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package prok;
import java.io.PrintStream;
import java.util.ArrayList;
import java.util.Arrays;
import java.util.concurrent.locks.ReadWriteLock;
import java.util.concurrent.locks.ReentrantReadWriteLock;
import aligner.SingleStateAlignerFlat2;
import fileIO.ByteFile;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import shared.Parse;
import shared.Parser;
import shared.PreParser;
import shared.Shared;
import shared.Timer;
import shared.Tools;
import stream.ConcurrentReadInputStream;
import stream.ConcurrentReadOutputStream;
import stream.FastaReadInputStream;
import stream.Read;
import structures.ListNum;
import template.Accumulator;
import template.ThreadWaiter;
import tracker.ReadStats;
/**
* Splits a mix of ribosomal sequences (such as Silva) into different files per type (16S, 18S, etc).
*
* @author Brian Bushnell
* @date November 19, 2015
*
*/
public class SplitRibo implements Accumulator<SplitRibo.ProcessThread> {
/*--------------------------------------------------------------*/
/*---------------- Initialization ----------------*/
/*--------------------------------------------------------------*/
/**
* Code entrance from the command line.
* @param args Command line arguments
*/
public static void main(String[] args){
//Start a timer immediately upon code entrance.
Timer t=new Timer();
//Create an instance of this class
SplitRibo x=new SplitRibo(args);
//Run the object
x.process(t);
//Close the print stream if it was redirected
Shared.closeStream(x.outstream);
}
/**
* Constructor.
* @param args Command line arguments
*/
public SplitRibo(String[] args){
{//Preparse block for help, config files, and outstream
PreParser pp=new PreParser(args, getClass(), false);
args=pp.args;
outstream=pp.outstream;
}
//Set shared static variables prior to parsing
ReadWrite.USE_PIGZ=ReadWrite.USE_UNPIGZ=true;
ReadWrite.setZipThreads(Shared.threads());
Shared.capBufferLen(50);
ReadWrite.ZIPLEVEL=9;
{//Parse the arguments
final Parser parser=parse(args);
Parser.processQuality();
maxReads=parser.maxReads;
overwrite=ReadStats.overwrite=parser.overwrite;
append=ReadStats.append=parser.append;
in1=parser.in1;
qfin1=parser.qfin1;
extin=parser.extin;
outPattern=parser.out1;
extout=parser.extout;
}
validateParams();
fixExtensions(); //Add or remove .gz or .bz2 as needed
checkFileExistence(); //Ensure files can be read and written
checkStatics(); //Adjust file-related static fields as needed for this program
//Create input FileFormat objects
ffin1=FileFormat.testInput(in1, FileFormat.FASTQ, extin, true, true);
numTypes=sequenceTypes.length;
readsOut=new long[numTypes];
basesOut=new long[numTypes];
consensusSequences=loadConsensusSequenceFromFile();
}
/*--------------------------------------------------------------*/
/*---------------- Initialization Helpers ----------------*/
/*--------------------------------------------------------------*/
/** Parse arguments from the command line */
private Parser parse(String[] args){
//Create a parser object
Parser parser=new Parser();
//Set any necessary Parser defaults here
//parser.foo=bar;
//Parse each argument
for(int i=0; i<args.length; i++){
String arg=args[i];
//Break arguments into their constituent parts, in the form of "a=b"
String[] split=arg.split("=");
String a=split[0].toLowerCase();
String b=split.length>1 ? split[1] : null;
if(b!=null && b.equalsIgnoreCase("null")){b=null;}
if(a.equals("verbose")){
verbose=Parse.parseBoolean(b);
}else if(a.equals("ordered")){
ordered=Parse.parseBoolean(b);
}else if(a.equalsIgnoreCase("minid")){
minID=Float.parseFloat(b);
}else if(a.equalsIgnoreCase("minid2") || a.equalsIgnoreCase("refineid")){
refineID=Float.parseFloat(b);
}else if(a.equals("out") || a.equals("pattern") || a.equals("outpattern")){
parser.out1=b;
}else if(a.equals("type") || a.equals("types")){
parseTypes(b);
}else if(a.equals("parse_flag_goes_here")){
long fake_variable=Parse.parseKMG(b);
//Set a variable here
}else if(parser.parse(arg, a, b)){//Parse standard flags in the parser
//do nothing
}else{
outstream.println("Unknown parameter "+args[i]);
assert(false) : "Unknown parameter "+args[i];
}
}
return parser;
}
private void parseTypes(String b){
sequenceTypes=null;
if(b==null){
assert(false) : "'types' flag requires a list of types, such as 'types=16S,18S'";
sequenceTypes=new String[] {"Other"};
}else{
String[] split=b.split(",");
sequenceTypes=new String[split.length+1];
sequenceTypes[0]="Other";
for(int i=0; i<split.length; i++){
String s=split[i].replace('s', 'S');
if(s.startsWith("its")){s=s.replaceFirst("its", "ITS");}
sequenceTypes[i+1]=s;
}
}
}
/** Add or remove .gz or .bz2 as needed */
private void fixExtensions(){
in1=Tools.fixExtension(in1);
qfin1=Tools.fixExtension(qfin1);
}
/** Ensure files can be read and written */
private void checkFileExistence(){
//Ensure input files can be read
if(!Tools.testInputFiles(false, true, in1)){
throw new RuntimeException("\nCan't read some input files.\n");
}
if(outPattern==null){return;}
if(!outPattern.contains("#")){
throw new RuntimeException("OutPattern must contain '#' symbol: "+outPattern);
}
for(String type : sequenceTypes) {
String out=outPattern.replaceFirst("#", type);
//Ensure output files can be written
if(!Tools.testOutputFiles(overwrite, append, false, out)){
outstream.println((outPattern==null)+", "+(out==null)+", "+outPattern+", "+out);
throw new RuntimeException("\n\noverwrite="+overwrite+"; Can't write to output file "+out+"\n");
}
//Ensure that no file was specified multiple times
if(!Tools.testForDuplicateFiles(true, in1, out)){
throw new RuntimeException("\nSome file names were specified multiple times.\n");
}
}
}
/** Adjust file-related static fields as needed for this program */
private static void checkStatics(){
//Adjust the number of threads for input file reading
if(!ByteFile.FORCE_MODE_BF1 && !ByteFile.FORCE_MODE_BF2 && Shared.threads()>2){
ByteFile.FORCE_MODE_BF2=true;
}
assert(FastaReadInputStream.settingsOK());
}
/** Ensure parameter ranges are within bounds and required parameters are set */
private boolean validateParams(){
// assert(minfoo>0 && minfoo<=maxfoo) : minfoo+", "+maxfoo;
if(in1==null){throw new RuntimeException("Error - at least one input file is required.");}
return true;
}
private final Read[][] loadConsensusSequenceFromFile(){
Read[][] seqs=new Read[numTypes][];
m16S_index=Tools.find("m16S", sequenceTypes);
m18S_index=Tools.find("m18S", sequenceTypes);
p16S_index=Tools.find("p16S", sequenceTypes);
boolean stripM16S=(m16S_index>=0);
boolean stripM18S=(m18S_index>=0);
boolean stripP16S=(p16S_index>=0);
for(int st=1; st<numTypes; st++){
String name=sequenceTypes[st];
boolean is16S=name.equalsIgnoreCase("16S");
boolean is18S=name.equalsIgnoreCase("18S");
seqs[st]=ProkObject.loadConsensusSequenceType(name, ((is16S && stripM16S) || (is18S && stripM18S)), (is16S && stripP16S));
}
return seqs;
}
/*--------------------------------------------------------------*/
/*---------------- Outer Methods ----------------*/
/*--------------------------------------------------------------*/
/** Create read streams and process all data */
void process(Timer t){
//Turn off read validation in the input threads to increase speed
final boolean vic=Read.VALIDATE_IN_CONSTRUCTOR;
Read.VALIDATE_IN_CONSTRUCTOR=Shared.threads()<4;
//Create a read input stream
final ConcurrentReadInputStream cris=makeCris();
//Optionally create a read output stream
final ConcurrentReadOutputStream[] rosa=makeCrosArray();
//Reset counters
readsProcessed=0;
basesProcessed=0;
Arrays.fill(readsOut, 0);
Arrays.fill(basesOut, 0);
//Process the reads in separate threads
spawnThreads(cris, rosa);
if(verbose){outstream.println("Finished; closing streams.");}
//Write anything that was accumulated by ReadStats
errorState|=ReadStats.writeAll();
//assert(!errorState);
//Close the read streams
errorState|=ReadWrite.closeStreams(cris, rosa);
//assert(!errorState);
//Reset read validation
Read.VALIDATE_IN_CONSTRUCTOR=vic;
long readsOut2=shared.Vector.sum(readsOut)-readsOut[0];
long basesOut2=shared.Vector.sum(basesOut)-basesOut[0];
//Report timing and results
t.stop();
outstream.println(Tools.timeReadsBasesProcessed(t, readsProcessed, basesProcessed, 8));
outstream.println(Tools.readsBasesOut(readsProcessed, basesProcessed, readsOut2, basesOut2, 8, true));
outstream.println();
outstream.println(Tools.string("Type", "Count", 8));
for(int type=0; type<numTypes; type++){
outstream.println(Tools.number(sequenceTypes[type], readsOut[type], 8));
}
//Throw an exception of there was an error in a thread
if(errorState){
throw new RuntimeException(getClass().getName()+" terminated in an error state; the output may be corrupt.");
}
}
private ConcurrentReadInputStream makeCris(){
ConcurrentReadInputStream cris=ConcurrentReadInputStream.getReadInputStream(maxReads, true, ffin1, null, qfin1, null);
cris.start(); //Start the stream
if(verbose){outstream.println("Started cris");}
return cris;
}
private ConcurrentReadOutputStream[] makeCrosArray(){
ConcurrentReadOutputStream[] rosa=new ConcurrentReadOutputStream[numTypes];
for(int i=0; i<numTypes; i++){
String type=sequenceTypes[i];
final ConcurrentReadOutputStream ros=makeCros(type);
rosa[i]=ros;
}
return rosa;
}
private ConcurrentReadOutputStream makeCros(String type){
if(outPattern==null){return null;}
//Select output buffer size based on whether it needs to be ordered
final int buff=(ordered ? Tools.mid(2, 16, (Shared.threads()*2)/3) : 4);
final String fname=outPattern.replaceFirst("#", type);
FileFormat ff=FileFormat.testOutput(fname, FileFormat.FASTA, extout, true, overwrite, append, ordered);
final ConcurrentReadOutputStream ros=ConcurrentReadOutputStream.getStream(ff, null, buff, null, false);
ros.start(); //Start the stream
return ros;
}
/*--------------------------------------------------------------*/
/*---------------- Thread Management ----------------*/
/*--------------------------------------------------------------*/
/** Spawn process threads */
private void spawnThreads(final ConcurrentReadInputStream cris, final ConcurrentReadOutputStream[] rosa){
//Do anything necessary prior to processing
//Determine how many threads may be used
final int threads=Shared.threads();
//Fill a list with ProcessThreads
ArrayList<ProcessThread> alpt=new ArrayList<ProcessThread>(threads);
for(int i=0; i<threads; i++){
alpt.add(new ProcessThread(cris, rosa, i));
}
//Start the threads and wait for them to finish
boolean success=ThreadWaiter.startAndWait(alpt, this);
errorState&=!success;
//assert(!errorState);
//Do anything necessary after processing
}
@Override
public final void accumulate(ProcessThread pt){
readsProcessed+=pt.readsProcessedT;
basesProcessed+=pt.basesProcessedT;
Tools.add(readsOut, pt.readsOutT);
Tools.add(basesOut, pt.basesOutT);
errorState|=(!pt.success);
//assert(!errorState);
}
@Override
public final boolean success(){return !errorState;}
/*--------------------------------------------------------------*/
/*---------------- Inner Methods ----------------*/
/*--------------------------------------------------------------*/
/*--------------------------------------------------------------*/
/*---------------- Inner Classes ----------------*/
/*--------------------------------------------------------------*/
/** This class is static to prevent accidental writing to shared variables.
* It is safe to remove the static modifier. */
class ProcessThread extends Thread {
//Constructor
ProcessThread(final ConcurrentReadInputStream cris_, final ConcurrentReadOutputStream[] rosa_, final int tid_){
cris=cris_;
rosa=rosa_;
tid=tid_;
}
//Called by start()
@Override
public void run(){
//Do anything necessary prior to processing
//Process the reads
processInner();
//Do anything necessary after processing
//Indicate successful exit status
success=true;
}
/** Iterate through the reads */
void processInner(){
//Grab the first ListNum of reads
ListNum<Read> ln=cris.nextList();
//Check to ensure pairing is as expected
if(ln!=null && !ln.isEmpty()){
Read r=ln.get(0);
assert(r.mate==null);
}
//As long as there is a nonempty read list...
while(ln!=null && ln.size()>0){
// if(verbose){outstream.println("Fetched "+reads.size()+" reads.");} //Disabled due to non-static access
processList(ln);
//Notify the input stream that the list was used
cris.returnList(ln);
// if(verbose){outstream.println("Returned a list.");} //Disabled due to non-static access
//Fetch a new list
ln=cris.nextList();
}
//Notify the input stream that the final list was used
if(ln!=null){
cris.returnList(ln.id, ln.list==null || ln.list.isEmpty());
}
}
void processList(ListNum<Read> ln){
//Grab the actual read list from the ListNum
final ArrayList<Read> reads=ln.list;
@SuppressWarnings("unchecked")
final ArrayList<Read>[] out=new ArrayList[numTypes];
for(int i=0; i<numTypes; i++){
ArrayList<Read> list=new ArrayList<Read>(50);
out[i]=list;
}
//Loop through each read in the list
for(int idx=0; idx<reads.size(); idx++){
final Read r1=reads.get(idx);
//Validate reads in worker threads
if(!r1.validated()){r1.validate(true);}
//Track the initial length for statistics
final int initialLength1=r1.length();
final int initialLength2=r1.mateLength();
//Increment counters
readsProcessedT+=r1.pairCount();
basesProcessedT+=initialLength1+initialLength2;
{
//Reads are processed in this block.
final int type=processRead(r1);
readsOutT[type]+=r1.pairCount();
basesOutT[type]+=r1.pairLength();
out[type].add(r1);
}
}
//Output reads to the output stream
if(rosa!=null){
for(int type=0; type<numTypes; type++){
rosa[type].add(out[type], ln.id);
}
}
}
/**
* Process a read.
* @param r1 Read 1
* @return The best-matching type, or 0 for no matches.
*/
private int processRead(final Read r){
int bestType=0;
float bestID=-1;
for(int type=1; type<numTypes; type++){//Align to only the overall consensus
Read[] refs=consensusSequences[type];
float id=align(r, refs, 0, 1);
if(id>bestID && id>=minID){
bestType=type;
bestID=id;
}
}
if(bestType<1 || bestID<refineID || bestType==p16S_index){//If nothing met minID, or if it matched chloro, align to clade-specific consensuses
for(int type=1; type<numTypes; type++){
Read[] refs=consensusSequences[type];
float id=align(r, refs, 1, refs.length);
if(id>bestID && id>=minID){
bestType=type;
bestID=id;
}
}
}
r.obj=bestID;//If desired... in actuality, more info might be useful, like alignment length
return bestID<minID ? 0 : bestType;
}
private float align(Read r, Read[] refs, int minRef, int maxRef){
float bestID=-1;
if(refs!=null){
for(int i=minRef; i<maxRef; i++){
Read ref=refs[i];
float id=align(r.bases, ref.bases);
bestID=Tools.max(id, bestID);
}
}
return bestID;
}
private float align(byte[] query, byte[] ref){
int a=0, b=ref.length-1;
int[] max=ssa.fillUnlimited(query, ref, a, b, -9999);
if(max==null){return 0;}
final int rows=max[0];
final int maxCol=max[1];
final int maxState=max[2];
final float id=ssa.tracebackIdentity(query, ref, a, b, rows, maxCol, maxState, null);
return id;
}
SingleStateAlignerFlat2 ssa=new SingleStateAlignerFlat2();
/** Number of reads processed by this thread */
protected long readsProcessedT=0;
/** Number of bases processed by this thread */
protected long basesProcessedT=0;
/** Number of reads retained by this thread */
protected long[] readsOutT=new long[numTypes];
/** Number of bases retained by this thread */
protected long[] basesOutT=new long[numTypes];
/** True only if this thread has completed successfully */
boolean success=false;
/** Shared input stream */
private final ConcurrentReadInputStream cris;
/** Shared output stream */
private final ConcurrentReadOutputStream[] rosa;
/** Thread ID */
final int tid;
}
/*--------------------------------------------------------------*/
/*---------------- Fields ----------------*/
/*--------------------------------------------------------------*/
/** Primary input file path */
private String in1=null;
private String qfin1=null;
/** Primary output file path */
private String outPattern=null;
/** Override input file extension */
private String extin=null;
/** Override output file extension */
private String extout=null;
float minID=0.59f; //This could be a per-type value
float refineID=0.70f; //Refine alignment if best is less than this
private int m16S_index=-2;
private int m18S_index=-2;
private int p16S_index=-2;
/*--------------------------------------------------------------*/
/** Number of reads processed */
protected long readsProcessed=0;
/** Number of bases processed */
protected long basesProcessed=0;
/** Quit after processing this many input reads; -1 means no limit */
private long maxReads=-1;
private String[] sequenceTypes=new String[] {"Other", "16S", "18S", "23S", "5S", "m16S", "m18S", "p16S"};
private final int numTypes;//=sequenceTypes.length;
final Read[][] consensusSequences;
/** Number of reads retained */
final long[] readsOut;
/** Number of bases retained */
final long[] basesOut;
/*--------------------------------------------------------------*/
/*---------------- Final Fields ----------------*/
/*--------------------------------------------------------------*/
/** Primary input file */
private final FileFormat ffin1;
@Override
public final ReadWriteLock rwlock() {return rwlock;}
private final ReadWriteLock rwlock=new ReentrantReadWriteLock();
/*--------------------------------------------------------------*/
/*---------------- Static Fields ----------------*/
/*--------------------------------------------------------------*/
/*--------------------------------------------------------------*/
/*---------------- Common Fields ----------------*/
/*--------------------------------------------------------------*/
/** Print status messages to this output stream */
private PrintStream outstream=System.err;
/** Print verbose messages */
public static boolean verbose=false;
/** True if an error was encountered */
public boolean errorState=false;
/** Overwrite existing output files */
private boolean overwrite=true;
/** Append to existing output files */
private boolean append=false;
/** Reads are output in input order */
private boolean ordered=true;
}
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