The temperature dependence of the development of the embryo in Mesocyclops leuckarti (Claus) and Cyclops scutifer

This work is mainly intended as an addition to the studies of the populations dynamics of Cyclops scutifer, which is part of the ”Latn ja jaure project” (a study of the principles involved in the ecosystem of a small -initially fish free- mountain lake, before and after the introduction of fish). The field work consisted of sampling in Lake Erken in Roslagen in June, July and September, as well as in Latn ja jaure in the Abisko mountains in August and September of 1965. Additional sampling was done in Latn ja jaure for the study of the horizontal, vertical and temporal distribution of Cyclops scutifer, as well as the in situ development of the different stages. These samples have been analysed in such a way as to fit into the frame work of future studies on the population dynamics of Cyclops scutifer, The main aim of the present investigation is the determination of the dependence upon tempera- tare of the development of the embryo in the subarctic Cyclops scutifer as compared with the conditions found in the warm water species Mesocyclops leuckarti.

A case of development of the parthenogenetic embryo in the ovary of Moina macrocopa Straus (Crustacea, Cladocera). [Translation from: Vest. Zool., Kiev 4, p. 86, 1971. ]

In a survey of histological preparations of several hundreds of Moina macrocopa Straus , one specimen was discovered, in the left ovary of which was found a parthenogenetic embryo. The short article includes an illustration of a transverse section of parthenogenetic female of Moina macrocopa.

The ontogeny of haematopoiesis in the marine teleost Leiostomus xanthurus and a comparison of the site of initial haematopoiesis with Opsanus tau

The ontogeny of haematopoiesis in the perciform fish, spot Leiostomus xanthurus, differed from that reported as the norm for fishes, as exemplified by the cypriniform zebrafish Danio rerio, and observed in the batrachoidiform oyster toadfish Opsanus tau. Erythropoiesis in spot was first evident in the head kidney of yolk-sac larvae 3 days after hatching (DAH). No embryonic intermediate cell mass (ICM) of primitive stem cells or blood islands on the yolk were apparent within embryos. Erythrocytes were first evident in circulation near the completion of yolk absorption, c. 5 DAH, when larvae were c. 20 mm notochord length (LN). Erythrocyte abundance increased rapidly with larval development for c. 14 to 16 DAH, then became highly variable following changes in cardiac chamber morphology and volume. Erythrocytic haemoglobin (Hb) was not detected within whole larvae until they were 12 DAH or c. 31 mm LN, well after yolk and oil-globule absorption. The Hb was not quantified until larvae were >47 DAH or >7 mm standard length. The delayed appearance of erythrocytes and Hb in spot was similar to that reported for other marine fishes with small embryos and larvae. In oyster toadfish, a marine teleost that exhibits large embryos and larvae, the ICM and Hb were first evident in two bilateral slips of erythropoietic tissue in the embryos, c. 5 days after fertilization. Soon thereafter, erythrocytes were evident in the heart, and peripheral and vitelline circulation. Initial haematopoiesis in oyster toadfish conformed with that described for zebrafish. While the genes that code for the development of haematopoiesis are conserved among vertebrates, gene expression lacks phylogenetic pattern among fishes and appears to conform more closely with phenotypic expression related to physiological and ecological influences of overall body size and environmental oxygen availability.

Survey of sediment quality in Sabine Lake, Texas and vicinity

The toxicity of sediments in Sabine Lake, Texas, and adjoining Intracoastal Waterway canals was determined as part of bioeffects assessment studies managed by NOAA’s National Status and Trends Program. The objectives of the survey were to determine: (1) the incidence and degree of toxicity of sediments throughout the study area; (2) the spatial patterns (or gradients) in chemical contamination and toxicity, if any, throughout the study area; (3) the spatial extent of chemical contamination and toxicity; and (4) the statistical relationships between measures of toxicity and concentrations of chemicals in the sediments. Surficial sediment samples were collected during August, 1995 from 66 randomly-chosen locations. Laboratory toxicity tests were performed as indicators of potential ecotoxicological effects in sediments. A battery of tests was performed to generate information from different phases (components) of the sediments. Tests were selected to represent a range in toxicological endpoints from acute to chronic sublethal responses. Toxicological tests were conducted to measure: reduced survival of adult amphipods exposed to solid-phase sediments; impaired fertilization success and abnormal morphological development in gametes and embryos, respectively, of sea urchins exposed to pore waters; reduced metabolic activity of a marine bioluminescent bacteria exposed to organic solvent extracts; and induction of a cytochrome P-450 reporter gene system in exposures to solvent extracts of the sediments. Chemical analyses were performed on portions of each sample to quantify the concentrations of trace metals, polynuclear aromatic hydrocarbons, and chlorinated organic compounds. Correlation analyses were conducted to determine the relationships between measures of toxicity and concentrations of potentially toxic substances in the samples. Based upon the compilation of results from chemical analyses and toxicity tests, the quality of sediments in Sabine Lake and vicinity did not appear to be severely degraded. Chemical concentrations rarely exceeded effects-based numerical guidelines, suggesting that toxicant-induced effects would not be expected in most areas. None of the samples was highly toxic in acute amphipod survival tests and a minority (23%) of samples were highly toxic in sublethal urchin fertilization tests. Although toxic responses occurred frequently (94% of samples) in urchin embryo development tests performed with 100% pore waters, toxicity diminished markedly in tests done with diluted pore waters. Microbial bioluminescent activity was not reduced to a great degree (no EC50 <0.06 mg/ml) and cytochrome P-450 activity was not highly induced (6 samples exceeded 37.1 ug/g benzo[a]pyrene equivalents) in tests done with organic solvent extracts. Urchin embryological development was highly correlated with concentrations of ammonia and many trace metals. Cytochrome P450 induction was highly correlated with concentrations of a number of classes of organic compounds (including the polynuclear aromatic hydrocarbons and chlorinated compounds). (PDF contains 51 pages)

Der Fischei-Test: Ein Toxizitätstest für ökotoxikologische Untersuchungen

Zebrafish (Danio rerio) embryos have been used to quantify the teratogenic potential of environmental samples and harmful substances respectively. The short spawning interval renders this species a good test organism in toxicological research. Due to the transparency of the eggs several lethal and non-lethal endpoints can be detected in parallel after 48 h of embryonic development. Zebrafishembryos have been shown to be sensitive to a number of environmental relevant contaminants, as well as to ex-tracts from polluted sediments

Embryonic development in Clarias gariepinus under laboratory conditions

The embryonic development in Clarias gariepinus was studied under laboratory conditions. The developmental stages of eggs starting from first cleavage were examined microscopically. Photomicroscope was used to take important stages of segmentation, blastulation, differentiation of embryo and hatching. The films of the photograph were developed and printed for each stage produced. The accurate timing and detailed description of each stage was done. The results show that the blastodisc (Polar cap) appeared about 35 minutes after fertilization and the first cleavage dividing the blastodisc into two blastomeres occurs 15 minutes after polar cap formation. Details of the developmental stages of embryos and the timing from one stage to the other were described. The larva shook off the shell and emerged completely from the egg case about 22 hours after fertilization at a water temperature of 25.1 degree C. The accurate determination of the time of initiation of first mitosis is of great importance in fish culture and breeding especially in the production of tetraploids

Induced diploid meiogynogenesis in the African clariid catfish Heterobranchus longifilis (Valenciennes, 1840) (Pisces: Clariidae)

Diploid meiotic gynogenesis was induced in African catfish, Heterobranchus longifilis by injection of 0.5ml/kg ovaprim on the breeders, followed by application of UV light irradiation on the spermatozoa and temperature shocking of activated eggs. Diploidy was restored by shocking haploid activated eggs at 5 degree C for 40 minutes. The normal control spermatozoa did not receive any UV irradiation nor temperature shock, while the haploid control spermatozoa were irradiated, but did not receive cold shock. The percentage hatchability in the treated group was 25%, while in the control it was 53%. Less than 15 fingerlings had morphological aberrations. After two weeks of indoor rearing, the survival percentage of the treated group was 45% in the control experiment. Cytogenetic analysis of chromosomes revealed 25 chromosomes in the haploid embryo and 50 chromosomes each in diploid gynogenesis and normal diploid control

Triploidy induction in Heterobranchus longifilis (Family: Clariidae) by cold shock

Triploid was induced in African Catfish (Heterobranchus longifilis) by cold shocking activated eggs at 5 degree C for forty minutes starting 3-4 minutes after fertilization. Triploidy was confirmed from mitotic chromosomes prepared from embryo which showed 100% triploidy in the cold shocking treatment and 100% diploidy in the control treatment

Embryogenesis of Heterobranchus longifilis (Curvier and Valenciennes, 1840)

Studies on development of H. longifilis (Curvier and Valenciennes, 1840) were conducted at a temperature of 25EC ( 1Ec) in aquaria tanks continuous development were monitored with the use of wild Heerbrugy photomacroscope and length of yolk and larva were monitored using Stereo Olympus microscope with ocular micrometer. The division into animal and vegetal poles was observed 22 minutes after activation. The first cleavage occurred 65 minutes after activation while the second division which was perpendicular to the first line of division occurred 74 minutes after activation. This was quickly followed by the third and fourth cleavage at 80th and 82nd minutes after activation respectively. Morular stage was reached at 4 hours 20 minutes with formation of optic bud at 14 hours 35 minutes. (DBO) Developing embryo hatched after 27 hours of activation at a mean length of 6.63 and mean yolk length of 2.17. Yolk size decrease at an average rate of 38.5 % till the 5th day of total absorption. Growth of larvae proceeded faster in tail-anus region than in anus-snout portion of the body. The rate of yolk absorption and larva development (survival) as monitored in this work gives important information in Research and development programme for H. longifilis larva - an important aspect of Research development and implementation of appropriate technologies in small scale fisheries

A nursery site of the Alaska skate (Bathyraja parmifera) in the eastern Bering Sea

A nursery site for the Alaska skate (Bathyraja parmifera) was sampled seasonally from June 2004 to July 2005. At the small nursery site (~2 km2), located in a highly productive area near the shelf-slope interface at the head of Bering Canyon in the eastern Bering Sea, reproductive males and females dominated the catch and neonate and juvenile skates were rare. Seasonal samples showed summertime (June and July) as the peak reproductive time in the nursery although some reproduction occurred throughout the year. Timeseries analysis of embryo length frequencies revealed that three cohorts were developing simultaneously and the period of embryonic development was estimated at 3.5 years and average embryo growth rate at 0.2 mm/day. Estimated egg case deposition occurred mainly during summertime and hatching occurred during winter months. Protracted hatching times may be common for oviparous elasmobranch species and may be directly correlated with ambient temperatures as evident from a meta-data analysis. Evidence indicates that the Alaska skate uses the eastern Bering Sea outer continental shelf region for reproduction and the middle and inner shelf regions as habitat for immature and subadults. Skate nurseries may be vulnerable to disturbances because they are located in highly productive areas and because embryos develop slowly.

Exploitation and reproduction of the spotted eagle ray (Aetobatus narinari) in the Los Frailes Archipelago, Venezuela

We studied a small artisanal fishery for the spotted eagle ray (Aetobatus narinari) off Margarita Island in northeastern Venezuela. We analyzed data from 413 fishing trips directed at A. narinari over a 29-month sampling period (August 2005–December 2007). These trips yielded 55.9 metric tons and 1352 individuals from which a subsample of 846 females and 321 males was used for biological data. Maximum fishing effort and landings occurred between February and May, and catch per unit of effort was highest between December and February and between July and October with an overall average of 3 individuals and 133 kg per trip. The overall sex ratio was significantly different from 1:1 with a predominance of females. Females ranged in size with disc widths (DW) from 64 to 226 cm. Males ranged in size between 97 and 190 cm DW. There was no statistically significant difference between male and female length-weight relationships. Mean fecundity was estimated at 3.09 embryos per female, and the largest embryo measured 44.5 cm DW. Females in different maturity stages were found in all months, except November 2007, the month when all females were immature. Postgravid females occurred mainly during the periods of January–May and July–October. Mean length (L50) at maturity was estimated at 129.2 cm DW for males and 134.9 cm DW for females. This study provides much needed information on the biology and life history of A. narinari for the management of an intensive, directed, small-scale fishery for this little known species in northeastern Venezuela.

Maturity, ovarian cycle, fecundity, and age-specific parturition of black rockfish (Sebastes melanops)

From 1995 to 1998, we collected female black rockfish (Sebastes melanops) off Oregon in order to describe their basic reproductive life history and determine age-specific fecundity and temporal patterns in parturition. Female black rockfish had a 50% probability of being mature at 394 mm fork length and 7.5 years-of-age. The proportion of mature fish age 10 or older significantly decreased each year of this study, from 0.511 in 1996 to 0.145 in 1998. Parturition occurred between mid-January and mid-March, and peaked in February. We observed a trend of older females extruding larvae earlier in the spawning season and of younger fish primarily responsible for larval production during the later part of the season. There were differences in absolute fecundity at age between female black rockfish with prefertilization oocytes and female black rockfish with fertilized eggs; fertilized-egg fecundity estimates were considered superior. The likelihood of yolked oocytes reaching the developing embryo stage increased with maternal age. Absolute fecundity estimates (based on fertilized eggs) ranged from 299,302 embryos for a 6-year-old female to 948,152 embryos for a 16-year-old female. Relative fecundity (based on fertilized eggs) increased with age from 374 eggs/g for fish age 6 to 549 eggs/g for fish age 16.

Cloning and expression of zebra fish ferroportin and investigating its influence on anemia

Iron is required for many microbes and pathogens for their survival and proliferation including Leishmania which cause leishmaniasis. Leishmaniasis is an increasingly serious infectious disease with a wide spectrum of clinical manifestations. These range from localized cutaneous leishmaniasis (CL) lesions to a lethal visceral form. Certain strains such as BALB/c mice fail to control L. major infection and develop progressive lesions and systemic disease. These mice are thought to be a model of non-healing forms of the human disease such as kala-azar or diffuse cutaneous leishmaniasis. Progression of disease in BALB/c mice has been associated with the anemia, in last days of their survival, the progressive anemia is considered to be one of the reasons of their death. Ferroportin (Fpn), a key regulator of iron homeostasis is a conserved membrane protein that exports iron across the duodenal enterocytes as well as macrophages and hepatocytes into the blood circulation. Fpn has also critical influence on survival and proliferation of many microorganisms whose growth is dependent upon iron, thus preparation of Fpn is needed to study the role of iron in immune responses and pathogenesis of micoorganisms. To prepare and characterize a recombinant ferroportin, total RNA was extracted from Indian zebrafish duodenum, and used to synthesize cDNA by RT-PCR. PCR product was first cloned in Topo TA vector and then subcloned into the GFP expression vector pEGFP–N1. The final resulted plasmid (pEGFP-ZFpn) was used for expression of FPN-EGFP protein in Hek 293T cells. The expression was confirmed by fluorescence microscopy and flow cytometery. Recombinant Fpn was further characterized by submission of its predicted amino acid sequences to the TMHMM V2.0 prediction server (hidden Markov model), NetOGlyc 3.1 server and NetNGlyc 3.1 server. Data emphasised that obtained Fpn from indian zebrafish contained eight transmembrane domains with N- and C-termini inside the cytoplasm and harboured 78 mucin-type glycosylated amino acid. The results indicate that the prepared and characterized recombinant Fpn protein has no membrane topology difference compared to other Fpn described by other researcher. Our next aim was to deliver recombinant plasmid (pEGFP-ZFpn) to entrocyte cells. However, naked therapeutic genes are rapidly degraded by nucleases, showing poor cellular uptake, nonspecificity to the target cells, and low transfection efficiency. The development of safe and efficient gene carriers is one of the prerequisites for the success of gene therapy. Chitosan and alginate 139 polymers were used for oral gene carrier because of their biodegradability, biocompatibility and their mucoadhesive and permeability-enhancing properties in the gut. Nanoparticles comprising Alginate/Chitosan polymers were prepared by pregel preparation method. The resulting nanoparticles had a loading efficiency of 95% and average size of 188 nm as confirmed by PCS method and SEM images had showed spherical particles. BALB/c mice were divided to three groups. The first and second group were fed with chitosan/alginate nanoparticles containing the pEGFP-ZFpn and pEGFP plasmid, respectively (30 μgr/mice) and the third group (control) didn’t get any nanoparticles. The result showed BALB/c mice infected by L.major, resulted in higher hematocryte and iron level in pEGFP-ZFpn fed mice than that in other groups. Consentration of cytokines determined by ELISA showed lower levels of IL-4 and IL-10 and higher levels of IFN-γ/IL-4 and IFN-γ/IL-10 ratios in pEGFP-ZFpn fed mice than that in other groups. Morover more limited increase of footpad thickness and significant reduction of viable parasites in lymph node was seen in pEGFP-ZFpn fed mice. The results showed the first group exhibited a highr hematocryte and iron compared to the other groups. These data strongly suggests the in vivo administration of chitosan/alginate nanoparticles containing pEGFP-ZFpn suppress Th2 response and may be used to control the leishmaniasis .