Histology and hormonal effect of Ghrelin on ovary of Benni (Barbus sharpeyi) in order to study sexual maturity, zygote and larval generation

Benni (Barbus sharpeyi) is valuable fish that Khuzastan fisheries office propagated it artificially in Susangerd Fish Propagation Center every year. Pituitary gland is used for this aim but female fish lost their fertilization power after 2-3 years, so in present research, new hormone, that is called Ghrelin. The aims of this research are histology, hormonal, zygote and larval generation studies and comparing the results with each other. Ghrelin is a multifunctional peptidyl hormone which increases GTH-II in fish, amphibian, and birds and mammalian so its effect on Benni sexual maturation was studied. Human Ghrelin (hGRL) was obtained from ANASPEC, Canada, with 28 amino acids. In the present study, three levels of ghrelin including 0 (sham treatments), 0.10 (treatment 1) and 0.15 μg/g (treatment 2) body wt and one level of pituitary gland 4000 μg/g (pituitary treatment) with two replications were used. 56 specimens were injected intraperitonealy and their ghrelin level was evaluated immediately after injection and after 24 h. Control fish(n=16) were just injected by physiological saline. For hormonal studies sham and experimental fish(n=40) were anesthetized with MS-222 at a concentration of 250 mg l-1, and blood samples were collected and kept at 4ْC, then spun to collect serum. Serum samples were stores at -20ْC until the RIA for CTH-II. For histology studies immediately after injection a piece of ovary was collected from control fish (Sham zero) after being anesthetized. The sampled ovaries were fixed in Buin solution and embedded in paraffin, and stained to Sections of 5–6 μm using haematoxylin and eosin. The ovarian samples were performed with a compound microscope. Histology and micrometry studies had done. The mature oocytes had given from mature fish, then weighted and the working fecundity were counted. The mature oocytes fertilized, the eggs were incubated and the percentage of fertilization was calculated. After 72h the eggs hatched and the percentage of hatch was counted. The percentage of hindrance was calculated after 6 days. Hormonal results indicate that ghrelin and pituitary increase significantly the GTH-II level in comparison to sham. Macroscopic observations (before taking ovary) showed that ovaries with green colored have couple oval structure located in the abdominal cavity. Microscopic studies of dissected ovaries indicated simultaneous growth of 127 oocytes with 6 stages. The type of the ovary is asynchronous. The results indicated that both of the ghrelin treatment increased the percentage of mature follicles followed by decrease of immature follicles. There were significant differences (P<0.05) between the number of mature and immature follicles. Average diameter of follicle in both of the ghrelin treatment was significantly (P<0.05) declined in the stages of the vitellogenesis when the result compared to the other treatment. Just treatment 1 and pituitary treatment can give mature oocytes. The fecundity of pituitary treatment significantly increase in comparision to ghrelin treatment (P<0.05). In food-restricted fish where endogenous ghrelin levels are known to be increased, a chronic administration of ghrelin induces overt negative effect in releasing mature oocytes. The percentage of fertilization was significantly increase (P<0.05) in ghrelin t. in comparison to pituitary t. and the percentage of hatch was significantly increase (P<0.05) in pituitary t. in comparison to ghrelin t. There was no significant difference (P>0.05) in terms of percentage of hindrance between treatments. In conclusion, the present study demonstrated that ghrelin has positive effect on the level of GTH-II, oocyte maturation, ovarian vitellogenesis and the number of mature follicles of Barbus sharpeyi ovary. Increasing of the mature follicles number reduces their average diameter, indicating stimulating effect of ghrelin in sexual maturation of Barbus sharpeyi.The ghrelin and pituitary treatment have equal chance in the post-stage of spawning.

The effect of spirulina (fresh and dry) on some biological factors in Penaeus semisulcatus larvae

Spirulina is a filamentous cyanobacteria with many applications in food and drug industries, as a food in human, aquaculture, vet and poultry industries… . Semi and mass culture of Spirulina carries out in different countries. This study was carried out in five phases in order to produce this microalga in Iran. The first phase, Spirulina pure stock was imported from Indonesia. After identification of species, it was cultured in laboratory until we took 20 liters of biomass. The semi-mass culture was carried out in green house. Cell concentration and size of Spirulina were recorded during culture daily and their growth rates were calculated. After two weeks, when the size of Spirulina was suitable, biomass of Spirulina was harvested then accumulated Spirulina weighted and dried in 24 hours in laboratory. In order to microbiological study, the samples of Spirulina (dry and fresh) were cultured on blood agar medium and coliforms were counted. The chemical composition of produced Spirulina was measured by standard methods. Fatty acid and amino acid profiles were acquired by GC and HPLC instruments, respectively. The amount of chlorophyll in Spirulina was determined by spectroscopy method. Also astaxanthin pigment as an important carotenoid was measured by HPLC in Spirulina and Penaeus semisulcatus larvae fed on Spirulina. At final phase of this project, larva fed on produced Spirulina (biomass and dry powder) was compared to Z plus, microencapsulated Spirulina (M.C.F) and Chaetoceros algae as control. This experiment was carried from zoa to early post larvae stage then survival and growth rate of larvae were recorded. The growth rate of larvae was evaluated with ANOVA test and survival rate of treatments was assessed by Log Rank (Mantel –Cox) test. Also during larvae stage, two parameters of water such as nitrate and nitrite were measured in zoa, mysis and post larvae stages. The results of this study were shown that colifom counts were 1.85×106 and 92.3×105 coliform per ml in fresh and dry spirulina, respectively. Protein percent of dry spirulina was 50.93 % (dry weight) and the amount of astaxanthin in spirulina and larvae fed on spirulina were 0.21 and 0.01 mg/kg, respectively. The most survival rate of larvae were observed in zoa III (88.8%) with Z plus supplement treatment, in mysis III (76.5%) combination of Z plus and dry spirulina in comparative between treatments. Larvae growth (4.5mm) of control in early post larvae was the best.

Investigation on the species diversity of Pleuronectiformes order in the Iranian coastline water of Persian Gulf

In Iranian Coastline water of Persian Gulf we researched about flat fishes and revision their Scientitic names, Investigated diversity species (Pleuronectiformes) around three provinces such as khouzestan, Bushehr and Hormozgen. The aim focuse on introducting major family and Species Identification for this Purpose 1311 Specimens collected. Then 36 investigation Parameters includes 18 Morphometric, 11 meristic, 7 distribution has been measured in Excel. The descriptive statistic in spss software would be able us to graph dendrograms by Clustering methods, then Design a standerd table and comparised all the datas with it six major family has been found in Iranian Coasline which named: Soleidae, Bothidae Paralichtidae, Cynoglossidae, Psettodidae, Citharidae. Some of the scientific species names such as Pseudorombus arsius, P. annulatus, P. elevatus, P.malayanus, P. triocellatus from Bothidae family Join it to Paralichtidae family and Eurylossa orientalis change It's name from Soleidae family to Brachirus onentalis. Cynoglossus, bilineatus, C.Puncticepts, C.durbaensis, Clachneri, one of the most Important Point that viewed in our samples, founding a new species (C.Sp) Which we couldn't identify it with all the known keys, then sent it to international Scientific references for this porpose. (Family Cynoglossidae); Pseudorombus annulatus, P. elevatus P.arsius, P.malayanus, paralichtodes algoensis, Poecilopstei Javanicus (Family paralichthidae) Arnoglossus aspilos,A.arabicits, Engyprosopon grandisquama, L.pectoralis, Pseudorombus navalensis, Psettina brevirictis (family Bothidae); Psettodes erumei (Family Psettodidae); Citharoides macrolepis (Family Citharidae) in khouzestan Brachirus orientalis, Parachirus marmoratus, solea elongata, Zebrias synapturides (Family Soleidae) Cynoglossus bilineatus, C.puncticeps, C.arel, C.kopsii, C. Capenis (Family cynoglossida); Pseudorombus arsius, P. elevatus, P. malayanus, Poecilopstei javanicus (Family Paralichthidae); Laeops guentheri, Arnoglossus aspilos, Engyprosopon grandisquama (Family Bothidae); Psettodes erumei (Family psettodidae) in Bushehr. Brachirus orientalis, Parachirus marmoratus (Family Soleidae); Cynoglossus arel, C.bilineatus, C.Puncticeps, C. lachheri (Family cynoglossidae); Pseudorombus arisus, P. elevatus, P. malayanus, Poecilopestie j avanicus (Family parlichthidae); Arnoylossus aspilos, A. arabicus, Laeopes guentheri (Family Bothidae); Psettodes erumei (Family Psettodidae); Citharoides macrolepis (Family Citharidae) In Hormozgan. Khozestan has the maximus diversity speices (26 species) and the minimum is Hormozgan (15 Speices). 13 species of five family has never reported inpersian Gulf but we identify them in our studies for the first time. We abserved 29 Species of six family in our studies, comparision between are resualts with other researches shows that this use is the most compelet study in Iranian coastline water of Persian Gulf.

Examining the possibilty of multiplication and breeding of (Schizothorav [sic] zarudnyi) in order to restoration of regional aquatics reservations of Hamoon Lake and taking its advantag [sic] for aquaculture

Schizothorax zarudnvi, is an endemic fish of east country waters. (Triple lagoons of Hamoon and relevant water resources) that in the world it is reported in this resource specially. This fish named Hamoon mahi is one of the most economically valuable species in this region. Because of the recent years droughts, Hamoon logoon has been drive since 2000. Also, semi-wells (a semi natural resource) were affected drastically by recent drought years and their volume reduced to nearly one third of their real volume and resulted in changing at growth and reproduction physiology process in Schizothorax zanidnyi, brood stocks. Beginning of this project was done from October 2003. It's field studies begun (brood catching) since November 2001 by two methods including entangling gairs and at semi wells of Sistan that (Beach seine) had maximum rate of preparing qualified brood stocks. Broods transferred to Cyprinidea reproduction work shop of Zahak and after taking primary measures they stored in to the edaphic pools. Increasing the success safety factor (coefficient) for artificial reproduction of Sthizothorax zarudnyi , identifying the appropriate tune for Hormonal acceptance (physiological preparation of broods) is needed , so this important work was done regularly by histological studies and GSI measurements since November. Highest GSI rates of females (%80.51) and highest IV stage abundance of sexual maturity (%l 00) were observed an march. On the base of this date, Hormone therapy was done on broods on march. The used hormones are as follows Hypophysis. extraction, GnRHa and Anti Dopamin at the dozes of 3-6 ml, 20-30kg and 10-15 ml per kg body weight respectively and 2-3 times from 11-12-80 they were injected. Injected broods kept in to two circumstances, flow-through (rounded pool) and stagnant systems. In stagnant system 14 and 19 individuals of female and male (Schizothorax zauiulnri) broods, respectively injected in 11th, 15111, 19th, and 24th of march 1380. Non of the injected broods in 11 and 15 and 19th march (in stagnant Condition) answered to Hormone therapy. After final injection broods had general less activity and a few of them died. Mean temperature of brood pond waters (daily) which were injected. Fluctuated between 10-25-13. 63°c but injected broods on 24th march had different characteristics. They had pale color and had few fecundity. In this stage of injection they hadn't any successful vulation. After injection, Mean daily water temperature was 15, 88-17, 54°c. In Flowing system, 13-16 individual of males and females respectively were injected on 15th, 19th, 22th and 23th march. None of injected producers on 15th and 19th march with mean daily water temperature of 10, 25-12°c were prepared for spawning but injected producers on 22nd an 23th march with mean daily water temperature of 13.5-1 rc responded about 75-100 percent. (Schizothorax zarudnyi) brood stocks were prepared for spawning after 353-428 hours/day from final injection. Diameter of obtained eggs (before fertilization) was between 1.9-2.3 min and of fertilized eggs was 3.8mm. Fertilized eggs of (Schizothorax zarudnyi) were hatched after 6-7 days with mean water temperature of 17.08°c. Mean length of on one day larvae was 9.47 mm. Larvae was 9.47 mm. Larvae adsorbed the whole yolk sac after , 5-6 days at 17- 1°c and were prepared for releasing in to edaphic pools. Because of the lack of necessary and complementary facilities in the region , they had to release them in to veniros and growing them for 8 days. At the end of 18th day , 35000 larvae (at first) released into an edaphic pond with a volume of 150m2. After growing them for one moth , mean length and weight of new hatched larvae was 29.41 mm and 1.12►r , respectively. With respect to results of this investigation , artificial reproduction of (Schizothorax zarudnyi) Can be possible at 14-17°C and flowing water with Hormonal treatment. It -s breeding has increased development than other cultural specious in the region. Due to high economical value of this specious in Sistan and ti-s specialization east waters of Iran and having high resistance and proper growth There is a need of it's development and reproduction and culture in fish culture fanns (edaphic ponds• two-purpose pools) at the region and country.

Isolatin [sic] the bacteria produced biactive [sic] metabolits [sic] cytotoxic compound in order to prduction [sic] anticancer substance in the coral regin [sic] the Persian Gulf (Larak Island)

In the present investigation the marine bacteria isolated from corals, sponges sea water and sediments of coral regions in the larak Island located in the Persian Gulf and were examined for ability to produce cytotoxic metabolits in order to use as an anticancer compounds. Cytotoxic effect were isolated bacteria from different samples and were examined by Artemia Cytotoxic Bioassay test, in which 4.5 percent of sea waters, 12 percent of sediments and 28 percent of marine invertebrat showed cytotoxic activity, using Brine Shrimp Bioassay test. Streptomyces S-2004 isolated from soft coral specified as Sinularia erecta had LC50=0.5mg/m1 in Brine Shrimp Bioaassay test. The streptomyces S-2004 produced cytotoxic metabolits in low nutrient condition and sea water medium after 7 days on 250 rpm shaken in vitro condition. The extract partially were semipurified. Then ethyl acetate extraction from aceton extracted of bacterial plate had cytotoxic effect (LC50=4.19ktg/m1) in Human epidermoid carcinoma of mouth cells (KB) by using neutral red assay. Morphological effects of this extract on KB cells showed turgescence, cellular blebs and apoptosis which was a proof for anticancer compounds of the extract. It is seems that streptomyces S-2004 is a new strain and could be introduced as a talented bacteria, which produced cytotoxic metabolits.

A critical reassessment of the role of mitochondria in tumorigenesis

Background Mitochondrial DNA (mtDNA) is being analyzed by an increasing number of laboratories in order to investigate its potential role as an active marker of tumorigenesis in various types of cancer. Here we question the conclusions drawn in most of these investigations, especially those published in high-rank cancer research journals, under the evidence that a significant number of these medical mtDNA studies are based on obviously flawed sequencing results. Methods and Findings In our analyses, we take a phylogenetic approach and employ thorough database searches, which together have proven successful for detecting erroneous sequences in the fields of human population genetics and forensics. Apart from conceptual problems concerning the interpretation of mtDNA variation in tumorigenesis, in most cases, blocks of seemingly somatic mutations clearly point to contamination or sample mix-up and, therefore, have nothing to do with tumorigenesis. Conclusion The role of mitochondria in tumorigenesis remains unclarified. Our findings of laboratory errors in many contributions would represent only the tip of the iceberg since most published studies do not provide the raw sequence data for inspection, thus hindering a posteriori evaluation of the results. There is no precedent for such a concatenation of errors and misconceptions affecting a whole subfield of medical research.