Variety of antimicrobial peptides in the Bombina maxima toad and evidence of their rapid diversification

Antimicrobial peptides secreted by the skin of many amphibians play an important role in innate immunity. From two skin cDNA libraries of two individuals of the Chinese red belly toad (Bombina maxima), we identified 56 different antimicrobial peptide cDNA sequences, each of which encodes a precursor peptide that can give rise to two kinds of antimicrobial peptides, maximin and maximin H. Among these cDNA, we found that the mean number of nucleotide substitution per non-synonymous site in both the maximin and maximin H domains significantly exceed the mean number of nucleotide substitution per synonymous site, whereas the same pattern was not observed in other structural regions, such as the signal and propiece peptide regions, suggesting that these antimicrobial peptide genes have been experiencing rapid diversification driven by Darwinian selection. We cloned and sequenced seven genes amplified from skin or liver genomic DNA. These genes have three exons and share the same gene structure, in which both maximin and maximin H are encoded by the third exon. This suggests that alternative splicing and somatic recombination are less likely to play a role in creating the diversity of maximins and maximin Hs. The gene trees based on different domain regions revealed that domain shuffling or gene conversion among these genes might have happened frequently.

Differential expression of the Brunol/CELF family genes during Xenopus laevis early development

The BRUNOL/CELF family of RNA-binding proteins plays important roles in post-transcriptional regulation and has been implicated in several developmental processes. In this study, we describe the cloning and expression patterns of five Brunol genes in Xenopus laevis. Among them, only Brunol2 is maternally expressed and the zygotic expression of the other four Brunol genes starts at different developmental stages. During Xenopus development, Brunol1, 4-5 are exclusively expressed in the nervous system including domains in the brain, spinal cord, optic and otic vesicles. Brunol2 and 3 are expressed in both the somatic mesoderm and the nervous system. Brunol2 is also extensively expressed in the lens. In transfected Hela cells, BRUNOL1, 2 and 3 proteins are localized in both the cytoplasm and the nucleus, while BRUNOL4 and 5 are only present in the cytoplasm, indicating their different functions.

Food and feeding habits of Saurida Tumbil (Block, 1795) off Mumbai waters

The present investigation suggests that Saurida tumbil (Bloch, 1795) is carnivorous and predacious feeding mainly on fishes followed by crustaceans and molluscs. Cannibalism has been observed in the species. Young ones of this species were observed to feed on small fish and Acetes spp., while adults feed on a variety of food. However, fish seems to be the preferred food item. Monthly gastro-somatic index in males indicates poor feeding in the month of february and maximum during august and september. In females, monthly gastro-somatic index indicate poor feeding in december and maximum during january and march. Sex wise study of feeding intensity did not reveal significant difference in males and females.

Fat and water contents of the muscle and gonad of Otolithus argenteus in relation to gonad growth

Fat and water contents of the muscles and gonads in relation to gonad growth in Otolithus argentes from Karwar are reported. Water content showed an inverse relationship to lipid in both the sexes. Variation in somatic body weight was found associated with the fluctuations in lipid contents.

Microsatellite DNA analysis proves nucleus of interspecies reconstructed blastocyst coming from that of donor giant panda

A method for DNA isolation from early development of blastocyst and further analysis of nuclear and mitochondrial DNA was developed in present study. Total DNA was prepared from interspecies reconstructed blastocyst and a giant panda specific microsatellite locus g(010) was successfully amplified. DNA sequencing of the PCR product showed that two sequences of reconstructed blastocysts are the same as that of positive control giant panda. Our results prove that the nucleus of interspecies reconstructed blastocyst comes from somatic nucleus of donor giant panda.

Interspecies implantation and mitochondria fate of panda-rabbit cloned embryos

Somatic cell nuclei of giant pandas can dedifferentiate in enucleated rabbit ooplasm, and the reconstructed eggs can develop to blastocysts. In order to observe whether these interspecies cloned embryos can implant in the uterus of an animal other than th

Phantom mutation hotspots in human mitochondrial DNA

Phantom mutations are systematic artifacts generated in the course of the sequencing process. Contra common belief these artificial mutations are nearly ubiquitous in sequencing results, albeit at frequencies that may vary dramatically. The amount of artifacts depends not only on the sort of automated sequencer and sequencing chemistry employed, but also on other lab-specific factors. An experimental study executed on four samples under various combinations of sequencing conditions revealed a number of phantom mutations occurring at the same sites of mitochondrial DNA (mtDNA) repeatedly. To confirm these and identify further hotspots for artifacts, > 5000 mtDNA electropherograms were screened for artificial patterns. Further, > 30000 published hypervariable segment 1 sequences were compared at potential hotspots for phantom mutations, especially for variation at positions 16085 and 16197. Resequencing of several samples confirmed the artificial nature of these and other polymorphisms in the original publications. Single-strand sequencing, as typically executed in medical and anthropological studies, is thus highly vulnerable to this kind of artifacts. In particular, phantom mutation hotspots could easily lead to misidentification of somatic mutations and to misinterpretations in all kinds of clinical mtDNA studies.

The fecundity of the false trevally, Lactarius lactarius (Bloch and Schneider) along the Karwar coast

A clear knowledge of the reproductive potential or fecundity of a fish is quite an essential pre-requisite for the proper management and conservation of the resources. The fecundity studies are also undertaken to determine the index of diversity dependent factor affecting the population size (Simpson, 1951). Qasim & Qayyam (1963) have detailed the various pathways by which an understanding of fecundity could be used for fishery biological work. The ability of egg production varies within the individual limits such as length, somatic weight, gonadal weight, volume of fish etc.

Epigenetic marks in cloned rhesus monkey embryos: Comparison with counterparts produced in vitro

Until now, no primate animals have been successfully cloned to birth with somatic cell nuclear transfer (SCNT) procedures, and little is known about the molecular events that occurred in the reconstructed embryos during preimplantation development. In man

HSPC117 deficiency in cloned embryos causes placental abnormality and fetal death

Somatic cell nuclear transfer (SCNT) has been successfully used in many species to produce live cloned offspring, albeit with low efficiency. The low frequency of successful development has usually been ascribed to incomplete or inappropriate reprogramming of the transferred nuclear genome. Elucidating the genetic differences between normal fertilized and cloned embryos is key to understand the low efficiency of SCNT. Here, we show that expression of HSPC117, which encodes a hypothetical protein of unknown function, was absent or very low in cloned mouse blastocysts. To investigate the role of HSPC117 in embryo development, we knocked-down this gene in normal fertilized embryos using RNA interference. We assessed the post-implantation survival of HSPC117 knock-down embryos at 3 stages: E9 (prior to placenta formation); E12 (after the placenta was fully functional) and E19 (post-natal). Our results show that, although siRNA-treated in vivo fertilized/produced (IVP) embryos could develop to the blastocyst stage and implanted without any difference from control embryos, the knock-down embryos showed substantial fetal death, accompanied by placental blood clotting, at E12. Furthermore, comparison of HSPC117 expression in placentas of nuclear transfer (NT), intracytoplasmic sperm injection (ICSI) and IVP embryos confirmed that HSPC117 deficiency correlates well with failures in embryo development: all NT embryos with a fetus, as well as IVP and ICSI embryos, had normal placental HSPC117 expression while those NT embryos showing reduced or no expression of HSPC117 failed to form a fetus. In conclusion, we show that HSPC117 is an important gene for post-implantation development of embryos, and that HSPC117 deficiency leads to fetal abnormalities after implantation, especially following placental formation. We suggest that defects in HSPC117 expression may be an important contributing factor to loss of cloned NT embryos in vivo. (C) 2010 Elsevier Inc. All rights reserved.

The study of biological characteristics and heavy metal determination of Cd, Pb, Fe, V, Ni in the muscle, liver and gonad tissues of kingfish Scomberomorus commerson in the coastal waters of the Hormozagan Province (Persian Gulf)

Biological studies and heavy metal (Ni, v, Fe, Pb,Cd) determination in liver, gonad and muscle of Scomberomorus commerson were carried out from Oct 2006 to Sept 2007 in Hormozgan coastal waters. 599 Samples were gutted for reproduction and nutrition studies, fork length and weight were measured to nearest cm and g respectively. Meanwhile 40 samples were also investigated for heavy metal studies. All specimens were collected from two major landing sites (Bandar Lengeh & Bandar Abbas). Minimums & maximum fork length & weight were 29, 128 cm & 235 and 15350 g respectively.Isometric growth was shown according to our study and b was estimated 2.9 (overall), 2.91(male) & 2.89 (female). The average relative gut length was 0.52± 0.007 and it was determined that S. commerson is a carnivorous. More than 99 percent of gut content was different teleost fishes. Gastro somatic index had two peaks in Nov & Jan (before spawning) and with a decreased trend in July, the spawning period. Occurrence of empty stomach was estimated % 65.77. Maximum amount of condition factor was in Dec. Spawning season was started from June. The average of Absolute & relative fecundity (to weight unit) was 1217149±179315 and 178.2±15.58 respectively. Lm50% was estimated 75 cm for females. Sex ratio was 0.97: 1 (male: female). Chi- Square test showed no significant difference (p>0.05). Maximum amount of hepatosomatic index was estimated in March.Metal concentrations were determined using either Flame Atomic Absorption Spectroscopy (for Fe) or Graphite Furnace Atomic Absorption Spectroscopy (for Pb,Cd,Ni and V). The mean concentration (μg/g dry weight)of Pb,Cd,Ni,V and Fe in the liver were 0.0309, 0.0268, 0.0672, 0.0077, 2.5159 in the gonad 0.0440 ,0.0295, 0.1096, 0.0000, 1.4449 and in the muscle 0.0244, 0.0324, 0.0656, 0.0128, 1.6138 respectively. The maximum metal concentrations were below the maximum permissible limits for human consumption recommended by the USEPA, WHO and the UK. The results of Kendall's Tau-b correlation coefficient were as follows: The Liver tissue: There were significant positive linear relationships between accumulation of V, Fe, and Pb with Fork length, Pb and Fe with weight, GSI with Pb, Cd, V and 109 Fe, and a negative linear relationships between HSI with accumulation of V and Fe, Fork length, weight and GSI. The Gonad tissue: There were significant positive linear relationships between GSI with accumulation of Pb, Cd, Fe, Fork length and weight, a negative linear relationship between HSI with Fork length, weight and GSI. The Muscle tissue: There were significant positive linear relationships between accumulations of V, Fe with Fork length and weight factors and as well as GSI with Cd, V, Fe, Pb, Fork length and weight,a negative linear relationship between HSI with Fork length, weight, Cd, Fe and GSI. The results of Mann-Whitney U tests (P≤0.05) show that there were significant differences between summer and autumn from heavy metal contents in the studied tissues point of view. The only exceptions were for Ni in the liver, gonads and muscle and as well as there were significant differences between male and female from heavy metal contents in the studied tissues. The only exceptions were for Pb in muscle, Ni in liver, gonad and muscle, V in muscle, and Cd and Fe in gonads.

Study on some biological and feeding factors of spined loach Cobitis taenia from the Babolrud, Talar and Siahrud Rivers

Spined loach, Cobitis taenia, is a predominant fish in the river systems of the southern Caspian Sea basin. Although there is evidence of the geographical divergence of this taxon, but no information is available on morphological differences within the species populations. This study was designed to evaluate some biological factors including; morphometric and meristic characters, length-weight, age-growth, condition factor, diet, reproduction, variation and differentiation, in the Babolrud, Talar and Siahrud Rivers in south of the Caspian Sea basin. Age, sex ratio, fecundity, ova diameter and gonadosomatic index were estimated. Also, regression analyses was tested the relation between fecundity and fish length, weight, gonad weight, and also age. Totally 858 fish of which 721 were matures, were collected from these rivers by electrofishing. 37 morphometric characters, 9 meristic characters and 78 truss network system characters were estimated. Resulats of DFA analysis based on data of morphometric and meristic showd that these populations are highly (94.5%) varios from each other. In discriminate function analysis, the proportion of individuals correctly classified into their original groups was 61%, 65.4% and 86.5% for upstream and downstream, respectively. Clustering based on Euclidean distances among groups of centroids using an UPGMA and also principal component analysis’ results for morphometric data indicated that these populations from these three rivers were clearly distinct from each other. Regression equations between length and weight in these three populations were significantly different from Folton factor (b=3), that showed the fish has a negative Alometric growth process. Condition factor was estimated between 0.8912 to 1.2736 and 0.8131 to 1.4489 for males and females, respectively. Sex ratio (female: male) in these populations was 1.2816:1. The difference between the number of females and males was significant and females were more than males. The female and male specimens reach maturity by Tl more than 40 and 30 mm and at the age of 2+ and 1+, respectively. The mean of ova diameter was 0.5824±0.2882. The spawning took place from May to late July, at the water temperature from 18.7 to 24.0°C. The GSI values average at the beginning of the reproduction period was about 9%, with ranged from 2 to 26% in ripe mature females. The absolute and relative fecundity were 2109±792 and 579±208 respectively. The absolute fecundity was significantly related to body weight and gonads weight. Based on the pattern of gonado-somatic index, it was concluded that this fish has prolong active reproductive period, which is a type of adaptation by short-lived small fishes to environmental conditions. The macroscopic and histological results showed that the female and male have 5 and 4 stages in their maturation process, respectively. The RLG index was about 0.4732, which showed the fish is a carnivorous species. Significant difference was observed between fishes with different length and diet. The main foods of the fish were Trichoptera, Chironomidae larvae and Ephemeroptera which were their prefered food as well, however it was estimated that the food selection and diet are affected by environmental conditions.

Reproductive biology of Catla catla in the Udawalawe reservoir, Sri Lanka

The reproductive biology of Catla catla (Hamilton-Buchanan) in the Udawalawe reservoir was studied from June 2007 to December 2008. Samples of eggs from Indian major carp C. catla were collected from fish landed in the reservoir and analysed in the laboratory to assess the reproductive characteristics. C. catla. Cirrhinus mrigala, exotic Cichlids and Labeo rohita accounted for 62.2%, 21.0%, 12% and 1.0% respectively of the total landings in the Udawalawe reservoir during the study period. Gonads of C. catla were collected in the field and examined in the laboratory to determine the stage of maturity and fecundity. Data on fish length and gonad weight were collected to estimate the gonado-somatic indices (OSI). Landed catches were also examined in the field to determine the sex ratio of C. catla in the catch and was found to be 1:5.6 male to female. Results of fecundity estimates revealed that C. catla females in the Udawalwe reservoir were fully mature in June and October of the year. From the monthly variation of OSI, two recruitment pulses per year were evident. The length of the body at first maturity in female C. catla in the Udawalwe reservoir was estimated to be 74.2 cm.

Distribution and growth biology of rock oyster Saccostrea cucullata in the Oman Sea

Distribution and growth biology of rock oyster (Saccostrea cucullata) in the northern shores of Oman sea have been struied. During this one-year study, samples have been taken monthly from ten different stations. quantity of vertical distribution of this species was obseredl in the mid - intertidal zone. After determining the spread pattern, the following subjects were studied: - Growth parameters - Distionction of the "cohorts" - Determination of "spawning Season" - Condition of the "Gonado Somatic Index" - Sex ratio - Length of the species during the first year of maturation. - Identification and determination of percentage of "Biofouler Organisms." Results obtained from the above - mentioned studies show that considering a growth factor (k) of 0.52, the value of "Loo " for this species is equal to 114 (mm).Five to six different age groups were observed among the samples taken. In the areas where this study was conducted, this species grows 24 to 30 (mm) in the first year of its life this growth rate is lower in the higher - aged grpups relative to the lewer - aged groups, so that the longest size classes grow between 4 to 6 (mm) per year. • The maxinum Value of the "Condition Index" is in the pozm area and the minimum value of it belongs to Darak and Tang areas. Along with the increase in the growth of gonads the above mentioned condition index increases gradually simultaneous with the onset of spawning. Also, study of the influence of environmental factors on the maturation process suggests that the most important factors affecting maturation and spawing are temperature and salinity. The study of GSI shows that this species has a coordinated bimodal spawing trend, with its spring peak in june and its autumn peak, being still higher than the spring peak, in september. The recruitment curve confirms the above spawning peaks with its peaks occuring after a delay of one month or maximum two months in comparison to the spawning peaks. The results of calcuation of "Sex Ratio" of this species in each area show that sex ratio is 1:1. Among the first size classes that reach maturity, nearly 67% of the samples are male and the remaining 33% are female. with the increase in the shell size, the percentage of males decreases and the percentage of females increases. , The above facts prove the protandrous nature of this species the diagram showing the sizes of the first samples which reach maturity suggests that more than 50% of the samples mature after their length exceeds 36 (mm). The shortest mature sample was found to have a length of 22(mm). After studying "Biofouler Organism" nine different invertebrate groups were indentified. Barnacles and Tunicates have the highest and lowest percentages respectively. According to zonal observations, Barnacles and polychacta do the greatest damage to this species.

Fishery biology of Indian mackerel in Sri Lankan coastal waters

The objective of the project was to determine the reproductive biology of Indian Mackerel (Rastrelliger kanagurta) using Gonado-Somatic Index (GSI); length at first maturity; spawning season; morphological characteristics; and stomach contents.