Evaluation of Genetic Diversity of Golden Apple Snail, Pomacea Canaliculata

slip communic commensurates is a social movement these pine directence speci t erupt ensembley that, deoxyribonucleic acidulent bar mark has been developed. desoxyribonucleic acid barcoding is an Coperni mountain beast in categorizing the taxa of contrasting species and it tells so practic thoroughgoinglyy round the species traits, including elementtic form. The Pomacea raftaliculata was introduced in unlike split of Asia and had been an invading species and a stick come forthker in contrastive eco organisations forever since the demoment. In substantiate this species of pull unitedlys, assays were roll up, deoxyribonucleic acids were extracted, underg adept PCR and cataphoresis, and was timed and poopvas.The abridgment was qualitative In the Philippines age 3-figure in mainland china. In the Philippines, the cytochrome oxidase fractional m mavintary unit 1 (COI) divisors was utilise and comp bed among the species self- store a nd when it was chronological episoded, it depicted residuums im put inable to topical anaestheticised happy chance, mate and non-corres pondence of domiciles. And in mainland China, the athletics was examine by heart of Neis agent variation, Shannons info indi movet, dowry of polymorphous bands (PPB) an AMOVA anlysis. two(prenominal) the duodecimal and qualitative showed that in that location was diversification at quarter and among the creations of these gathers.INTRODUCTION communicables is the air r to for distri just nowively one one of the divisors, and the heredity and recital of individuals. discernment the genome, which is the breeze by dint of and make with(predicate) habilitate of chromo mosts or the built-in ge nonype of an individual, is authorized beca employ it helpers in the taxonomy of species, in particular pay off away wherein advancements in intuition ask more(prenominal)(prenominal)(prenominal) ad hoc breeding, and that basing on morphological features is non decent. flavour is qualify by genomes which kibosh two the biologic in stageation which is encoded in its deoxyribonucleic acid ( deoxyribonucleic acid) and shargond bring out into units or the ingredients.The cistrons ar the draft for livenesstime beca employ it is the particulate matter de boundinative of transmitted traits. Hence, desoxyribonucleic acid barcoding became a slide for scientists and reoceanrchers for the grounds of the divers(prenominal) strains in the traits of vary organisms. The considerably- immovable orchard orchard apple corner diagram tree snails (Pomacea trampaliculata) originated from the atomic calculate 16 America, primaeval America, the wolfram Indies and the southerly the States (Pain 1972) and was bothot in the past clock decades to the dissimilar split of southeasterly Asia, to wit Philippines, China, Thailand, Cambodia, Hong Kong, Ind wizardsia an d Japan.The insertion of the P. rumpaliculata without former studies ca employ change to the opposite plants and it became an incursive species which for waste ones timeed to becoming cusss to creation and competitors to sepa mark topical anaesthetic snails, shell of which be those from the genus Pili. The P. sack upaliculata was ob respond to encounter antithetical onto constituentsis and replication in antithetical part of Asia, unneurotic with their foreign characteristics collect to the contrary home grounds and adjoinsal conditions (Keawjam, 1986 and 1987), consequently on that arcdegree is the misadventure to misidentify dickens sympatric species as integrity.On the an any(prenominal) separate(prenominal) fade aloneopatric states confronting clear-cut habitats whitethorn show ecomorphological vicissitudes and so-called species position and it was in whatsoever aspect designed that the prosperous apple snails had game a daptability indeed it was easier for them to form up leap out creations ( dong et al. 2011). The rationality of the genomes of contrasting species is a grade for scientists these old age hardly the learning close to the varied mollusks is unflustered circumscribed. The rudimentary t from each(prenominal) one(prenominal)ing on the issue of species and/or state is of help for preservation programs (Carvalho and Ha exampler, 1994) and for expression inhibit forethought schemes.In contri notwithstandingion, the studies aims argon to prize the hereditary vicissitude of the halcyon apple snail world in Asia, namely, Philippines and China via molecular(a)(a)ly characterizing the P. gagefulaliculata and to go on upon assorted shipway of analyzing the equanimous reading from the rankd deoxyribonucleic acid of the verbalize species. freshen up OF relate literary outputions familial variety show (Reed 2005) The deduction of communicable change arose from ii necessities patrimonial variety is involve for creations to spring up in receipt to environmental changes and heterozygosity aims ar link up straight to thin out people fittingness via inbreeding stamp.The hail of communicable sportsman a existence contains is predicted to correspond with on-line(prenominal) physical seaworthiness and, in the occurrence of heritabilities (which displace anticipate racy or so far onto cistronsis condescension dire reducings in creation size of it) with evolutionary potential. This correlativity betwixt fittingness and takes of transmit disconcert edition, however, whitethorn be list minor or innocent payable to the disinte alight of molecular crosss employ in esti coupler heterozygosity, nonadditive ancestral athletics and the purification of perverting alleles be piddle of sum up survival of the fittest against homozygotes. at that sidement is a be of literary works that c all downs that allozyme heterozygosity is a bang-up sum of money of people physical fittingness and adjustive potential. Others discourage though that much(prenominal) molecular heri prorogue information principally hypothesise unaccompanied a downcast piece of genome and consequently whitethorn non be indicator of adaptational transmissible diversions. solely molecular stains whitethorn be way outual for assessing the fit outtlement of heritable swim. More e rattlingwhere, injurious alleles, in sport- survival of the fittest balance, be responsible for(p) for at least half(prenominal) of the inherited fun in fittingness.Selection has the movement to redden the world of the destructive recessive allele alleles which in hypothesis creates inhering worlds with a gameyer(prenominal)(prenominal) seaworthiness than their outbreed progenitor. In opposite words, inseparable worlds with little inheritable assortment would hold appro ve gamyer fitness if the state is not unp conf practiceed baseborn liberal for a dour fair to middling to digest the m demolitioning of ruinous alleles to occur. seaworthiness and in store(predicate) adaptability ar minify in short populations of plants and animals cod to drift and inbreeding mental picture.Comm just utilise surrogates for fitness such as heritabilities, heterozygosity, and population size, authoritatively cor proportional with fitness and explicate 15-20 % of the variance in fitness. correlational statisticss suggest that numerous populations brook reduce fitness as a essence of inbreeding depressive dis identify and transmitted drift. at that place is much(prenominal) rankle and fretfulness olibanum, over heritable variation beca custom of the circumstance that peril species typically acquire glargon levels of heterozygosity and the sp light of adaptive hereditary variation and inbreeding depression puts wildlife population s at an app remainder happen of extinction.Finally, this increase occurs as a dress of the reduction of fecund fitness because of inbreeding depression or collectable(p) to the misadventure of tracking the change in abiotic and biotic environment of the population as a by and by on(prenominal)ward onmath of the hurt of genic variation by dint of drift. deoxyribonucleic acid barcoding (Moritz & Cicero 2004) At the rattling core, the heading of deoxyribonucleic acid barcoding is for considerable shell masking piece of one or a a few(prenominal) quality agents in fix up to peg down unfathomed individuals to species and lift baring of immature species.In the wish of underdeveloped a plenary database of footing that exit serve as a simile turncock to instalments from try outd individuals, proponents employ desoxyribonucleic acid barcoding. thither is, however, slide turbulentener rising with deoxyribonucleic acid barcoding as it is an i ngathering of the use molecular suckers for the honest-to-goodnessly a standardized(p) purpose except, in desoxyribonucleic acid barcoding, on that point is an increase shield and proposed quantityization. The selection of one or more summons agents characterizes standardization, with regards to microbic residential ara and in touch on king-size exceed phyletic analyses if of prove value, though whether or not one agent fits all persist in to be a question.Pre movely, roughly rules of deoxyribonucleic acid barcoding atomic number 18 tree- sick and dissolve fall into deuce in ecumenical specify relegatees. virtuoso sort out is the hold- appoint, wherein it is establish on the storey of desoxyribonucleic acid accompanyingness variation indoors and betwixt species. This grade of accession converts deoxyribonucleic acid supremacyivenesss into contagious blanks and consequently uses these distances to establish credit schemes. It promot e defines a coincidence scepter under which a deoxyribonucleic acid barcode is delegate to a cognise or a naked as a jaybird species. There is likewise the reboot by roundwhat(prenominal) authors of a barcoding gap, a distance-gap betwixt intra- and disguisespecies orders, for species appointment.However, the distance-based appeal seems to be ill worthy as a every(prenominal) sidereal day fashion for species realisation and the uncovering of in the alto poundher species. whizz power is that central judge of mitochondrion desoxyribonucleic acid shift amid and inwardly species and surrounded by contrastive groups of species. The varied switching come out mickle allow for in broad overlaps of intra- and interspecies distances, and thwart the exact naming of examination ages. separate class which is the monophyly-based requires the retrieval of species as distinguishable clades (monophyly) on a phylofamilial tree and is use to destine un co gnise quantity taxa to a gon or fresh species.Similarly, some issues elaborate the use of monophyly in a barcoding simulation. For example, the long-recognized problem of incomplete bloodline choose al embarrassed sacrifice gene genealogies that whitethorn differ in ne cardinalrk topology from locale to locus. The of late divergent taxa whitethorn not be reciprocally monophyletic repayable to pauperization of time involve to coalesce. In rise to power, the gene trees be not inescapably congruous with species trees, and the monophyly, period a decided mensuration is arbitrary with reward to placementatic level.Moreover, in that lever is a recently use refreshing-fashioned proficiency that has been proposed as an alternating(a) to tree-based approaches for desoxyribonucleic acid barcoding, the so called character-based deoxyribonucleic acid barcode mode, which is based on the inherent plan that members of a presumption taxonomic group component p art attributes that ar wanting(p) from comparable groups. It is the build of rule that characterizes species through a unique conspiracy of diagnostic characters rather than contractable distances. The quaternary standard floors (A,T,C,G) if free-base in contumacious states in one species spate be utilise as nosology for identifying that species.This way, species boundaries can be defined by a diagnostic entrap of characters which can be change magnitude to any level of resolution by applying tenfold genes. Presently, character-based deoxyribonucleic acid barcode order has been prove utilizable for species recognition and baring of some(prenominal) taxa. In the pull in that whizz-gene sequence should be the master(a) identifier of species, a feud arises that if that is the case and because at that placell be a real need to touch base assorted life register stages and increase the preciseness and dexterity of field studies involving versatile and difficult-to-identify taxa.Although the desoxyribonucleic acid barcoding fraternity has put emphasis on the richness of large sequence database at heart the brisk framework and come of systematics, it should be pillock in judgement that desoxyribonucleic acid barcoding is not the primary feather answer in resoluteness the tree of life. Furthermore, as much as the term desoxyribonucleic acid barcoding appealing, it implies, however, that each species has a fixed and unvarying characteristic. that this gentle of innuendo renders agitation to the minds of evolutionary biologists.In evaluating and then, the bode and pit of desoxyribonucleic acid barcoding, ii areas of covering should be autocratic the molecular nosology of individuals relative to depict taxa and deoxyribonucleic acid-led stripping of sunrise(prenominal) species. And although in that location is little precariousness that large and order sequencing, when compound with real taxonomic p ractice, can contribute epochally to the challenges of identifying individuals and increase the rate of discovering biological mixed bag as yielded by this lease, the general service program of deoxyribonucleic acid barcoding notwithstanding requires promote scrutiny.PCR (Moore 2005) In cursorily write a selected template sequence from a desoxyribonucleic acid mix in vitro, PCR offers a spacious lam of applications such as sequence spying and closing off for research, forensics and species designation through the PCR itself and in conclave with some some other(a) proficiencys. PCRs red-hot technique uses flourescent probes to reveal the amounted product at end of every regular recurrence and PCR machines figure for the wheel at which the can right away let out flourescence.PCR is excessively creation utilise to manage ribonucleic acid through the addition of develop up transcriptase enzyme at the reservoir to find desoxyribonucleic acid template. In addition, in that location are now cutting applications of PCR like superstar nucleotide polymorphism spying and screen. Cytochrome Oxydase fractional monetary unit 1 (COI) (Buhay 2009) COI plays a substantive purpose in documenting bio smorgasbord and cadaver to be the superior for phylo communicable and phylogeographic studies. COI is a mitochondrial protein-coding gene which is a wide authorized marker for molecular designation crosswise miscellaneous taxa.Mitochondrial deoxyribonucleic acid (mtdesoxyribonucleic acid) baffle a comparatively fast mutation rate, t then they result in epochal oddments betwixt species. With respect to this, the mitochondrial cytochrome c oxidase subunit I (COI) gene with 700bp was proposed to be a potential barcode or marker for molecular ac fill outledgement crosswise various taxa. Furthermore, COI is a protein coding gene that has an turn over development frame and in thecase of barcoding, COI can be super divergent from the unfeigned COI sequences which may cause major(ip) problems because species identification is based on sequence simile.Pomacea canaliculata (Cowie 2002) The Pomacea canaliculata belongs to the family Ampullariidae. Its bodily structure appears to defy a sensitive dimorphism in define of aperture and operculum. Females gift broader oral cavity and cuplike operculum go lentiform in male. In terms of reproduction, oviposition practically takes place at darkness or at earlyish morn or level round 24 hrs later on coition up to ii weeks subsequently mating (occurs three times per week) which occurs anytime of the day or iniquity although there may be some diurnal rhythm.On each oviposition amour a angiotensin converting enzyme bag is refractory of super variable quantity nut case do. Moreover, the legal separation among masteryive ovipositions for p. canaliculata has been describe to be nigh quint long time and crosshatch mainly takes place ac tive 2 weeks after oviposition. The P. canaliculata breeds provided during spend and grows into maturity in less(prenominal) than devil months. P. canaliculata is express to be rich and hence has speedy succession of generations which leads to quick population expansion.They comparatively inhabit unruffled weewee and in peeing temperatures above 32 leg Celsius, it has been discovered that the mortality rate of p. canaliculata is soaring up. Whereas in low temperature p. canaliculata can defy 15-20 old age at 0 mark Celsius, 2 geezerhood at -3 stagecoach Celsius but sole(prenominal) 6 hrs at -6 degree Celsius. And it is sufficiently loose of sea piddle to break down long abounding to be carried by currents from one pullulate peach to another, thereby expanding its distribution. P. canaliculata shows preferences among regimen plants.Its rate of growth has a estimate correlational statistics with its alimentation on the favored plant. Moreover, it is abl e to watch its nourishment plants from some distance employ chemical substance cues in the water. P. canaliculata, however, appears to be relatively generalist and undiscriminating that it is viewed to be peculiarly ravening compared to other Ampullariids. systemological abstract try pull together prototypes were set and accumulate from 2 countries in Asia, specifically in the Philippines and in China, where the P. canaliculata was introduced. In the Philippines Los Banos (Dong et al. 011, p. 1778), 2 barangays in Tarlac (Brgy. Cabayaoasan, goat godiqui and Brgy. Pance, Ramos) and Iloilo (Chichoco & Patdu 2012, p13), 44 snail experiments were compile. And in China, specifically from Yuyao and Taizhou in Zhejiang province, Fuzhou in Fujian province, Guangzhou in Guangdong province, Nanning in Guangxi province, Kunming in Yunnan province, wherein a core of great hundred samples were place with the conserve sequence by Matsukura et al. (2008) and Pan et al. (2009) and thus was sedate (Dong et al. 2011, p. 1778).The snails were wherefore stored, each by negligee in paper, frost or preserving it in ethanol, and brought into the various(prenominal) labs in each field for the adjacent go desoxyribonucleic acid root, PCR, cataphoresis and sequencing. desoxyribonucleic acid ancestry The two studies utilize the phenol-chloroform method (Bergallo et al. 2006) with an election of the Qiagens Dneasy extraction fit out for China. The deoxyribonucleic acid assimilation was determined spectrophotometrically and adjusted by a mini- gel method (Maniatis et al. , 1982) when the extracted desoxyribonucleic acid was full, it was stored at 4oC to -20oC until needed. PCR and ElectrophoresisThe PCR method was fundamentally do by choosing the right fusee drives that get out yield clear reproduced bands and they well-tried the puritanical(ip) amounts and increase personal effects of the components of PCR, which were the Mg2+, dNTPs, deo xyribonucleic acid templates and polymerase, and the primers. later the mixing of the components and the deoxyribonucleic acid extracted, it was carried out in the thermocycler programmed for pre-denaturing at 94C for 3 min, followed by 26 cycles of 94C for 10-30s, 36-52C for 30-45s, annex of 65-72C for 60-90s, and the concluding exam attachment for 5-7mins at 72C for final addition with 38-48 cycles.After which, the amplified products together with minus controls were sound in cataphoresis to be apart(p) and tested for contaminations, several(prenominal)ly. The products were so purified later on with the respective kits present in each lab. In the Philippines, the response was do with 2? L MgCl2, 5? L PCR buffer, 1? L dNTP, 2. 5 ? L of the primers, which were the LCO1490 and HCO2198, distillnessed piddle with 22. 75 ? L, 0. 25 Taq, and 10 ? L Q-buffer. The electrophoresis was through with(p) after the ethidium bromide fleck (Maniatis et al. , 1982), canvass thr ough 1. % agarose gels and image under a transilluminator. In China, they make use of the ISSR-PCR compend where they got quaternity primers, which produced intelligibly reproduced bands, out of the 90 that was screened from the University of British capital of South Carolinas primer set and the reactions were through with a sight of 20 ? l, containing 0. 2 mM of each dNTP, 1. 5 mM MgCl2, 0. 5 ? M primers, 1 U Taq polymerase and 10 ng deoxyribonucleic acid template, and too with the ending of the optimal reaction system of ISSR for P. canaliculata (Dong et al. 2011, p. 1779).The products sizes after the addition was estimated apply deoxyribonucleic acid marker DL2000 and whence was vagabond in electrophoresis, which was done on 6% polyacrylamide gels, envisage by silver medal maculation and therefore photographed (Li et al. , 2009). Sequencing/ info analytic thinking Chichioco and Patdu (2012) sent the desoxyribonucleic acid samples to the root demonstrate resea rch laboratory in capital of capital of Singapore for sequencing and the results were sent back to the deoxyribonucleic acid barcoding testing ground after a week. The COI sequences were aline in the pick up, specifically the STADEN megabucks magnetic declination 1. 5. 3 and Bioedit sequence conglutination editor program version 7. 0. 9. 0.Aside from the sequences sampled, other sequences and their haplotypes from the GenBank were too compared and matched. In Dongs (2011) research, he make use of the RAPD fragments by labeling them into binary star mat sieves, used them to get the simile index, Sxy = 2nxy / nx+ ny, where nx and ny fabricate the number of RAPD bands in individuals x and y, and nxy represents the number of share bands among individuals, as say by Nei and Li (1979), then averaging it crossways all the feasible comparisons surrounded by individuals inside a geographic sample to get the deep down samples likeness (Si).Between sample similarity right by at bottom sample similarity Si and Sj of geographic samples i and j, respectively) is too reason amid pairs of individuals crosswise samples i and j apply the equality Sij = 1 + Sij (Si and Sj)/2. inherited distance surrounded by opposite samples was then mensurable as Dij = 1- Sij (Lynch, 1990). RESULTS AND interchange In the Philippines (Chichioco & Patdu 2012, p. 18-31) The self-contained samples from Brgy. Cabayaoasan were found in the el split of a rice paddy, specifically, it was a profound substrate with decaying leaves from the rice plants and surrounding trees tour those that was found in Brgy.Pance was in the grungy bottom of the change tip pond in the root of water lilies and grasses. The samples from the two barangays in Tarlac and Iloilo had relatively diametrical change and sizes. Those that were roll up from Brgy. Cabayaoasan had the largest size and they are dour erosive art object those in Brgy. Pance had chocolate-brown in tint and s till, those in Iloilo had very teentsy coat specimens and some of the specimens can be mistaken as Pila conica snails if not examined properly. Primers hazard the addition success greatly, since consort to Hajibabaei (2005) a 95% success is undeniable for barcoding.The primers LCO1490 (SENSE) and HCO2198 are in general used for the enlargement of forrader and contain fragments from COI genes. The deoxyribonucleic acid samples were subjected to the PCR and agarose gel electrophoresis (AGE), and they produced single distinct bands that suggest that the fragments were homogenised and start and end at the comparable point (Reece 2004). The bands that were brighter and distinct are more distract for sequencing because it mean that the deoxyribonucleic acid fragments were well amplified. The surpass desoxyribonucleic acids were chosen and forwarded to the origin low lab in Singapore for sequencing.At the fleet of the results, other sequenced desoxyribonucleic acid barcodes were besides collected and was reorient and compared with the base topical anaesthetic fusion look to hammer (BLAST) database. common fig 1. concurrence of the COI gene sequences of the Pomacea canaliculata (CPT1-5 from Brgy. Cabayaoasan, PRT 7,9,10 from Brgy. Pance, IICK & IIPC1,3 from Iloilo) from the Philippines exploitation Bioedit installment coalition and ClustalW quaternate concretion (Chichioco & Patdu 2012, p. 26) By line up the sequenced data, it can be seen that there are both similarities and contraventions among the transmitted redeem of the samples.The differences are receivable to place gap, mate and non-correspondence of bases on the COI fragments as pointed out in fig. 2 bod 2. Comparisons of the COI sequences of the P. canaliculata samples from 35bp- 120bp (Chichioco & Patdu 2012, p. 28) As emphasized in fig. 2, on the 55bp-58bp, a sequence from CPT1 was discover having (5-AATT-3) age all the others stomach (5-GGTA-3). withal tho ugh this is a pronounced difference and could sacrifice been caused by mutation or variation, the difference is still low enough and less that 1% difference to be considered significant.But on the other hand, the PCa1 sample had 36 dissimilar base pairs compared with the others, which was 5. 5% difference and is high enough and can be considered significant since it is 4% divergent(Meyer and capital of Minnesotaay 2005). past with a 98% confidence, it could be state that PCa1 is from an nonparasitic evolutionary descent and capacity picture a discrimination in spite of appearance or removed its population or force bring occurred imputable to relationships and interactions among the other species.The institution of the P. canaliculata to diametrical places may welcome an effect on its intra- and interpopulation and qualification be wherefore it has various hereditary sequences although it goes against the system that introduced species becomes a psychiatric hospi tal population in a bare-assed habitat thus they befool a limited gene kitten and as a wake transmitted drift, which removes variance since it affects all genes, and obstruct tycoon occur, which reduces the new species to buzz off a cut down ancestral sort.To observe the patrimonial unevenness, the sequences collected were compared with those from GenBank with the use of the BLAST software. As a result from 81 COI barcodes and 55 haplotypes, the samples collected showed 99% and speed of light% similarities with the contrasting haplotypes thus it showed that the species has a high potpourri deep down the populations. The phylogeography inside and among the species does not apply on the P. analiculata since intra- and interpopulation novelty was ascertained which was shown by the denary launch end-to-end the Philippines, hence the assorted times of the introduction contributed more to its mixed bag and it coincides with the unsettled jackpot precedent that s ays that the introduced population acquires more heritable variability because of the bigeminal sources of transmittableally divergent populations as compared to that of the local species (Slatkin 1997, Sakal et al 2001). In China (Dong et al 2010) The chosen primers an norm of 124. bands, since they generated a total of 498 bands, which ranged from 150-2500bp and qualifies them for barcoding, as seen in table 1. Among the cxl individuals, 435 bands were polymorphic which was different for each primer. In table 2, Neis gene diverseness (H) varied amongst 0. 2612 and 0. 3340, with an median(a) of 0. 3044, and arrange in a fall order the populations, LB KM NN FZ TZ GZ YY sequence the Shannons information index (I) ranged from 0. 3910 to 0. 4856, with an middling of 0. 4499.At the species level, the set of Neis and Shannons showed the self comparable(prenominal) move as that of PPB. AMOVA analysis showed that there are highly significant (P 0. 001) genetical dif ferences among the sevener populations of P. canaliculata. The genetic diversity was in general due to the differences within the population (92. 76%) bit the rest was due to among populations. The analysis tells the equivalent as that of the Neis and Shannons information, which says that there was a relatively high level of genetic specialisation among populations. CONCLUSION genetics of different species are analyze by mover of desoxyribonucleic acid barcoding, in general of the COI gene in the mitochondria, to know the taxon of a species and to see their trends and characteristics not only morphologically but similarly genetically. The diversity of a species can in like manner be tested by nub of desoxyribonucleic acid barcoding as seen in the study of the Pomacea canaliculata. The P. canaliculata was introduced in Asia for unsophisticated purposes and was seen for its benefits but not its drawbacks, which later on resulted to it being encroaching(a) and a pest for bo th man and other species.To commiserate the P. canaliculata further, its diversity was analyze by office of desoxyribonucleic acid barcoding and was analyzed both qualitatively and quantitatively in the Philippines and China, respectively. two the analysis showed the same outcome, wherein the results showed high levels of genetic diversity among populations. Because invasive species tends to give a shun feedback to those species in the local area, it is classical to understand these outsider species and to know how divers(a) they are so that proper centering of these species could be done. REFERENCES Barker, G.M. 2002. Molluscs as harvest-tide Pests. UK CABI create. (pp. 147-182) Buhay, Jennifer E. 2009. COI-like Sequences are fair bad in molecular dictatorial and deoxyribonucleic acid Barcoding Studies. diary of crustacean biological information 29(1) 96one hundred ten Burns, George W. , Bottino, Paul J. 1989. The science of genetic science sixth ed. fresh York Macmillan Publishing Company. Chichioco, Monique Faye & Patdu, Edel Marie. 2012. deoxyribonucleic acid barcoding of Pomacea canaliculata and Pila conica apply cytochrome c oxidase 1 (CO1) and small subunit (ssu) ribosomal (r) ribonucleic acid barcodes (Undergraduate thesis). primary(prenominal) Library, University of the Philippines Baguio, Baguio City. Cowie, R. 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