Fish prey of southern elephant seals, <Emphasis Type="Italic">Mirounga leonina</Emphasis>, at King George Island

Between November and February in the years 1993/1994 to 1999/2000, 153 southern elephant seals, Mirounga leonina, were stomach lavaged at King George Island. Fish occurred in 16 of 108 stomachs containing food items. Myctophids were the dominant fish prey, contributing 76.6% of the 145 sagittal otoliths found. The most important prey species was Gymnoscopelus nicholsi, which constituted 69% of the otoliths and occurred in 75% of stomach samples. Next in importance was the nototheniid Pleuragramma antarcticum, which occurred in 31.3% of samples and represented 11.7% in numbers. We suggest that while myctophids may be the dominant fish prey of elephant seals in areas close to King George Island, they are probably replaced by P. antarcticum as seals migrate towards higher latitudes where this fish is extremely abundant.     

Seasonal use of oceanographic and fisheries management zones by juvenile southern elephant seals (<Emphasis Type="Italic">Mirounga leonina</Emphasis>) from Macquarie Island

The foraging distribution of marine predator populations is important for effective modelling and management of pelagic marine systems. We tracked 31 juvenile southern elephant seals from Macquarie Island (158°57E, 54°30S) over their annual post-moult and mid-year trips to sea. We calculated the amount of time spent in regional fisheries management areas and within bounded oceanographic regions. During the austral summer, juvenile seals spent over 90% of their time south of the Antarctic Polar Front and ~80% within fisheries management regions [Commission for the Conservation of Antarctic Marine Living Resources (CCAMLR) and exclusive economic zones]. In winter, seals spent ~75% of their time in the region bounded by the Antarctic Polar Front and the southern boundary of the Antarctic Circumpolar Current, and ~60% within fisheries management regions. The time spent per region differed significantly between summer and winter. Our results demonstrate that juvenile southern elephant seals from Macquarie Island spent more time south of the Antarctic Polar Front and within fisheries management areas than previously suspected.     

Mercury concentrations in the Patagonian toothfish, <Emphasis Type="Italic">Dissostichus eleginoides</Emphasis> Smitt 1898, among three distinct stocks

Patagonian toothfish, Dissostichus eleginoides, collections (n = 186) from three distinct stocks (two subantarctic stocks and the stock within the Chilean ZEE) were analyzed for Hg concentration by DMA and comparisons were made by gender, total length (TL), wet weight (WW), and population. There was no difference between TL–WW relationships or Hg concentration by gender within any population across the range examined. The Chilean and Prince Edward Island fish had higher TL-adjusted total Hg concentration than those from the South Georgia and may suggest a possible human health risk. Explanation of these population differences in total Hg may be the actual sample areas and associated hydrogeographic and oceanographic conditions. For example, sampling sites for the Chilean fish lie well outside the Antarctic Convergence whereas the Prince Edward sites straddle the Convergence. The samples obtained from South Georgia lie well within the Antarctic convergence, suggesting that the Polar Front may provide some type of hydrographic barrier as has been shown for this species in terms of larval dispersal and feeding.     

Resource partitioning through oceanic segregation of foraging juvenile southern elephant seals (<Emphasis Type="Italic">Mirounga leonina</Emphasis>)

In highly dynamic and unpredictable environments such as the Southern Ocean, species that have evolved behaviors that reduce the effects of intra-specific competition may have a selective advantage. This is particularly true when juveniles face disadvantages when foraging due to morphological or physiological limitation, which is the case for many marine mammals. We tracked the at-sea movements of 48 juvenile southern elephant seals (Mirounga leonina) between the ages of 1 and 4 years from the population at Macquarie Island using locations derived from recorded light levels. There were significant differences in the total amount of the Southern Ocean covered by the different age-groups. The younger seals used a smaller area than the older seals. On average, the younger individuals also made more trips to sea than the older seals and did not travel as far on each trip. Females spent more time at sea than males and there were no significant differences between the total areas used by male and females. In summary, younger seals remained closer to the island at all times, and they spent more time in more northerly regions that older seals. These differences in behavior created temporal and spatial segregation between juveniles of different ages. Therefore, we suggest that these temporal and spatial separations help to avoid intra-specific competition for resources on land, space on beaches, and at-sea foraging areas. Such modifications of haul-out timing and behavior enable them to exploit a patchy and unpredictable environment.     

Mercury in the marine food chain in the Southern Ocean at Macquarie Island: an analysis of a top predator,Patagonian toothfish (<Emphasis Type="Italic">Dissostichus eleginoides</Emphasis>) and a mid-trophic species,the warty squid (<Emphasis Type="Italic">Moroteuthis ingens</Emphasis>)

The characteristics and habitat of the Patagonian toothfish (Dissostichus eleginoides) are typical of fish that accumulate high concentrations of mercury. In this study, mercury determinations were made on samples of muscle tissue from Macquarie Island toothfish and the Southern Ocean deepwater warty squid (Moroteuthis ingens). The analysis of mercury in the biological tissues was made by cold vapour-atomic absorption spectrometry following acid digestion. Performance of the analytical procedure was assessed by analysis of certified reference material (DORM-2, dogfish muscle). Mercury concentrations of 16 Macquarie Island toothfish ranged from 0.12 mg kg^–1 (550 g, 381 mm TL) to 0.59 mg kg^–1 (6,100 g, 823 mm TL), with a mean concentration of 0.33±0.12 mg kg^–1. A significant correlation was found between mercury and either toothfish weight or total length. The fish analysed were juveniles, which suggests that larger individuals would have higher mercury concentrations well exceeding food standard code limits for mercury in fish (typically 0.5 mg kg^–1). Warty squid, also from around Macquarie Island, had a low mean mercury concentration of 0.086 mg kg^–1 in mantle tissue; no significant correlation existed between mercury concentration and either squid mantle length or total weight. It is postulated that the squid have a mechanism, possibly involving the digestive gland, that prevents bioaccumulation of mercury in the mantle, and presumably other body tissues.     

The population trend of southern elephant seals (<Emphasis Type="Italic">Mirounga leonina</Emphasis> L.) at Macquarie Island (1952–2004)

Total numbers of adult female southern elephant seals (cows) breeding at Macquarie Island were determined for 19 of the 52 year period between 1952 and 2004. Totals for 1952–1987 (exc. 1959 and 1985) were estimated from the relationship between censuses of the isthmus study area and concurrent censuses for the whole island. Totals for 1987–2004 were obtained by direct census of the entire island in mid-October. Cow numbers decreased from a maximum of about 40,000 in the 1950s to a minimum of 18,300 in 2000, but then increased slightly to 19,200 in 2004. Nonlinear and post-hoc linear analysis of the count data identified 1999 as the year when the exponential rate of change (r) slowed from −1.4% per annum to near zero. The rate of change was not uniform for each census sub-area counted (1987–2004), suggesting that certain terrestrially based density-dependent mechanisms were influencing the annual distribution of cows.     

Disentangling reproductive biology of the Patagonian toothfish <Emphasis Type="Italic">Dissostichus eleginoides</Emphasis>: skipped vs obligatory annual spawning,foraging migration vs residential life style

Patagonian toothfish Dissostichus eleginoides Smitt 1898, is an important commercially targeted Notothenioid species in South-Atlantic waters. In this study we aimed to clarify several aspects of reproductive biology of Patagonian toothfish in the Falkland Islands waters. Histological examination of female gonads indicates that with the beginning of maturation females maintain at least two populations of oocytes, suggesting that toothfish requires up to two years for oocytes development. Females become mature at an average size of 79.1 cm indicating a decrease of first maturity size if toothfish females in the Falkland Island waters. The majority of females spawn at the size from 101 to 130 cm total length. Distribution of reproductive phases shows an increase of females at developing stage in December and March prior to the spawning peaks in May and August respectively. However, the majority of the toothfish population consist of non-spawning individuals remaining in regressing phase (55.8 to 85.6%) including the spawning period. The skip-spawning for toothfish has been defined as reabsorbing non-reproductive and resting types. The abbreviation of oocytes development in the gonads was observed from 1 to 22.1% of females which omitted the spawning season. Most likely females which remain in the spawning area have the opportunity to spawn more often, whereas females which undergo foraging migration toward Northern parts of the Falklands waters return to the spawning ground less often. Females remain Northern area longer to accumulate necessary amount of energy. This hypnotise is supported by the presence of females in immature, developing and regressing phase throughout surrounding Falkland Islands waters. Presence of post-spawning females in regressing stage throughout the Falklands waters suggest that toothfish may undertake irregular spawning/foraging migration when favourable for spawning condition occur.     

Immobilization of adult male southern elephant seals (<Emphasis Type="Italic">Mirounga leonina</Emphasis>) during the breeding and molting periods using a tiletamine/zolazepam mixture and ketamine

Seventy-seven immobilizations were carried out on adult male southern elephant seals at Stranger Point, Isla 25 de Mayo (King George Island) using a combination of Zoletil^® (tiletamine and zolazepam) and ketamine in order to obtain biological samples. During 2006/2007, 22 males were immobilized at the beginning of their breeding period (EB), 19 of which were recaptured at the end of breeding (LB). Four were given only once at an unknown stage of breeding (USB) and 18 males were immobilized at the beginning of molting (BM). During 2007/2008, 14 adult males were immobilized at an USB. Zoletil^® was administered using an automatic discharge device, whereas ketamine was injected directly with a syringe, and was used only when the initial sedation was not enough to carry out the programmed sampling. The initial mean dose of Zoletil^® was 1,387 ± 304 mg, which represented 0.60 ± 0.14 mg/kg, range 0.36–1.05, n = 77. In 47 procedures, an average dose of 1.04 ± 0.66 mg/kg of ketamine was added. Mean immobilization time was 34 ± 14 min. In 25 out of the 77 procedures, males showed apnea, which lasted 8 ± 4 min (range 2–15 min). The necessary doses of Zoletil^® and ketamine to attain immobilization differed between stages. For animals taken twice, doses (mg/kg) of Zoletil^® and ketamine were significantly higher at the beginning than at the end of breeding. During molting, the doses of Zoletil^® given were significantly lower than those used during breeding, although the proportion of animals that required ketamine during molting was significantly higher than during breeding. Zoletil^® proved to be a safe immobilizing agent for field work on adult males of this species, given the wide range of doses used without any serious consequences. Furthermore, the addition of ketamine was useful when the initial sedation was not satisfactory or for prolonging the immobilization period in a practical and reliable way.     

<Emphasis Type="Italic">STAYGREEN</Emphasis> (<Emphasis Type="Italic">CsSGR</Emphasis>) is a candidate for the anthracnose (<Emphasis Type="Italic">Colletotrichum orbiculare</Emphasis>) resistance locus <Emphasis Type="Italic">cla</Emphasis> in Gy14 cucumber

Key message

Map-based cloning identified a candidate gene for resistance to the anthracnose fungal pathogen Colletotrichum orbiculare in cucumber, which reveals a novel function for the highly conserved STAYGREEN family genes for host disease resistance in plants.

Abstract

Colletotrichum orbiculare is a hemibiotrophic fungal pathogen that causes anthracnose disease in cucumber and other cucurbit crops. No host resistance genes against the anthracnose pathogens have been cloned in crop plants. Here, we reported fine mapping and cloning of a resistance gene to the race 1 anthracnose pathogen in cucumber inbred lines Gy14 and WI 2757. Phenotypic and QTL analysis in multiple populations revealed that a single recessive gene, cla, was underlying anthracnose resistance in both lines, but WI2757 carried an additional minor-effect QTL. Fine mapping using 150 Gy14?×?9930 recombinant inbred lines and 1043 F₂ individuals delimited the cla locus into a 32 kb region in cucumber Chromosome 5 with three predicted genes. Multiple lines of evidence suggested that the cucumber STAYGREEN (CsSGR) gene is a candidate for the anthracnose resistance locus. A single nucleotide mutation in the third exon of CsSGR resulted in the substitution of Glutamine in 9930 to Arginine in Gy14 in CsSGR protein which seems responsible for the differential anthracnose inoculation responses between Gy14 and 9930. Quantitative real-time PCR analysis indicated that CsSGR was significantly upregulated upon anthracnose pathogen inoculation in the susceptible 9930, while its expression was much lower in the resistant Gy14. Investigation of allelic diversities in natural cucumber populations revealed that the resistance allele in almost all improved cultivars or breeding lines of the U.S. origin was derived from PI 197087. This work reveals an unknown function for the highly conserved STAYGREEN (SGR) family genes for host disease resistance in plants.

     

Multiple paternity in <Emphasis Type="Italic">Rhipicephalus</Emphasis> (<Emphasis Type="Italic">Boophilus</Emphasis>) <Emphasis Type="Italic">microplus</Emphasis> confirmed by microsatellite analysis

The aim of this study was to determine if individual ticks among the progeny of a single female Rhipicephalus (Boophilus) microplus tick removed from cattle under natural conditions are the result of mating with one or several males. To this end, simulations were run using an existing dataset of genotypes from 8 microsatellite loci to predict the number of samples required and the best locus. Subsequently, 14–22 progeny from each of 15 engorged female ticks removed from three cows, and the engorged females themselves, were genotyped for the BmM1 locus and the minimum number of potential male parents was determined for each progeny group. Of the 15 progeny groups, 10 must have been sired by more than one male, as indicated by the presence of five unique alleles among the progeny or three unique alleles that could not have been contributed by the female. This finding demonstrates multiple paternity in R. microplus.     

Evaluation of the toxicity of <Emphasis Type="Italic">Arthrospira</Emphasis> (<Emphasis Type="Italic">Spirulina</Emphasis>) <Emphasis Type="Italic">platensis</Emphasis> extract

In this study, the methanol extract of Arthrospira (Spirulina) platensis was examined for acute and subchronic toxicities. The extract did not produce any sign of toxicity within 7 days after feeding it at a single high dose of 6 g kg⁻¹ body weight to female and male Swiss mice. For the subchronic toxicity test, the extract at doses of 6, 12, and 24 mg kg⁻¹ body weight was orally administered to six male and six female Wistar rats daily for 12 weeks. Throughout the study period, we did not observe any abnormalities on behavior, food and water intakes, and health status among the treated animals. The hematology and clinical chemistry parameters of treated groups did not significantly differ from those of the controls in both sexes. Postmortem examination of the test groups also showed no abnormalities in both gross and histological findings. These results thus suggest that the methanol extract of A. platensis did not cause acute or subchronic toxicity in our experimental animals.     

First record of the tick <Emphasis Type="Italic">Ixodes</Emphasis> (<Emphasis Type="Italic">Pholeoixodes</Emphasis>) <Emphasis Type="Italic">kaiseri</Emphasis> in Turkey

Nymphs and larvae belonging to Ixodes spp. were collected from a red fox in Turkey. The ticks were identified morphologically and molecularly (16S rDNA PCR and phylogenetic analysis) as I. kaiseri. Sequence and phylogenetic analyses show that our I. kaiseri isolate is very similar to I. kaiseri isolates collected from Germany, Serbia, Romania, and Hungary. Therefore, the existence of I. kaiseri has been demonstrated for the first time in Turkey. More studies relating to the regional distribution and vectorial competence of I. kaiseri are needed.     

Actinomycetemcomitin: a new bacteriocin produced by <Emphasis Type="Italic">Aggregatibacter</Emphasis> (<Emphasis Type="Italic">Actinobacillus</Emphasis>) <Emphasis Type="Italic">actinomycetemcomitans</Emphasis>

Aggregatibacter (Actinobacillus) actinomycetemcomitans P_7–20 strain isolated from a periodontally diseased patient has produced a bacteriocin (named as actinomycetemcomitin) that is active against Peptostreptococcus anaerobius ATCC 27337. Actinomycetemcomitin was produced during exponential and stationary growth phases, and its amount decreased until it disappeared during the decline growth phase. It was purified by ammonium sulphate precipitation (30–60% saturation), and further by FPLC (mono-Q ionic exchange and Phenyl Superose hydrophobic interaction) and HPLC (C-18 reversed-phase). This bacteriocin loses its activity after incubation at a pH below 7.0 or above 8.0, following heating for 30 min at 45°C, and after treatment with proteolytic enzymes such as trypsin, α-chymotrypsin, and papain. Actinomycetemcomitin has a molecular mass of 20.3 KDa and it represents a new bacteriocin from A. actinomycetemcomitans.     

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.     

<Emphasis Type="Italic">Sex combs reduced</Emphasis> (<Emphasis Type="Italic">Scr</Emphasis>) regulatory region of <Emphasis Type="Italic">Drosophila</Emphasis> revisited

The Hox gene Sex combs reduced (Scr) is responsible for the differentiation of the labial and prothoracic segments in Drosophila. Scr is expressed in several specific tissues throughout embryonic development, following a complex path that must be coordinated by an equally complex regulatory region. Although some cis-regulatory modules (CRMs) have been identified in the Scr regulatory region (~75 kb), there has been no detailed and systematic study of the distinct regulatory elements present within this region. In this study, the Scr regulatory region was revisited with the aim of filling this gap. We focused on the identification of Initiator elements (IEs) that bind segmentation factors, Polycomb response elements (PREs) that are recognized by the Polycomb and Trithorax complexes, as well as insulators and tethering elements. To this end, we summarized all currently available information, mainly obtained from high throughput ChIP data projects. In addition, a bioinformatic analysis based on the evolutionary conservation of regulatory sequences using the software MOTEVO was performed to identify IE and PRE candidates in the Scr region. The results obtained by this combined strategy are largely consistent with the CRMs previously identified in the Scr region and help to: (i) delimit them more accurately, (ii) subdivide two of them into different independent elements, (iii) identify a new CRM, (iv) identify the composition of their binding sites and (v) better define some of their characteristics. These positive results indicate that an approach that integrates functional and bioinformatic data might be useful to characterize other regulatory regions.     

Bio-resolution of glycidyl (<Emphasis Type="Italic">o</Emphasis>, <Emphasis Type="Italic">m</Emphasis>, <Emphasis Type="Italic">p</Emphasis>)-methylphenyl ethers by <Emphasis Type="Italic">Bacillus megaterium</Emphasis>

A newly isolated Bacillus megaterium with epoxide hydrolase activity resolved racemic glycidyl (o, m, p)-methylphenyl ethers to give enantiopure epoxides in 84–99% enantiomeric excess and with 21–73 enantiomeric ratios. The (S)-enantiomer was obtained from rac-glycidyl (o or m)-methylphenyl ether while the (R)-epoxides was obtained from glycidyl p-methylphenyl ether. The observations are explained at the level by enzyme-substrate docking studies.     

<Emphasis Type="Italic">In silico</Emphasis> and <Emphasis Type="Italic">in situ</Emphasis> characterization of the zebrafish (<Emphasis Type="Italic">Danio rerio</Emphasis>) <Emphasis Type="Italic">gnrh3</Emphasis> (sGnRH) gene

Background

Gonadotropin releasing hormone (GnRH) is responsible for stimulation of gonadotropic hormone (GtH) in the hypothalamus-pituitary-gonadal axis (HPG). The regulatory mechanisms responsible for brain specificity make the promoter attractive for in silico analysis and reporter gene studies in zebrafish (Danio rerio).

Results

We have characterized a zebrafish [Trp⁷, Leu⁸] or salmon (s) GnRH variant, gnrh 3. The gene includes a 1.6 Kb upstream regulatory region and displays the conserved structure of 4 exons and 3 introns, as seen in other species. An in silico defined enhancer at -976 in the zebrafish promoter, containing adjacent binding sites for Oct-1, CREB and Sp1, was predicted in 2 mammalian and 5 teleost GnRH promoters. Reporter gene studies confirmed the importance of this enhancer for cell specific expression in zebrafish. Interestingly the promoter of human GnRH-I, known as mammalian GnRH (mGnRH), was shown capable of driving cell specific reporter gene expression in transgenic zebrafish.

Conclusions

The characterized zebrafish Gnrh3 decapeptide exhibits complete homology to the Atlantic salmon (Salmo salar) GnRH-III variant. In silico analysis of mammalian and teleost GnRH promoters revealed a conserved enhancer possessing binding sites for Oct-1, CREB and Sp1. Transgenic and transient reporter gene expression in zebrafish larvae, confirmed the importance of the in silico defined zebrafish enhancer at -976. The capability of the human GnRH-I promoter of directing cell specific reporter gene expression in zebrafish supports orthology between GnRH-I and GnRH-III.