Morphological and molecular study of new record species of barnacle amphibalanus amphitrite (Darwin, 1854) in BabylonProvince, Iraq.
Amphibalanusamphitrite (previously named Balanusamphitrite,(Clare and Hoeg, 2008) is a major marine biofouling invertebrate distributed in tropical and warm temperate worldwide waters (Clare and Alred 2009), it is belong to balanidae family, and the common names is striped barnacle or purple acorn barnacle, morphological characteristics are a medium-sized, parietal tubes in single row, without transverse septa, exterior of shell with longitudinal striations, shell conical, white and distinctly marked with thick, composed of 6 parietes. Tergum with carinal margin protuberant in upper half to third, spurlength a tenth shorter than width, distance from basiscutal angle less than own width (Gordon, 1970, Biccard, 2012), the classification of this species as follows:
Kingdom: Animalia Phylum: Arthropoda Subphylum: Crustacea Class: Maxillopoda Infraclass: Cirripedia Order: Sessilia Family: Balanidae Genus: Amphibalanus Species: A. amphitrite (Darwin, 1854)
A. amphitrite is a common coastal and estuarine organism present on hard natural surfaces such as bedrock, boulders, mollusc shells and red mangroveroots, It is also available on artificial surfaces such as the hulls of ships, pilings and seawalls, It can be high intensity of growth, with over three hundred individual barnacles being recorded on a single oyster Crassostreavirginica (Boudreauxand Walters, 2005).
A.amphitrite is a common fouling barnacle distributed globally in tropical and subtropical waters (Chenetal., 2014). This species recorded in the south Iraqi saltwater and the Arabian Gulf before many decades (Abdul-Sahib, 2005), and caused of a lot of problems and disadvantages for the cooling system of water intakes of electrical power stations in e Basrah and Nasyria, south of Iraq (Potrous, 2013) and Jaweir& Al-Kenzawi (2009) were recorded this species attached on macrophytes in southern marshes of Iraq as well as Akbar et al.(2012) deal with the study of the effect of some environmental factors on adult density of the barnacles Balanusamphitriteamphitriteat the water intake station Najibiyah Electric Power in Basrah governorate .
No studies recorded the curstean species Amphibalanusamphitrite the aquatic systems in middle of Iraq .While, som studies were used the shells of barnacle as adsorbans materials to remove of some pollution such as industrial dyes and chemical pesticides (Salman et al., 2015 a,b).
This study aimed to identified of barnasle species (A. amphitrite) as a new record in first time on aquatic systems in middle of Iraq by many methods.
MATERIALS AND METHODS
The samples were collected from four stations on the Al- Shomali drainage which connects with Al- Hilla River in west side and Al Masab Alaamdrainagein east side, station 1 selected on the Al-Shomali pumping station, Stationt2 after about 6 km east of station 1(to the Almasab Alaam drainage), station 3on the south of station1 by 1km (before station 1) and station 4on 25 km the northwestern of station 1 by 25 km (like age with Al- Hilla river). Water temperature, pH, dissolved oxygen and salinity were measured also in field accord in to (APHA, 2005).The samples of courtesan species were collected by hand with rock and aquatic plants that attached with from study stations (Fig.2), and transport to laboratory by special plastic bags.
[FIGURE 1 OMITTED]
Taxonomic keys were used to detect morphological characteristics of Amphibalanusamphitrite according to (Gordon, 1970,Henry and McLaughlin, 1975, Kozloff, 1996 and Chanet al.,2009) .The specimen was photos and drawing by digital camera type (Nikon, Japan), Plate3.
[FIGURE 2 OMITTED]
Molecular method was used to confirm the matching of this species through amplification of mitochondrial 12S-rRNA gene as following protocol, Total genomic DNA was extracted using DNA extraction Kit (CAT# A1120) used to identify DNA and we followed the protocol clarified by Promega Corporation, Madison, WI, U.S.A, and after extraction, DNA samples visualized by Electrophoresis then Polymerasechain reaction (PCR) was used to amplify a region of the mitochondrial 12S-rRNA gene from the DNA using forward primer 5'-GACCGTGCTAAGGTAGCATAATC-3' and backward primer 5'-CCGGTCTGAACTCAAATCGTG-3 ', Amplification was done using Master mixture provided by (Bioneer Comp.Korea) which containing (Accu Power[R] PCR Pre Mix, 0.2 ml thin-wall 8-strip tubes with attached cap / 96 tubes, 20 ul reaction): consist of 1-Top DNA polymerase 1 U, 2-Each: dNTP (dATP, dCTP, dGTP, dTTP) 250 [micro]M, 3-TrisHCl (pH 9.0) 10 mM, 4-KCl 30 mM, 5-Mg[Cl.sub.2] 1.5mM. PCR conditions were set as follows: 2 min and 30 sec at 94[degrees]C for initial denaturation, then 30 cycles of 30 s at 95[degrees]C, 30 s at 48[degrees]C, and 1 min at 72[degrees]C, with final extension for 5 min at 72[degrees]C (Wong et al., 2014) and by using Techne Prime G Gradient Thermal Cycler, 96 x 0.2 mL, U.K.
Important characteristics was used to identify this species such as carapace length, height, posterior angle and length-to-height ratio in addition to carapace sculpturing, The barnacle species are limited but it is difficult to identify the species of larvae by their morphological features(Abdul-Sahib, 2005).
A. amphitrite classified according to Gordon (1970), Henry and McLaughlin (1975), Kozloff (1996) and Chan et al. (2009), This species characterized by the wall surface smooth with six plates; Primary parietal tubes with transverse septa; exterior of shell with longitudinal purple striations, transverse teeth on sutural edge with denticles on lower region; basis calcareous and uni-tubiferous; inner surface of scutum with conspicuous adductor ridge. Growth lines in tergum spur exhibiting obvious changes in direction. the A. amphitrite high (14 [+ or -] 2 mm length of rostrum),the base plates diameter was (17 [+ or -] 3 mm), the scutum width (5 [+ or -] 2 mm), the scutum length (6 [+ or -] 2 mm), the tergum width (4 [+ or -] 1 mm) and the tergum length (2 [+ or -] 1 mm) (Plate 1).
This is the first record for A. Amphitrite in Babylon province and meddle of Iraqi, during the sampling the water temperature was (22 [+ or -] 3), salinity was (4 [+ or -] 0.2 0%), pH was (8.2 [+ or -] 0.2),and dissolved oxygen was (9 [+ or -] 1 mg/1).Qiu and Qian (1999) suggest that low temperature and low salinity might be responsible for the decline in reproduction of (Balanusamphitriteamphitrite) A. amphitrite. Akbar et.al.(2012) study the effect of some environmental factors on the density of the adult of the barnacles south of Iraq and they found a positive relationship between adult density and salinity, pH and dissolved oxygen in water, while an inverse relationship between adult density and water temperature.
Many factors may be help to distribution of barnacle in middle region of Iraq such as environmental disturbance of aquatic system and change of many water properties as salinity, temperature, dissolved oxygen, water level ,geological change of sediments and municiapel activities (Qiuand Qian, 1999).The results were showed that the barnaclesdensity increased in the station 2 and 3more than station 4, may be dueto the effect of Al- MasabAlaam drainage near these stations and also, the effect of temperature, salinity, food type and food concentration on larval development (Thiyagarajan et al., 2003) .The distribution of invertebrate fauna living on macrophteroots, epibionts, is partly determined by water flow and distance from the source population (Starczak et al., 2010).
Mitochondrial 12S-rRNA gene was detected in different samples (Figure 2) which have firstly diagnosis by morphological methods according to taxonomic keys, this figure below is illustrated the pcr product of mitochondrial 12S-rRNA gene which amplified by thermal cycler for different samples of A.amphitrite, Amphibalanusamphitrite is a common fouling barnacle dispersed globally in tropical and subtropical waters (Chenet.al., 2014) The 12S-rDNA region has considered to be a successful and valuable tool for barnacle identification, The 12S-rDNA fragments are shorter and relatively easier to amplify by PCR (Wang et al., 2014).
[FIGURE 3 OMITTED]
--The study was recorded the Christian species Amphibalanus Amphitrite Darwin, 1854 in first time as a new record species in aquatic systems in middle of Iraq, Babylon provinous.
--Morphological and Molecular studies were used to identified speciesA. amphitrite.
--The present study was recorded the speciesA.amphitriteas aquatic invasive species in Iraqi aquatic systems
Received 23 July 2015
Accepted 28 August 2015
Available online 25 September 2015
The authors would like to grateful Colleg of Science, University of Babylon for their support to this research
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Adi J.Abd Al-Rezzaq, Ayad M.J. Almamoori, Moayed J. Yass and Jasim M. Salman
Department of Biology, College of Science, University of Babylon, Hilla, Iraq.
Corresponding Author: Adi J.Abd Al-Rezzaq, Department of Biology, College of Science, University of Babylon Hilla,Iraq.
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|Author:||Rezzaq, Adi J.Abd Al-; Almamoori, Ayad M.J.; Yass, Moayed J.; Salman, Jasim M.|
|Publication:||Advances in Natural and Applied Sciences|
|Date:||Aug 1, 2015|
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