deacon 0.15.0

Accelerated DNA sequence search and [host] depletion using minimizers
Documentation
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
128
129
130
131
132
133
134
135
136
137
138
139
140
141
142
143
144
145
146
147
148
149
150
151
152
153
154
155
156
157
158
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
174
175
176
177
178
179
180
181
182
183
184
185
186
187
188
189
190
191
192
193
194
195
196
197
198
199
200
201
202
203
204
205
206
207
208
209
210
211
212
213
214
215
216
217
218
219
220
221
222
223
224
225
226
227
228
229
230
231
232
233
234
235
236
237
238
239
240
241
242
243
244
245
246
247
248
249
250
251
252
253
254
255
256
257
258
259
260
261
262
263
264
265
266
267
268
269
270
271
272
273
274
275
276
277
278
279
280
281
282
283
284
285
286
287
288
289
290
291
292
293
294
295
296
297
298
299
300
301
302
303
304
305
306
307
308
309
310
311
312
313
314
315
316
317
318
319
320
321
322
323
324
325
326
327
328
329
330
331
332
333
334
335
336
337
338
339
340
341
342
343
344
345
346
347
348
349
350
351
352
353
354
355
356
357
358
359
360
361
362
363
364
365
366
367
368
369
370
371
372
373
374
375
376
377
378
379
380
381
382
383
384
385
386
387
388
389
390
391
392
393
394
395
396
397
398
399
400
401
402
403
404
405
406
407
408
409
410
411
412
413
414
415
416
417
418
419
420
421
422
423
424
425
426
427
428
429
430
431
432
433
434
435
436
437
438
439
440
441
442
443
444
445
446
447
448
449
450
451
452
453
454
455
456
457
458
459
460
461
462
463
464
465
466
467
468
469
470
471
472
473
474
475
476
477
478
479
480
481
482
483
484
485
486
487
488
489
490
491
492
493
494
495
496
497
498
499
500
501
502
503
504
505
506
507
508
509
510
511
512
513
514
515
516
517
518
519
520
521
522
523
524
525
526
527
528
529
530
531
532
533
534
535
536
537
538
539
540
541
542
543
544
545
546
547
548
549
550
551
552
553
554
555
556
557
558
559
560
561
562
563
564
565
566
567
568
569
570
571
572
573
574
575
576
577
578
579
580
581
582
583
584
585
586
587
588
589
590
591
592
593
594
595
596
597
598
599
600
601
602
603
604
605
606
607
608
609
610
611
612
613
614
615
616
617
618
619
620
621
622
623
624
625
626
627
628
629
630
631
632
633
634
635
636
637
638
639
640
641
642
643
644
645
646
647
648
649
650
651
652
653
654
655
656
657
658
659
660
661
662
663
664
665
666
667
668
669
670
671
672
673
674
675
676
677
678
679
680
681
682
683
684
685
686
687
688
689
690
691
692
693
694
695
696
697
698
699
700
701
702
703
704
705
706
707
708
709
710
711
712
713
714
715
716
717
718
719
720
721
722
723
724
725
726
727
728
729
730
731
732
733
734
735
736
737
738
739
740
741
742
743
744
745
746
747
748
749
750
751
752
753
754
755
756
757
758
759
760
761
762
763
764
765
766
767
768
769
770
771
772
773
774
775
776
777
778
779
780
781
782
783
784
785
786
787
788
789
790
791
792
793
794
795
796
797
798
799
800
801
802
803
804
805
806
807
808
809
810
811
812
813
814
815
816
817
818
819
820
821
822
823
824
825
826
827
828
829
830
831
832
833
834
835
836
837
838
839
840
841
842
843
844
845
846
847
848
849
850
851
852
853
854
855
856
857
858
859
860
861
862
863
864
865
866
867
868
869
870
871
872
873
874
875
876
877
878
879
880
881
882
883
884
885
886
887
888
889
890
891
892
893
894
895
896
897
898
899
900
901
902
903
904
905
906
907
908
909
910
911
912
913
914
915
916
917
918
919
920
921
922
923
924
925
926
927
928
929
930
931
932
933
934
935
936
937
938
939
940
941
942
943
944
945
946
947
948
949
950
951
952
953
954
955
956
957
958
959
960
961
962
963
964
965
966
967
968
969
970
971
972
973
974
975
976
977
978
979
980
981
982
983
984
985
986
987
988
989
990
991
992
993
994
995
996
997
998
999
1000
1001
1002
1003
1004
1005
1006
1007
1008
1009
1010
1011
1012
1013
1014
1015
1016
1017
1018
1019
1020
1021
1022
1023
1024
1025
1026
1027
1028
1029
1030
1031
1032
1033
1034
1035
1036
1037
1038
1039
1040
1041
1042
1043
1044
1045
1046
1047
1048
1049
1050
1051
1052
1053
1054
1055
1056
1057
1058
1059
1060
1061
1062
1063
1064
1065
1066
1067
1068
1069
1070
1071
1072
1073
1074
1075
1076
1077
1078
1079
1080
1081
1082
1083
1084
1085
1086
1087
1088
1089
1090
1091
1092
1093
1094
1095
1096
1097
1098
1099
1100
1101
1102
1103
1104
1105
1106
1107
1108
1109
1110
1111
1112
1113
1114
1115
1116
1117
1118
1119
use crate::filter::{Buffers, ProcessingStats};
use crate::minimizers::KmerHasher;
use crate::{FixedRapidHasher, IndexConfig, RapidHashSet};
use anyhow::{Context, Result};
use bincode::serde::{decode_from_std_read, encode_into_std_write};
use paraseq::Record;
use paraseq::prelude::{ParallelProcessor, ParallelReader};
use parking_lot::Mutex;
use serde::{Deserialize, Serialize};
use std::fs::File;
use std::io::{self, BufReader, BufWriter, Read, Write};
use std::path::{Path, PathBuf};
use std::sync::{Arc, OnceLock};
use std::time::Instant;

#[cfg(feature = "fetch")]
use indicatif::ProgressBar;

/// Index format version
pub const INDEX_FORMAT_VERSION: u8 = 3;

/// Serialisable header for the index file
#[derive(Serialize, Deserialize, Debug)]
pub struct IndexHeader {
    pub format_version: u8,
    pub kmer_length: u8,
    pub window_size: u8,
}

impl IndexHeader {
    pub fn new(kmer_length: u8, window_size: u8) -> Self {
        IndexHeader {
            format_version: INDEX_FORMAT_VERSION,
            kmer_length,
            window_size,
        }
    }

    /// Validate header
    pub fn validate(&self) -> anyhow::Result<()> {
        if self.format_version != INDEX_FORMAT_VERSION {
            return Err(anyhow::anyhow!(
                "Unsupported index format version: {}",
                self.format_version
            ));
        }

        let k = self.kmer_length as usize;
        let w = self.window_size as usize;

        // Check constraints: k <= 61, k+w <= 96, k+w even (ensures k odd and k+w-1 odd)
        if k > 61 || k + w > 96 || (k + w) % 2 != 0 {
            return Err(anyhow::anyhow!(
                "Invalid k-w combination: k={}, w={}, k+w={} (constraints: k<=61, k+w<=96, k+w even)",
                k,
                w,
                k + w
            ));
        }

        Ok(())
    }

    /// Get k
    pub fn kmer_length(&self) -> u8 {
        self.kmer_length
    }

    /// Get w
    pub fn window_size(&self) -> u8 {
        self.window_size
    }
}

/// Load just the header and count from an index file
pub fn load_header_and_count<P: AsRef<Path>>(path: &P) -> Result<(IndexHeader, usize)> {
    let file =
        File::open(path).context(format!("Failed to open index file {:?}", path.as_ref()))?;
    let mut reader = BufReader::new(file);

    let config = bincode::config::standard().with_fixed_int_encoding();

    // Deserialise header
    let header: IndexHeader =
        decode_from_std_read(&mut reader, config).context("Failed to deserialise index header")?;
    header.validate()?;

    // Deserialise the count of minimizers
    let count: usize = decode_from_std_read(&mut reader, config)
        .context("Failed to deserialise minimizer count")?;

    Ok((header, count))
}

static INDEX: OnceLock<(PathBuf, crate::MinimizerSet, IndexHeader)> = OnceLock::new();

pub fn load_minimizers_cached(
    path: &Path,
) -> Result<(&'static crate::MinimizerSet, &'static IndexHeader)> {
    let (p, minimizers, header) = INDEX.get_or_init(|| {
        let (m, h) = load_minimizers(path).unwrap();
        (path.to_owned(), m, h)
    });
    assert_eq!(
        p, path,
        "Currently, the server can only have one index loaded."
    );

    Ok((minimizers, header))
}

/// Load minimizers from an index file
pub fn load_minimizers(path: &Path) -> Result<(crate::MinimizerSet, IndexHeader)> {
    let file = File::open(path).context(format!("Failed to open index file {:?}", path))?;
    let mut reader = BufReader::with_capacity(1 << 20, file);
    let config = bincode::config::standard().with_fixed_int_encoding();

    // Deserialise header
    let header: IndexHeader =
        decode_from_std_read(&mut reader, config).context("Failed to deserialise index header")?;
    header.validate()?;

    // Deserialise the count of minimizers
    let count: usize = decode_from_std_read(&mut reader, config)
        .context("Failed to deserialise minimizer count")?;

    let bytes_per_minimizer = (header.kmer_length as usize).div_ceil(4);

    let minimizers = if header.kmer_length <= 32 {
        // Read as u64 with packed byte-aligned format
        let mut set = RapidHashSet::<u64>::with_capacity_and_hasher(count, FixedRapidHasher);
        const B: usize = 16 * 1024;
        let mut buffer = vec![0u8; bytes_per_minimizer * B];
        for i in (0..count).step_by(B) {
            let batch_count = B.min(count - i);
            let batch_bytes = bytes_per_minimizer * batch_count;
            let batch = &mut buffer[..batch_bytes];
            reader.read_exact(batch).context(format!(
                "Failed to load minimizer batch {}, index may be corrupt",
                i / B
            ))?;

            // Extract minimizers from packed bytes
            for j in 0..batch_count {
                let start = j * bytes_per_minimizer;
                let end = start + bytes_per_minimizer;
                let mut minimizer_bytes = [0u8; 8];
                minimizer_bytes[..bytes_per_minimizer].copy_from_slice(&batch[start..end]);
                set.insert(u64::from_le_bytes(minimizer_bytes));
            }
        }
        crate::MinimizerSet::U64(set)
    } else {
        // Read as u128 with packed byte-aligned format
        let mut set = RapidHashSet::<u128>::with_capacity_and_hasher(count, FixedRapidHasher);
        const B: usize = 16 * 1024;
        let mut buffer = vec![0u8; bytes_per_minimizer * B];
        for i in (0..count).step_by(B) {
            let batch_count = B.min(count - i);
            let batch_bytes = bytes_per_minimizer * batch_count;
            let batch = &mut buffer[..batch_bytes];
            reader.read_exact(batch).context(format!(
                "Failed to load minimizer batch {}, index may be corrupt",
                i / B
            ))?;

            // Extract minimizers from packed bytes
            for j in 0..batch_count {
                let start = j * bytes_per_minimizer;
                let end = start + bytes_per_minimizer;
                let mut minimizer_bytes = [0u8; 16];
                minimizer_bytes[..bytes_per_minimizer].copy_from_slice(&batch[start..end]);
                set.insert(u128::from_le_bytes(minimizer_bytes));
            }
        }
        crate::MinimizerSet::U128(set)
    };

    // Validate that we loaded the expected number of minimizers
    let loaded_count = minimizers.len();
    if loaded_count != count {
        return Err(anyhow::anyhow!(
            "Failed to load expected number of minimizers; expected {} and observed {}. Index may be corrupt",
            count,
            loaded_count
        ));
    }

    Ok((minimizers, header))
}

/// Helper function to write minimizers to output file or stdout
pub fn dump_minimizers(
    minimizers: &crate::MinimizerSet,
    header: &IndexHeader,
    output_path: Option<&Path>,
) -> Result<()> {
    // Create writer based on output path
    let mut writer: BufWriter<Box<dyn Write>> = match output_path {
        Some(path) if path.as_os_str() != "-" => BufWriter::new(Box::new(
            File::create(path).context("Failed to create output file")?,
        )),
        _ => BufWriter::new(Box::new(io::stdout())),
    };

    // Use fixed-width little-endian encoding for all values.
    let config = bincode::config::standard().with_fixed_int_encoding();
    encode_into_std_write(header, &mut writer, config)
        .context("Failed to serialise index header")?;

    // Serialise the count of minimizers first
    let count = minimizers.len();
    encode_into_std_write(count, &mut writer, config)
        .context("Failed to serialise minimizer count")?;

    // Serialise minimizers in byte-aligned packed format
    let bytes_per_minimizer = (header.kmer_length as usize).div_ceil(4);
    match minimizers {
        crate::MinimizerSet::U64(set) => {
            for &val in set {
                // Write only the required bytes (little-endian)
                let bytes = val.to_le_bytes();
                writer
                    .write_all(&bytes[..bytes_per_minimizer])
                    .context("Failed to write minimizer")?;
            }
        }
        crate::MinimizerSet::U128(set) => {
            for &val in set {
                // Write only the required bytes (little-endian)
                let bytes = val.to_le_bytes();
                writer
                    .write_all(&bytes[..bytes_per_minimizer])
                    .context("Failed to write minimizer")?;
            }
        }
    }
    Ok(())
}

/// Dump indexed minimizers to FASTA
pub fn dump(index_path: &Path, output_path: Option<&Path>) -> Result<()> {
    // Load the index
    let (minimizers, header) = load_minimizers(index_path).context("Failed to load index")?;

    // Create writer based on output path
    let mut writer: BufWriter<Box<dyn Write>> = match output_path {
        Some(path) if path.as_os_str() != "-" => BufWriter::new(Box::new(
            File::create(path).context("Failed to create output file")?,
        )),
        _ => BufWriter::new(Box::new(io::stdout())),
    };

    // Write FASTA
    let mut counter = 0;
    match minimizers {
        crate::MinimizerSet::U64(set) => {
            for &minimizer in &set {
                counter += 1;
                let sequence = crate::minimizers::decode_u64(minimizer, header.kmer_length);
                writeln!(writer, ">{}", counter)?;
                writeln!(writer, "{}", String::from_utf8_lossy(&sequence))?;
            }
        }
        crate::MinimizerSet::U128(set) => {
            for &minimizer in &set {
                counter += 1;
                let sequence = crate::minimizers::decode_u128(minimizer, header.kmer_length);
                writeln!(writer, ">{}", counter)?;
                writeln!(writer, "{}", String::from_utf8_lossy(&sequence))?;
            }
        }
    }

    writer.flush()?;
    Ok(())
}

fn reader_with_inferred_batch_size(
    in_path: Option<&Path>,
) -> Result<paraseq::fastx::Reader<Box<dyn Read + Send>>> {
    let mut reader = paraseq::fastx::Reader::from_optional_path(in_path).unwrap();
    reader.update_batch_size_in_bp(256 * 1024)?;
    Ok(reader)
}

#[derive(Clone)]
struct BuildIndexProcessor<'c> {
    config: &'c IndexConfig,
    hasher: KmerHasher,
    // Local buffers
    buffers: Buffers,
    local_stats: ProcessingStats,
    local_minimizers_u64: Option<RapidHashSet<u64>>,
    local_minimizers_u128: Option<RapidHashSet<u128>>,
    local_record_info: Vec<(Vec<u8>, usize)>, // (record_id, seq_len) for progress output
    // Global state
    global_stats: Arc<Mutex<ProcessingStats>>,
    global_minimizers_u64: Arc<Mutex<Option<RapidHashSet<u64>>>>,
    global_minimizers_u128: Arc<Mutex<Option<RapidHashSet<u128>>>>,
}

impl<Rf: Record> ParallelProcessor<Rf> for BuildIndexProcessor<'_> {
    fn process_record(&mut self, record: Rf) -> paraseq::parallel::Result<()> {
        let seq = record.seq();
        self.local_stats.total_seqs += 1;
        self.local_stats.total_bp += seq.len() as u64;

        crate::minimizers::fill_minimizers(
            &seq,
            &self.hasher,
            self.config.kmer_length,
            self.config.window_size,
            self.config.entropy_threshold,
            &mut self.buffers,
        );

        // Extend appropriate local set based on type
        match &mut self.buffers.minimizers {
            crate::MinimizerVec::U64(vec) => {
                self.local_minimizers_u64
                    .as_mut()
                    .unwrap()
                    .extend(vec.iter());
            }
            crate::MinimizerVec::U128(vec) => {
                self.local_minimizers_u128
                    .as_mut()
                    .unwrap()
                    .extend(vec.iter());
            }
        }

        // Store record info for progress output (printed sequentially in on_batch_complete)
        if !self.config.quiet {
            self.local_record_info
                .push((record.id().to_vec(), seq.len()));
        }

        Ok(())
    }

    fn on_batch_complete(&mut self) -> paraseq::parallel::Result<()> {
        // Merge local minimizers into global set and get new total count
        let minimizer_count = if let Some(local) = &mut self.local_minimizers_u64 {
            let mut global = self.global_minimizers_u64.lock();
            global.as_mut().unwrap().extend(local.iter());
            local.clear();
            global.as_ref().unwrap().len()
        } else {
            let mut global = self.global_minimizers_u128.lock();
            global
                .as_mut()
                .unwrap()
                .extend(self.local_minimizers_u128.as_ref().unwrap().iter());
            self.local_minimizers_u128.as_mut().unwrap().clear();
            global.as_ref().unwrap().len()
        };

        // Update global stats
        {
            let mut stats = self.global_stats.lock();
            stats.total_seqs += self.local_stats.total_seqs;
            stats.total_bp += self.local_stats.total_bp;
            self.local_stats = ProcessingStats::default();
        }

        // Print progress for each record in this batch (sequentially, after merging)
        if !self.config.quiet {
            for (record_id, seq_len) in &self.local_record_info {
                let id_str = std::str::from_utf8(record_id).unwrap_or("unknown");
                eprintln!(
                    "  {} ({}bp), total minimizers: {}",
                    id_str, seq_len, minimizer_count
                );
            }
            self.local_record_info.clear();
        }

        Ok(())
    }
}

/// Build an index of minimizers from a fastx file
pub fn build(config: &IndexConfig) -> Result<()> {
    let start_time = Instant::now();
    let path = &config.input_path;

    let version: String = env!("CARGO_PKG_VERSION").to_string();

    // Build options string similar to filter
    let mut options = Vec::<String>::new();
    if config.threads > 0 {
        options.push(format!("threads={}", config.threads));
    }

    eprintln!(
        "Deacon v{}; mode: build; input: single; options: {}",
        version,
        options.join(", ")
    );

    // Validate k-mer and window size constraints
    config.validate()?;

    let in_path = if path.as_os_str() == "-" {
        None
    } else {
        Some(path.as_path())
    };
    let reader = reader_with_inferred_batch_size(in_path)?;

    eprintln!(
        "Building index (k={}, w={})",
        config.kmer_length, config.window_size
    );

    let mut processor = if config.kmer_length <= 32 {
        BuildIndexProcessor {
            config,
            hasher: KmerHasher::new(config.kmer_length as usize),
            local_stats: ProcessingStats::default(),
            buffers: Buffers::new_u64(),
            local_minimizers_u64: Some(RapidHashSet::default()),
            local_minimizers_u128: None,
            local_record_info: Vec::new(),
            global_stats: Arc::new(Mutex::new(ProcessingStats::default())),
            global_minimizers_u64: Arc::new(Mutex::new(Some(RapidHashSet::default()))),
            global_minimizers_u128: Arc::new(Mutex::new(None)),
        }
    } else {
        BuildIndexProcessor {
            config,
            hasher: KmerHasher::new(config.kmer_length as usize),
            local_stats: ProcessingStats::default(),
            buffers: Buffers::new_u128(),
            local_minimizers_u64: None,
            local_minimizers_u128: Some(RapidHashSet::default()),
            local_record_info: Vec::new(),
            global_stats: Arc::new(Mutex::new(ProcessingStats::default())),
            global_minimizers_u64: Arc::new(Mutex::new(None)),
            global_minimizers_u128: Arc::new(Mutex::new(Some(RapidHashSet::default()))),
        }
    };
    reader.process_parallel(&mut processor, config.threads as usize)?;

    let all_minimizers = if config.kmer_length <= 32 {
        let set = Arc::try_unwrap(processor.global_minimizers_u64)
            .unwrap()
            .into_inner()
            .unwrap();
        crate::MinimizerSet::U64(set)
    } else {
        let set = Arc::try_unwrap(processor.global_minimizers_u128)
            .unwrap()
            .into_inner()
            .unwrap();
        crate::MinimizerSet::U128(set)
    };
    let stats = Arc::try_unwrap(processor.global_stats)
        .unwrap()
        .into_inner();

    eprintln!(
        "Indexed {} minimizers from {} record(s) ({}bp)",
        all_minimizers.len(),
        stats.total_seqs,
        stats.total_bp
    );

    let header = IndexHeader::new(config.kmer_length, config.window_size);

    // Write to output path or stdout
    dump_minimizers(&all_minimizers, &header, config.output_path.as_deref())?;

    let total_time = start_time.elapsed();
    eprintln!("Completed in {:.2?}", total_time);

    Ok(())
}

#[derive(Clone)]
struct DiffIndexProcessor {
    kmer_length: u8,
    window_size: u8,
    hasher: KmerHasher,
    // Local buffers
    buffers: Buffers,
    local_stats: ProcessingStats,
    local_minimizers_u64: Option<RapidHashSet<u64>>,
    local_minimizers_u128: Option<RapidHashSet<u128>>,
    // Global state
    global_stats: Arc<Mutex<ProcessingStats>>,
    initial_size: usize,
    global_minimizers_u64: Arc<Mutex<Option<RapidHashSet<u64>>>>,
    global_minimizers_u128: Arc<Mutex<Option<RapidHashSet<u128>>>>,
}

impl<Rf: Record> ParallelProcessor<Rf> for DiffIndexProcessor {
    fn process_record(&mut self, record: Rf) -> paraseq::parallel::Result<()> {
        let seq = record.seq();
        self.local_stats.total_seqs += 1;
        self.local_stats.total_bp += seq.len() as u64;

        crate::minimizers::fill_minimizers(
            &seq,
            &self.hasher,
            self.kmer_length,
            self.window_size,
            0.0,
            &mut self.buffers,
        );

        // Extend appropriate local set based on type
        match &mut self.buffers.minimizers {
            crate::MinimizerVec::U64(vec) => {
                self.local_minimizers_u64
                    .as_mut()
                    .unwrap()
                    .extend(vec.iter());
            }
            crate::MinimizerVec::U128(vec) => {
                self.local_minimizers_u128
                    .as_mut()
                    .unwrap()
                    .extend(vec.iter());
            }
        }

        Ok(())
    }

    fn on_batch_complete(&mut self) -> paraseq::parallel::Result<()> {
        // Write buffer to output
        let len = if let Some(local) = &mut self.local_minimizers_u64 {
            let mut global = self.global_minimizers_u64.lock();
            let global_set = global.as_mut().unwrap();
            for &minimizer in local.iter() {
                global_set.remove(&minimizer);
            }
            let len = global_set.len();
            local.clear();
            len
        } else {
            let mut global = self.global_minimizers_u128.lock();
            let global_set = global.as_mut().unwrap();
            for &minimizer in self.local_minimizers_u128.as_ref().unwrap().iter() {
                global_set.remove(&minimizer);
            }
            let len = global_set.len();
            self.local_minimizers_u128.as_mut().unwrap().clear();
            len
        };

        // Update global stats
        {
            let mut stats = self.global_stats.lock();
            stats.total_seqs += self.local_stats.total_seqs;
            stats.total_bp += self.local_stats.total_bp;

            let current_gb = stats.total_bp / 1_000_000_000;
            if current_gb > stats.last_reported {
                eprintln!(
                    "  Processed {} sequences ({}bp), removed {} minimizers",
                    stats.total_seqs,
                    stats.total_bp,
                    self.initial_size - len
                );
                stats.last_reported = current_gb;
            }

            self.local_stats = ProcessingStats::default();
        }

        Ok(())
    }
}

/// Stream minimizers from a FASTX file or stdin and remove those present in first_minimizers
fn stream_diff_fastx(
    fastx_path: &Path,
    kmer_length: u8,
    window_size: u8,
    first_header: &IndexHeader,
    threads: u16,
    first_minimizers: &mut crate::MinimizerSet,
) -> Result<(usize, usize)> {
    let path = fastx_path;

    // Validate k-mer and window size constraints
    let temp_config = crate::IndexConfig {
        input_path: PathBuf::new(),
        kmer_length,
        window_size,
        output_path: None,
        threads: 0,
        quiet: false,
        entropy_threshold: 0.0,
    };
    temp_config.validate()?;

    // Validate parameters match the first index
    if kmer_length != first_header.kmer_length() || window_size != first_header.window_size() {
        return Err(anyhow::anyhow!(
            "FASTX parameters (k={}, w={}) must match first index (k={}, w={})",
            kmer_length,
            window_size,
            first_header.kmer_length(),
            first_header.window_size()
        ));
    }

    if path.as_os_str() == "-" {
        eprintln!(
            "Second index: processing FASTX from stdin (k={}, w={})…",
            kmer_length, window_size
        );
    } else {
        eprintln!(
            "Second index: processing FASTX from file (k={}, w={})…",
            kmer_length, window_size
        );
    }

    let in_path = if path.as_os_str() == "-" {
        None
    } else {
        Some(path)
    };

    let reader = reader_with_inferred_batch_size(in_path)?;

    // Move first_minimizers into Arc<Mutex<>> for parallel access
    let initial_size = first_minimizers.len();
    let (mut processor, global_minimizers_u64, global_minimizers_u128) = match first_minimizers {
        crate::MinimizerSet::U64(set) => {
            let moved_set = std::mem::take(set);
            let arc = Arc::new(Mutex::new(Some(moved_set)));
            (
                DiffIndexProcessor {
                    kmer_length,
                    window_size,
                    hasher: KmerHasher::new(kmer_length as usize),
                    local_stats: ProcessingStats::default(),
                    buffers: Buffers::new_u64(),
                    local_minimizers_u64: Some(RapidHashSet::default()),
                    local_minimizers_u128: None,
                    global_stats: Arc::new(Mutex::new(ProcessingStats::default())),
                    initial_size,
                    global_minimizers_u64: arc.clone(),
                    global_minimizers_u128: Arc::new(Mutex::new(None)),
                },
                Some(arc),
                None,
            )
        }
        crate::MinimizerSet::U128(set) => {
            let moved_set = std::mem::take(set);
            let arc = Arc::new(Mutex::new(Some(moved_set)));
            (
                DiffIndexProcessor {
                    kmer_length,
                    window_size,
                    hasher: KmerHasher::new(kmer_length as usize),
                    local_stats: ProcessingStats::default(),
                    buffers: Buffers::new_u128(),
                    local_minimizers_u64: None,
                    local_minimizers_u128: Some(RapidHashSet::default()),
                    global_stats: Arc::new(Mutex::new(ProcessingStats::default())),
                    initial_size,
                    global_minimizers_u64: Arc::new(Mutex::new(None)),
                    global_minimizers_u128: arc.clone(),
                },
                None,
                Some(arc),
            )
        }
    };

    reader.process_parallel(&mut processor, threads as usize)?;

    // Extract results from Arc<Mutex<>> after processing completes
    let stats = processor.global_stats.lock().clone();
    *first_minimizers = if let Some(arc) = global_minimizers_u64 {
        let set = arc.lock().take().unwrap();
        crate::MinimizerSet::U64(set)
    } else {
        let set = global_minimizers_u128.unwrap().lock().take().unwrap();
        crate::MinimizerSet::U128(set)
    };

    eprintln!(
        "Processed {} sequences ({}bp) from FASTX file",
        stats.total_seqs, stats.total_bp
    );

    Ok((stats.total_seqs as usize, stats.total_bp as usize))
}

/// Compute the set difference between two minimizer indexes (A - B)
pub fn diff(
    first: &Path,
    second: &Path,
    kmer_length: Option<u8>,
    window_size: Option<u8>,
    threads: u16,
    output: Option<&Path>,
) -> Result<()> {
    let start_time = Instant::now();

    // Load first file (always an index)
    let (mut first_minimizers, header) = load_minimizers(first)?;
    eprintln!("First index: loaded {} minimizers", first_minimizers.len());

    // Guess if second file is an index or FASTX file
    let second_minimizers = if let (Some(k), Some(w)) = (kmer_length, window_size) {
        // Second file is a FASTX file - stream diff with provided k, w
        let before_count = first_minimizers.len();
        let (_seq_count, _total_bp) =
            stream_diff_fastx(second, k, w, &header, threads, &mut first_minimizers)?;

        // Report results
        eprintln!(
            "Removed {} minimizers, {} remaining",
            before_count - first_minimizers.len(),
            first_minimizers.len()
        );

        dump_minimizers(&first_minimizers, &header, output)?;

        let total_time = start_time.elapsed();
        eprintln!("Completed difference operation in {:.2?}", total_time);

        return Ok(());
    } else {
        // Try to load as index file first
        if let Ok((second_minimizers, second_header)) = load_minimizers(&second) {
            // Second file is an index file
            eprintln!(
                "Second index: loaded {} minimizers",
                second_minimizers.len()
            );

            // Validate compatible headers
            if second_header.kmer_length() != header.kmer_length()
                || second_header.window_size() != header.window_size()
            {
                return Err(anyhow::anyhow!(
                    "Incompatible headers: second index has k={}, w={}, but first index has k={}, w={}",
                    second_header.kmer_length(),
                    second_header.window_size(),
                    header.kmer_length(),
                    header.window_size()
                ));
            }

            second_minimizers
        } else {
            // Second file is not a valid index, treat as FASTX file
            // Use k and w from first index header and do a streaming diff
            let k = header.kmer_length();
            let w = header.window_size();

            // Count minimizers before diff
            let before_count = first_minimizers.len();

            let (_seq_count, _total_bp) =
                stream_diff_fastx(&second, k, w, &header, threads, &mut first_minimizers)?;

            // Report results
            eprintln!(
                "Removed {} minimizers, {} remaining",
                before_count - first_minimizers.len(),
                first_minimizers.len()
            );

            dump_minimizers(&first_minimizers, &header, output)?;

            let total_time = start_time.elapsed();
            eprintln!("Completed difference operation in {:.2?}", total_time);

            return Ok(());
        }
    };

    // Handle straightforward index-to-index diffing
    // Count minimizers before diff
    let before_count = first_minimizers.len();

    // Remove all minimizers in second_minimizers from first_minimizers
    first_minimizers.remove_all(&second_minimizers);

    // Report results
    eprintln!(
        "Removed {} minimizers, {} remaining",
        before_count - first_minimizers.len(),
        first_minimizers.len()
    );

    dump_minimizers(&first_minimizers, &header, output)?;

    let total_time = start_time.elapsed();
    eprintln!("Completed diff operation in {:.2?}", total_time);

    Ok(())
}

/// Show info about an index
pub fn info(index_path: &Path) -> Result<()> {
    let start_time = Instant::now();

    // Load index file
    let (minimizers, header) = load_minimizers(index_path)?;

    // Show index info
    eprintln!("Index information:");
    eprintln!("  Format version: {}", header.format_version);
    eprintln!("  K-mer length (k): {}", header.kmer_length());
    eprintln!("  Window size (w): {}", header.window_size());
    eprintln!("  Distinct minimizer count: {}", minimizers.len());

    let total_time = start_time.elapsed();
    eprintln!("Loaded index info in {:.2?}", total_time);

    Ok(())
}

/// Combine minimizer indexes (set union)
pub fn union(inputs: &[PathBuf], output: Option<&Path>) -> Result<()> {
    let start_time = Instant::now();
    // Check input files
    if inputs.is_empty() {
        return Err(anyhow::anyhow!(
            "No input files provided for union operation"
        ));
    }

    // Read all headers first to determine total capacity needed
    let mut headers_and_counts = Vec::new();

    for path in inputs {
        let (header, count) = load_header_and_count(path)?;
        headers_and_counts.push((header, count));
    }

    // Get header from first file for output
    let header = &headers_and_counts[0].0;

    eprintln!(
        "Combining indexes (k={}, w={})",
        header.kmer_length(),
        header.window_size()
    );

    // Verify all headers are compatible
    for (i, (file_header, _)) in headers_and_counts.iter().enumerate() {
        if file_header.kmer_length() != header.kmer_length()
            || file_header.window_size() != header.window_size()
        {
            return Err(anyhow::anyhow!(
                "Incompatible headers: index {} has k={}, w={}, but first index has k={}, w={}",
                i,
                file_header.kmer_length(),
                file_header.window_size(),
                header.kmer_length(),
                header.window_size()
            ));
        }
    }

    // Load first index to determine type (u64 vs u128)
    let (mut all_minimizers, _) = load_minimizers(&inputs[0])?;
    eprintln!("Index 1: loaded {} minimizers", all_minimizers.len());

    // Now load and merge remaining indexes
    for (i, path) in inputs.iter().enumerate().skip(1) {
        let (minimizers, _) = load_minimizers(path)?;
        let before_count = all_minimizers.len();

        // Merge minimizers (set union)
        all_minimizers.extend(minimizers);

        let expected_count = headers_and_counts[i].1;
        eprintln!(
            "Index {}: {} minimizers, added {}, total: {}",
            i + 1,
            expected_count,
            all_minimizers.len() - before_count,
            all_minimizers.len()
        );
    }

    dump_minimizers(&all_minimizers, header, output)?;

    let total_time = start_time.elapsed();
    eprintln!(
        "United {} indexes with {} total minimizers in {:.2?}",
        inputs.len(),
        all_minimizers.len(),
        total_time
    );

    Ok(())
}

pub fn intersect(inputs: &[PathBuf], output: Option<&Path>) -> Result<()> {
    let start_time = Instant::now();
    // Check inputs
    if inputs.len() < 2 {
        return Err(anyhow::anyhow!(
            "At least two input files are required for intersection operation"
        ));
    }

    // Read all headers first
    let mut headers_and_counts = Vec::new();

    for path in inputs {
        let (header, count) = load_header_and_count(path)?;
        headers_and_counts.push((header, count));
    }

    // Get header from first file for output
    let header = &headers_and_counts[0].0;

    eprintln!(
        "Intersecting indexes (k={}, w={})",
        header.kmer_length(),
        header.window_size()
    );

    // Check header compat
    for (i, (file_header, _)) in headers_and_counts.iter().enumerate() {
        if file_header.kmer_length() != header.kmer_length()
            || file_header.window_size() != header.window_size()
        {
            return Err(anyhow::anyhow!(
                "Incompatible headers: index {} has k={}, w={}, but first index has k={}, w={}",
                i,
                file_header.kmer_length(),
                file_header.window_size(),
                header.kmer_length(),
                header.window_size()
            ));
        }
    }

    // Load first index
    let (mut result_minimizers, _) = load_minimizers(&inputs[0])?;
    eprintln!("Index 1: loaded {} minimizers", result_minimizers.len());

    // Intersect with remaining indexes
    for (i, path) in inputs.iter().enumerate().skip(1) {
        let (minimizers, _) = load_minimizers(path)?;

        // Intersect minimizers (set intersection)
        result_minimizers.intersect(&minimizers);

        let expected_count = headers_and_counts[i].1;
        eprintln!(
            "Index {}: {} minimizers, retained {}, total: {}",
            i + 1,
            expected_count,
            result_minimizers.len(),
            result_minimizers.len()
        );
    }

    dump_minimizers(&result_minimizers, header, output)?;

    let total_time = start_time.elapsed();
    eprintln!(
        "Intersected {} indexes with {} common minimizers in {:.2?}",
        inputs.len(),
        result_minimizers.len(),
        total_time
    );

    Ok(())
}

#[cfg(test)]
mod tests {
    use super::*;

    #[test]
    fn test_header_creation() {
        let header = IndexHeader::new(31, 21);

        assert_eq!(header.format_version, 3);
        assert_eq!(header.kmer_length(), 31);
        assert_eq!(header.window_size(), 21);
    }

    #[test]
    fn test_header_validation() {
        // Valid header (v3 only)
        let valid_header_v3 = IndexHeader {
            format_version: 3,
            kmer_length: 31,
            window_size: 21,
        };
        assert!(valid_header_v3.validate().is_ok());

        // Invalid format versions
        let invalid_header_v1 = IndexHeader {
            format_version: 1,
            kmer_length: 31,
            window_size: 21,
        };
        assert!(invalid_header_v1.validate().is_err());

        let invalid_header_v2 = IndexHeader {
            format_version: 2,
            kmer_length: 31,
            window_size: 21,
        };
        assert!(invalid_header_v2.validate().is_err());

        let invalid_header_v4 = IndexHeader {
            format_version: 4,
            kmer_length: 31,
            window_size: 21,
        };
        assert!(invalid_header_v4.validate().is_err());

        // Test k <= 61 constraint
        let valid_max_k = IndexHeader {
            format_version: 3,
            kmer_length: 61,
            window_size: 15,
        };
        assert!(valid_max_k.validate().is_ok()); // k=61 is valid

        let invalid_max_k = IndexHeader {
            format_version: 3,
            kmer_length: 63,
            window_size: 15,
        };
        assert!(invalid_max_k.validate().is_err()); // k=63 exceeds 61

        // Test k+w <= 96 constraint
        let valid_max_kw = IndexHeader {
            format_version: 3,
            kmer_length: 61,
            window_size: 35,
        };
        assert!(valid_max_kw.validate().is_ok()); // k+w=96 is valid

        let invalid_max_kw = IndexHeader {
            format_version: 3,
            kmer_length: 61,
            window_size: 36,
        };
        assert!(invalid_max_kw.validate().is_err()); // k+w=97 exceeds 96

        // Test k must be odd
        let invalid_even = IndexHeader {
            format_version: 3,
            kmer_length: 30,
            window_size: 15,
        };
        assert!(invalid_even.validate().is_err()); // k=30 is even
    }
}

/// Fetch a pre-built index from remote storage
#[cfg(feature = "fetch")]
pub fn fetch(
    index_name: &str,
    kmer_length: u8,
    window_size: u8,
    output: Option<&Path>,
) -> Result<()> {
    const DEFAULT_REPOSITORY_URL: &str =
        "https://objectstorage.uk-london-1.oraclecloud.com/n/lrbvkel2wjot/b/human-genome-bucket/o";

    let base_url = std::env::var("DEACON_REPOSITORY_URL")
        .unwrap_or_else(|_| DEFAULT_REPOSITORY_URL.to_string());

    let filename = format!("{}.k{}w{}.idx", index_name, kmer_length, window_size);
    let url = format!("{}/deacon/{}/{}", base_url, INDEX_FORMAT_VERSION, filename);

    eprintln!("Fetching {}", url);

    let mut response = minreq::get(&url)
        .send_lazy()
        .context("Failed to download index")?;

    if response.status_code != 200 {
        anyhow::bail!("Failed to fetch index: HTTP {}", response.status_code);
    }

    let content_length = response
        .headers
        .get("content-length")
        .and_then(|v| v.parse::<u64>().ok())
        .unwrap_or(0);

    let pb = ProgressBar::new(content_length);

    let output_path = output
        .map(|p| p.to_path_buf())
        .unwrap_or_else(|| PathBuf::from(&filename));

    let mut temp_path = output_path.clone();
    temp_path.as_mut_os_string().push(".tmp");

    let mut file = File::create(&temp_path).context("Failed to create temporary file")?;
    std::io::copy(&mut pb.wrap_read(&mut response), &mut file)
        .context("Failed to write index to file")?;

    std::fs::rename(&temp_path, &output_path).context("Failed to finalise index")?;

    pb.finish_and_clear();
    eprintln!("Index saved to: {}", output_path.display());

    Ok(())
}