucrose with a 80 min incubation in PVS2; compared to 87% survival after desiccation of explants to 30% water content, following 3 days alginate bead (with glycerol and sucrose treatments) preculture in 0.7 M sucrose.     

Probiotic <Emphasis Type=Italic>Lactobacillus</Emphasis> strains: <Emphasis Type=Italic>in vitro</Emphasis> and <Emphasis Type=Italic>in vivo</Emphasis> studies

Three Lactobacillus strains (LOCK 0900, LOCK 0908, LOCK 0919) out of twenty-four isolates were selected according to their antagonistic activity against pathogenic bacteria, resistance to low pH and milieu of bile salts. Intragastric administration of a mixture of these strains to Balb/c mice affected cytokine T_H1-T_H2 balance toward nonallergic T_H1 response. Spleen cells, isolated from lactobacilli-treated mice and re-stimulated in vitro with the mixture of heat-inactivated tested strains, produced significantly higher amounts of anti-allergic tumor necrosis factor- and interferon-γ than control animals whereas the level of pro-allergic interleukin-5 was significantly lower. Lactobacillus cells did not translocate through the intestinal barrier into blood, liver and spleen; a few Lactobacillus cells found in mesenteric lymph nodes could create antigenic reservoir activating the immune system. The mixture of Lactobacillus LOCK 0900, LOCK 0908 and LOCK 0919 strains represents a probiotic bacterial preparation with possible use in prophylaxis and/or therapy of allergic diseases.     

New insights into the in vitro organogenesis process: the case of <Emphasis Type=Italic>Passiflora</Emphasis>

We present evidences that ultrastructural electron microscope findings are valuable ways to understand the in vitro regeneration process, in particular in the yellow passion fruit. Shoot-regeneration was induced in hypocotyl and leaf-derived explants using 4.44 μM BAP, and the entire organogenic process was analyzed using conventional histology, scanning and transmission electronic microscopy. Both direct and indirect regeneration modes were observed in hypocotyl explants, but only direct regeneration occurred in leaf-derived cultures. In the direct pathway from both explant types, meristemoids developed into globular structures, here called protuberances. The peripheral meristematic layers of the protuberances displayed ultrastructural characteristics indicative of a high metabolic activity, and only these cells originated shoots and leaf primordia, the latter being frequent when leaf explants were used. Moreover, the peripheral cells of the protuberances derived from leaf explants lost adhesion during the culture, diminishing the regeneration rates. We recommend the use of hypocotyls as a source of explant to obtain shoots as well as a genetic transformation system for the yellow passion fruit. However, the direct pathway is preferred because a type of amitosis occurred in the peripheral cells of hypocotyl-derived calli, which has the potential to result in genetic instability of the regenerating plants/tissue.     

Regulation of the <Emphasis Type=Italic>ALBINO3</Emphasis>-mediated transition to flowering in <Emphasis Type=Italic>Arabidopsis</Emphasis> depends on the expression of <Emphasis Type=Italic>CO</Emphasis> and <Emphasis Type=Italic>GA1</Emphasis>

ALBINO3, a homologue of PPF1 in Arabidopsis, encodes a chloroplast protein, and is essential for chloroplast differentiation. In the present study, ALBINO3(−) transgenic plants exhibited a significant decrease in both the number of rosette leaves at bolting and the days before bolting, suggesting the important roles of ALBINO3 in regulating flowering during non-inductive short-day photoperiods. ALBINO3 mRNA was apparently accumulated in shoot apical meristem and floral meristems around the shoot apical meristem in wild-type plants. ALBINO3 might be predominantly involved in inducing the floral repression pathway by activating the expression of TFL1, and by suppressing the expression of LFY, respectively, in the shoot apical meristem. Moreover, the function of ALBINO3 in regulating flowering transition depended on the expression of CO and GA1, because ALBINO3 might function in the downstream integration of the photoperiod-dependent and the photoperiod-independent pathways. These results suggest that ALBINO3 may have an important integrative function in the flowering process in Arabidopsis.     

Abundance estimates of the Indo-Pacific lionfish <Emphasis Type=Italic>Pterois volitans</Emphasis>/<Emphasis Type=Italic>miles</Emphasis> complex in the Western North Atlantic

Less than a decade after being observed off Florida, the invasive Indo-Pacific lionfish is now widely distributed off the southeast coast of the United States. As a step towards measuring invasion impacts to native communities, we examine the magnitude and extent of this invasion by first, compiling reports of lionfish to provide range information and second, estimate lionfish abundance from two separate studies. We also estimate native grouper (epinepheline serranids) abundance to better assess and compare lionfish abundances. In the first study we conducted SCUBA diver visual transect surveys at 17 different locations off the North Carolina coast in water depths of 35–50 m. In the second study, we conducted 27 Remote Operated Vehicle (ROV) transect surveys at five locations from Florida to North Carolina in water depths of 50–100 m. In both studies, lionfish were found to be second in abundance only to scamp (Mycteroperca phenax). Lionfish were found in higher abundance in the shallower North Carolina SCUBA surveys ( ha⁻¹) than in the deep water ROV surveys ( ha⁻¹). Lionfish reports continue to expand most recently into the Bahamas, raising the specter of further spread into the Caribbean and Gulf of Mexico. The potential impacts of lionfish to native communities are likely to be through direct predation, competition and overcrowding. The high number of lionfish present in the ecosystem increases the potential for cascading impacts throughout the food chain. Within the southeast region the combined effects of climate change, overfishing and invasive species may have irreversible consequences to native communities in this region. An erratum to this article can be found at     

Efficient separation and purification of allophycocyanin from <Emphasis Type=Italic>Spirulina</Emphasis> (<Emphasis Type=Italic>Arthrospira</Emphasis>) <Emphasis Type=Italic>platensis</Emphasis>

Allophycocyanin (APC) is a minor component of phycobiliproteins in cyanobacteria and red algae. This paper describes a simple and inexpensive extracting method for isolating APC from Spirulina (Arthrospira) platensis with high efficiency. The crude phycobiliprotein extract was pretreated by ammonium sulfate fractionation. Then, by adding hydroxylapatite into crude phycobiliprotein extract dissolved in 20 mM phosphate buffer (pH 7.0), APC was selectively adsorbed by hydroxylapatite but C-phycocyanin (C-PC) was not. The hydroxylapatite was collected and APC was extracted from the crude phycobiliprotein extract. Then, the enriched APC was washed off from the hydroxylapatite using 100 mM phosphate buffer (pH 7.0). In this simple extracting method it was easy to remove C-PC and isolate APC in large amounts. The absorbance ratio A ₆₅₀/A ₂₈₀ of extracted APC reached 2.0. The recovery yield was 70%, representing 4.61 mg · g⁻¹ wet weight. The extracted APC could be further purified by a simple anion-exchange chromatography with a pH gradient from 5.6 to 4.0. The absorbance ratio A ₆₅₀/A ₂₈₀ of the purified APC reached 5.0, and the overall recovery yield was 43%, representing 2.83 mg · g⁻¹ wet weight. Its purity was confirmed by native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate-PAGE.     

Interaction of <Emphasis Type=Italic>Bdellovibrio bacteriovorus</Emphasis> with bacteria <Emphasis Type=Italic>Campylobacter jejuni</Emphasis> and <Emphasis Type=Italic>Helicobacter pylori</Emphasis>

Interaction of Bdellovibrio bacteriovorus 100NCJB with bacteria Campylobacter jejuni (strains 1, 2, 3, 4, and 5) and Helicobacter pylori, strain TX30a, was confirmed. The results indicate that lytic activity of bdellovibrios both in liquid media and cells attached to a surface was observed. The potential use of the antimicrobial activity of predatory bacteria for environmental bioprotection and public health is discussed.     

Life cycle of <Emphasis Type=Italic>Aylax hypecoi</Emphasis> (<Emphasis Type=Italic>Insecta: Hymenoptera: Cynipidae</Emphasis>), a gall inducer on <Emphasis Type=Italic>Hypecoum</Emphasis> ssp. (<Emphasis Type=Italic>Papaveraceae</Emphasis>)

The life cycle and developmental stages of Aylax hypecoi (Trotter, 1913, Hymenoptera: Cynipidae: Aylacini) were studied in detail. Aylax hypecoi is known to induce galls in fruits of two Hypecoum species — H. imberbe and H. geslini (Papaveraceae) and the larva develops in host plant fruits. The morphology and development of egg, larva and pupa were investigated, which has previously not been done. The shape and size of terminal-instar larvae and associated galls are sex-specific. Overwintering stage, adult emergence and flying periods, and egg productivity were studied also.     

Chytrid infection reduces thoracic beat and heart rate of <Emphasis Type=Italic>Daphnia</Emphasis><Emphasis Type=Italic>pulicaria</Emphasis>

Zooplankters are hosts to numerous endo- and ectoparasites, some of which have dramatic impacts on their hosts. Epizootics on zooplankton are probably more widespread in lake systems than it is currently known, and few studies have explored the direct and indirect importance of parasitism in aquatic food webs. In addition, our understanding of the sublethal effects of parasitic infections on host organisms and populations is limited. We used a novel electro-chemical based technique to measure in the outflow of the feeding current changes in the beat rate of the thoracic appendages in female Daphnia pulicaria. We observed simultaneously the heart rates and compared chytrid infected animals with uninfected gravid and non-gravid ones. We found in uninfected animals a thoracic beat rate of 3.81 ± 018 Hz and a heart rate of 4.67 ± 0.42 Hz. Gravid daphnids had a 14% lower thoracic beat rate (3.27 ± 0.30 Hz) than non-gravid females while the heart rate did not significantly differ (4.48 ± 0.28 Hz). In contrast, infected animals showed a 22% lower thoracic beat rate (2.96 ± 0.47 Hz) and a 36% lower heart rate (2.98 ± 0.5 Hz) when compared with uninfected non-gravid females. We discuss the ways Daphnia are affected by Polycaryum leave infections on the individual and population level.     

<Emphasis Type=Italic>In vitro</Emphasis> organogenesis of <Emphasis Type=Italic>Passiflora alata</Emphasis>

Passiflora alata in vitro organogenesis was studied based on explant type, culture medium composition, and incubation conditions. The results indicated that the morphogenic process occurred more efficiently when hypocotyl segment-derived explants were cultured in media supplemented with cytokinin and AgNO₃ incubated under a 16-h photoperiod. The shoot bud elongation and plant development were obtained by transferring the material to MSM culture medium supplemented with GA₃ and incubated in flasks with vented lids. Histological analyses of the process revealed that the difficulties in obtaining plants could be related to the development of protuberances and leaf primordia structures, which did not contain shoot apical meristem. Roots developed easily by transferring elongated shoots to 1/2 MSM culture medium. Plant acclimatization occurred successfully, and somaclonal variation was not visually detected. The efficiency of this organogenesis protocol will be evaluated for genetic transformation of this species to obtain transgenic plants expressing genes that can influence the resistance to Cowpea aphid borne mosaic virus.     

Distribution of <Emphasis Type=Italic>Hordoindoline</Emphasis> genes in the genus <Emphasis Type=Italic>Hordeum</Emphasis>

Hordoindoline (Hin) genes, which are known to comprise Hina, Hinb-1, and Hinb-2, are associated with grain hardness in barley. However, the interspecific variation in the Hin genes in the genus Hordeum has not been studied in detail. We examined the variation in Hin genes and used it to infer the phylogenetic relationships between the genes found in two H. vulgare subspecies (cultivated barley and H. vulgare subsp. spontaneum) and 10 wild relatives (H. bogdanii, H. brachyantherum, H. bulbosum, H. chilense, H. comosum, H. marinum, H. murinum, H. patagonicum, H. pusillum, and H. roshevitzii). The Hina and Hinb genes of these species were amplified by PCR. We found two Hinb genes in three wild species (H. bogdanii, H. brachyantherum, and H. roshevitzii) and preliminarily named them Hinb-A and Hinb-B. Cluster analysis showed that the 17 Hinb genes present in Hordeum formed two distinct clusters (named A and B). Seven Hinb genes were included in Cluster-A, and 10 Hinb genes were included in Cluster-B. All Hinb-A genes were included in Cluster-A, while all of the Hinb-B genes were included in Cluster-B. In contrast, the Hinb-1 and Hinb-2 genes in H. vulgare were included in Cluster-B. These results suggest that the Hinb genes duplicated during the early stages of diversification in the genus Hordeum. On the other hand, the Hinb-1 and Hinb-2 genes in H. vulgare seem to have been generated by a duplication of the Hinb gene after the split of the lineages leading to H. vulgare and H. bulbosum.     

Time Course of <Emphasis Type=Italic>c-fos</Emphasis>, <Emphasis Type=Italic>vasopressin</Emphasis> and <Emphasis Type=Italic>oxytocin</Emphasis> mRNA Expression in the Hypothalamus Following Long-Term Dehydration

Summary 1. This study presents a time course analysis of the messenger RNA (mRNA) levels of c-fos, vasopressin (VP), and oxytocin (OT) in the paraventricular (PVN) and supraoptic nucleus (SON), following acute and chronic dehydration by water deprivation. 2. Male Wistar rats were separated into five groups: nondehydrated (control group) and dehydrated for 6, 24, 48 and 72 h. Following water deprivation, animals were decapitated, their blood was collected for hematocrit, osmolality, and plasma sodium measurements, and brains were removed for dissection of both PVN and SON. 3. As expected, the hematocrit, osmolality, plasma sodium, and weight loss were increased after water deprivation. In SON, a significant increase in both VP and OT mRNA expression was observed 6 h after dehydration reaching a peak at 24 h and returning to basal levels of expression at 72 h. In the PVN, an increase in both VP and OT mRNA expression occurred 24 h after dehydration. At 72 h the VP and OT mRNA expression levels had decreased but they were still at higher levels than those detected in control animals. 4. These results suggest that SON is the first nucleus to respond to the dehydration stimulus. Additionally, we also observed an increase in c-fos mRNA expression in both PVN and SON 6 h after water deprivation, which progressively decreased 24, 48, and 72 h after the onset of water deprivation. Therefore, it is possible that c-fos may be involved in the modulation of VP and OT genes, regulating the mRNA expression levels on a temporally distinct basis within the PVN and SON.