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Portuguese Forensic Institute Tests

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With thanks to Albym

Processo Vol II page 475


Letter from PJ to Institute of National Legal Medicine regarding evidence collected

Fax to the Director of the Police Science Laboratory (LPC) from DIC Portimao dated 8 May 2007
Subject Examinations

The above Inquiry is the investigation into the disappearance of the child Madeleine Beth McCann.
1) This serves to confirm the inspection, already done, of the apartment at the following address:
- Praia da Luz - Ocean Club - Block A - Apartment 5A
2) We ask that the trace evidence recovered at that location be delivered to the INML, namely all those items that may be treated [examined] by thatInstitute.
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Processo Vol II page 479

Letter from PJ to Institute of National legal Medicine regarding evidence collected at Murano Quinta dos Figos

Fax to the Director of the Police Science Laboratory (LPC) from DIC Portimao dated 8 May 2007
Subject Examinations

The above Inquiry is the investigation into the disappearance of the child Madeleine Beth McCann.
1) This serves to confirm the inspection, already done, of the apartment at the following address:
- Praia da Luz - Rua 1st of May/wall of Quinta dos Figos
2) We ask that the trace evidence recovered at that location be preserved for future possible examinations.
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Processos Vol IX

Page 2416-2417



Letter to the National Forensics Institute (INML) from Goncalo Amaral


17th August 2007

The present inquiry is in relation to the investigation of the disappearance of Madeleine McCann on 3rd May from P da L.

The English authorities in possession of elements collected from the family home in Leicestershire, which are being examined in an unidentifiedlaboratory, traced the girl’s DNA profile, in annex to this document.

As it is necessary to the investigation we request the following:

1. Whether in the tests done by the INML Madeleine’s profile was established?

2. In the case of a positive answer to point 1, that it should be determined whether the girl is the daughter of the parents identified – Gerald Patrick McCann and Kate Healy.

3. Information about whether the profile obtained by the INML corresponds to the profile traced in the English laboratory, on the request of the British authorities.

4. If there are differences between the English and Portuguese profiles that a report should be drawn up for this inquiry.

5. In the follow-up of point 4, we request to be informed whether in the English profile the girl is the daughter of the McCann couple.

6. Other information that might be of use to the investigation.

Signed

G. Amaral
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With many thanks to Albym
See also http://www.gerrymccannsblogs.co.uk/PJ/PORTUGUESE-FORENSIC.htm#p9p2307

Vol. 8, pages 2023 to 2027

FIRST INML REPORT 9 JULY 2007


NATIONAL INSTITUTE OF LEGAL MEDICINE
GENETIC AND BIOLOGICAL FORENSIC SERVICES


Processo n? 2007/000268/P-B
Processo n? 2007/000244/CR-B
Processo n? 2007/000226/LX-BC


EXPERT REPORT

IDENTIFICATION OF BIOLOGICAL TRACES AND COMPARATIVE STUDY WITH REFERENCE SAMPLES

INFORMATION


REQUESTING AGENCY: Departamento de Investigacao Criminal de Portimao da Pol?ia Judiciaria e Directoria Nacional.

PROCESSO: NUIPC 201/07.0GALGS
NOME: MADELEINE BETH MCCANN.

OFICIO N.? 013.079 and fax n? 399, both dated 08/05/2007, that say "to proceed to deliver the traces recovered from apartment n? 5-A of the tourist resort "The Ocean Club" in Praia da Luz in Lagos", which were accompanied by 7 (seven) sealed envelopes under a delivery note n? 156/2007-CR/L, as part of the investigation into the disappearance of the child MADELEINE BETH MCCANN.

OFICIO N.? 13443 and fax n? 413 both dated 15/05/2007, that say to proceed to deliver the traces recovered from the residence at Rua do Ramalhete, Casa Liliana in Praia da Luz in Lagos (1 envelope), and from the Volkswagen with numberplate 44-77-KD (7 sealed envelopes), which were accompanied by delivery notes n? 170A/2007CR/L and 170B/2007-CR/L. respectively.

FAX N? 000415 dated 15/05/2007, that requests the INML to proceed with all necessary examinations on all the traces delivered to them as part of the abovementioned Inquiry.

OFICIO N.? 13453 /I /2007 dated 15/05/2007, that says to proceed to deliver the bucal swabs recovered from Matthew David Oldfield, Jane Michelle Tanner, Gerald Patrick Mccann, Kate Marie Healy, with delivery note n? 170C/2007-CR/L and relevant official documents from the Departamento de InvestigacaoCriminal de Portimao da Policia Judiciaria.

OFICIO N.? 001803 dated 17/05/07, that says "to remit bucal swabs recovered from individuals, asking for comparisons of DNA profiles with those from biologicaltraces recovered from various locations, as part of the present legal documents", accompanied by relevant official documents from the Departamento de lnvestigacaoCriminal de Portimao da Pol?ia Judiciaria.

OFICIO N.? 002026 dated 05/06/07, that says "to deliver two bucal swabs provided by Robert James Queriol Eveleigh Murat", with relevant official documents from the Departamento de InvestigacaoCriminal de Portimao da Pol?ia Judiciaria.


MATERIAL RECEIVED


There were received 258 traces and 12 reference samples - bucal swabs from 11 individuals.


DESCRIPTION OF MATERIAL RECEIVED


TRACES
Relating to Apartment n? 5-A in the tourist resort "The Ocean Club" Praia da Luz-Lagos:

- 32 hairs in envelope n? 1 recovered from the floor at the entrance to the children's bedroom.

- 28 hairs in envelope n? 2 recovered from the floor next to the bed from which the child disappeared.

- 4 hair in envelope n? 3 recovered from the top of the bed from which the child disappeared.

- 15 hairs in envelope n? 4 recovered from the floor next to the bed that was next to the window in the children's bedroom.

- 1 piece of cloth in envelope n? 5 recovered from the bedspread of the bed next to the window in the children's bedroom.
Fragment of cloth, mauve/violet in colour with square motifs, circular in form about 10cm in diameter. A small fluorescent spot is observed under a Crime-light.

- 31 hairs in envelope N? 6 recovered from the floor of the lounge.

- 58 hairs in envelope N? 7 recovered from the entrance hall at the front door of the apartment.

[The above were] Delivered by the Pol?ia Judiciaria on 08/05/2007.

Relating to the Volkswagen 44-77-KD:

- 15 hairs in envelope N? 1 recovered from the front-right seat.

- 2 hairs in envelope N? 2 recovered from the front-left seat.

- 12 hairs in N? 3 recovered from the floor in front of the front seats.

- 7 hairs in envelope N? 4 recovered from the floor in front of the rear seat.

- 4 hairs in envelope N? 5 recovered from the rear seat.

- 5 hairs in envelope N? 6 recovered from the rear part of the vehicle.

- 4 hairs in envelope N? 7 recovered from the internal covering of the roof.

Relating to Residencia Casa Liliana - Praia da Luz-Lagos:

- 40 hairs in envelope N? 1 recovered from the bed in the bedroom of of the suspect.

[The above were] Delivered by the Policia Judiciaria on 15/05/2007.

REFERENCE SAMPLES

- 4 sealed bucal swabs from:
Matthew David Oldfield, Jane Michelle Tanner, Gerald Patrick McCann, Kate Marie Healey.

Delivered by the Pol?ia Judiciaria on 15/05/2007.

- 6 sealed bucal swabs from:
Dianne Webster, Rachael Mariamma Jean Mampilly, Russell James O'Brien, Fiona Payne, David Anthony Payne, Silvia Maria Correia Batista.

Delivered by the Pol�cia Judiciaria on 18/05/2007.

- 2 sealed bucal swabs from:
Robert James Queriol Eveleigh Murat.

Remitted by the Departamento de Investigacao Criminal de Portimao da Pol?ia Judiciaria and delivered on 06/06/2007.

METHODOLOGY

1-Morphology analysis of hairs

- Macro- and microscopic characterisation.

2-Nature of the sample

Acid Phosphatase Test to detect semen on the small spot on the cloth fragment in envelope n? 5 recovered from the bedspread of the bed next to the window of the children's bedroom.
[Ref: http://www.semenonpanties.com/laux.htm ]

3- DNA extraction:

The following methods were used to extract DNA from the material received for analysis:

- Extraction with phenol-chloroform (organic extraction) [1,2]
[Ref: http://en.wikipedia.org/wiki/Phenol-chl ... extraction ]
- Tissue and Hair Extraction kit -"DNA IQ(tm) System" (Promega)
- "QIAamp micro DNA kit" method (Qiagen)
- Chelex method [3]

[1] Huhne, J., Pfiffer, H., Waterkamp, K., Brinkmann, B. 1999. "Mitochondrial DNA in human hair shafts existence of intra-individual differences'". International Journal of Legal Medicine (1999), 112: 172-175.
[2] Maniatis T. Fritsch EF, Sambrook J. 1982. "Molecular Cloning - A Laboratory Manual". Cold Spring Harbor Laboratory.
[3] Walsh P.S., Metzer D.A., Higuchi (1991). Chelex-100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 10: 506-513.

4-Nuclear DNA study

After extraction the following genetic markers were studied by PCR:

? Autosomic STRs: D8S1179, D21S11, D7S820, CSFlPO, D3S1358, HUMTH01, D13S317, D16S539, D2S1338, D19S433, HUMVWA31/A, TPOX, D18S51, D5S818, HUMFIBRA/FGA and Amelogene, using the primers and conditions of amplification suggested in the kit AmpFISTR Identifiler (Applied Bioystems).

? Autosomic STRs: D3S1358, HUMTH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSFlPO, Penta D, HUMVWA31/A, D8S1179, TPOX, HUMFIBRA/FGA, and Amelogene using the primers and the conditions ofamplification suggested in kit PowerPlex(tm)16 System (Promega).

5-Mitochondrial DNA study

The following methodology was used in the Mitochondrial DNA study:

PCR Amplification:

PCR Amplification of the human mtDNA regions HVI and HVII, using specified primers and amplification conditions in accordance with Vigilant et al., 1989 [4] e Wilson et al., 1995 [5], Alvarez-lglesias et al., [6].

[4] Vigilant L, Pennington R. Harpending 11, Kocher TD, Wilson AC. 1989. "Mitochondrial DNA Sequences in Single Hairs from a Southern African Population". Proc Natl Acad Sci, USA, 1989, Vol. 86: 9350 - 9354.
[5] Wilson MR, DiZinno JA, Polanskey D, Replogle J. Budowle B. 1995. "Validation of Mitochondrial DNA Sequencing for Forensic Casework Analysis". Int J Legal Med. 1995,108: 68 - 74.
[6] Alvarez-Iglesias V, Jaime JC, Carracedo A, Salas A.2007. "Coding region mitochondrial DNA SNPs:Targeting East Asian and Native American haplogroups" Forensic Sci. Int. Genetics, 2007, 1 :44-55.

Sequenciation:
Direct sequenciation of the HVI e HVll segments by capilar electrophoresis in "ABI PRISM 3100 ou 3130 Genetic Analyser" (Applied Biosystems) automated sequenciator with fluorescent detection using the following kits:

- "BigDye(R) Terminator vl.1 Cycle Sequencing" (Applied Biosystems)
- "BigDye(R) Terminator v1.3 Cycle Sequencing" (Applied Biosystems)
- "dRhodamine Terminator Cycle Sequencing Ready Reaction" (Applied Biosystems)

The programme "SeqScape(R) Software Version 2.0" (Applied Biosystems) was used for comparison of the sequences obtained from the analysed samples with the CRS (Cambridge Reference Sequence).


RESULTS


1- Morphology analysis of the hairs

All hairs received for analysis were subjected to macro- and microscopic characterisation, with the majority of those having the root being in a telogenic phase.
[Ref: https://www.promega.com/geneticidproc/e ... c/ab20.pdf ]

Relating to Apartment n? 5-A

- Envelope N? 1 - "recovered from the floor at the entrance to the children's bedroom" 25 hairs with root; 4 hairs only stem; 3 hairs non-human.

- Envelope N? 2 - "recovered from the floor next to the bed from which the child disappeared" 3 hairs with root; 25 hairs only stem.

- Envelope N? 3 - "recovered from the top of the bed from which the child disappeared" 2 hairs with root; 2 hairs only stem.

- Envelope N? 4 - " recovered from the floor next to the bed that was next to the window in the children's bedroom" ll hairs with root; 4 hairs only stem.

- Envelope n? 6 - "recovered from the floor of the lounge" 26 hairs with root; 4 hairs only stem; l hair non-human.

- Envelope n? 7 - "recovered from the entrance hall at the front door of the apartment" 44 hairs with root; 8 hairs only stem; 6 hairs non-human.

Relating to the Volkswagen 44-77-KD

- Envelope n? 1 - "recovered from the front-right seat" 13 hairs with root; 1 hair root only; 1 hair non-human.

- Envelope n? 2 - "recovered from the front-left seat" 2 hairs with root.

- Envelope n? 3 - "recovered from the floor in front of the front seats" 9 hairs with root; 2 hairs only stem; 1 hair non-human.

- Envelope N? 4 - "recovered from the floor in front of the rear seat" 1 hair with root; 6 hairs only stem.

- Envelope N? 5 - "recovered from the rear seat" 4 hairs only stem.

- Envelope N? 6 - "recovered from the rear part of the vehicle" 1 hair with root; 4 hairs only stem.

- Envelope N? 7 - "recovered from the internal covering of the roof" 3 hairs with root; 1 hair only stem.

Relating to the Residencia Casa Liliana - Praia da Luz-Lagos:

- Envelope N? 1 - "recovered from the bed in the bedroom of of the suspect" 36 hairs with root; 4 hairs only stem.

2-Nature of the sample

Acid Phosphatase Test to detect semen on the small spot on the cloth fragment in envelope n? 5 recovered from the bedspread of the bed next to the window of the children's bedroom: Weakly positive.





3 - Nuclear DNA study

Autosomic STRs:

- were obtained from: hair root CEnv 7-51 recovered in the apartment (entrance hall-Env.7), hair root PVT07 recovered in the vehicle (floor in front of rear seat Env.4), hair root C742-05 recovered in the vehicle (floor in front of the front seat- Env. 3) and the small spot on the piece of cloth (recovered from the bedspread of the bed next to the window in the children's bedroom Env.5).

- were obtained from swabs:

Matthew David Oldfield, Jane Michelle Tanner, Gerald Patrick McCann, Kate Marie Healey, Dianne Webster, Rachael Mariamma Jean Mampilly, Russell James O'Brien, Fiona Payne, David Anthony Payne, Silvia Maria Correia Batista e Robert James Queriol Eveleigh Murat.

4 - Mitochondrial DNA study

- Mitochondrial DNA was obtained from 194 vestiges - 1 spot and 193 hairs (roots and/or stems) of the 246 analysed. It was not possible to obtain results from 52 hairs, 31 of which were recovered in the apartment.


Mitochondrial DNA profiles of the vestiges




Input check

Sample Identification Perfil S
Database Release 1
Maximum differences displayed 5
Include partially overlapping profiles no
Consider(multiple)insertions in the
polyC-stretches at 16193, 309 and 573
individual differences as:
Selected ranges 73-340
16024-16365

mtDNA query profile 73A>G
263A>G
315.1C
16256C>T
16270C>T

Source Forensic data

Selected populations Africa: ALL
East Asia: ALL
South East Asia: ALL
West Eurasla: ALL

OUTPUT
Selected ranges: 73-340
16024-16365

mtDNA query profile: 73A>G, 263A>G, 315.1C, 16256C>T, 16270C>T

OVERALL RESULT SUMMARY (4527/4527)

DIFFERENCES TO NUMBER OF CUMULATIVE
QUERY PROFILE HAPLOTYPES NUMBER OF HAPLOTYPES
0 8 8
1 40 48
2 54 102
3 178 280
4 442 722
5 595 1317
>5 3210 4527

Africa RESULT SUMMARY (348/348)

East Asia RE5ULT SUMMARY (162/162)

South East Asia RESULT 5UMMARY (187/187)

West Eurasia RESULT SUMMARY (3830/3830)

DIFFERENCES TO NUMBER OF CUMULATIVE
QUERY PROFILE HAPLOTYPES NUMBER OF HAPLOTYPES
0 8 8
1 39 47
2 54 101
3 178 279
4 429 708
5 584 1292
>5 2538 3830

http://www.empop.org 19-05-2008

As I understand it, the EMPOP database is the global repository of regional profiles of mitochondrial DNA (mtDNA).
The person doing the query entered the specific mt DNA characteristics of JT's hair into the query form and found 8 exact matches in the database, i.e. at least eight maternal lines having the identical Haplotype to JT exist in Europe.
There were no exact matches in the other three regional groups.

Because of this result the inquiry could not say with absolute certainty that the hair found in Burgau actually came from JT because it could have come from any person born of a mother belonging to any of the eight profiles in the database - one profile, of course, had to be of JT's maternal line, so it could have been some distant (or close) relative.
CODE
mtDNA Number of Comparison with
profile Samples Recovered from reference samples

B 3 3 entrance hall
B1 1 1 entrance hall
-------------------------------------------------------
13 lounge
15 entrance hall
C 53 13 entrance to bedroom Haplotype of Kate Healy (Mother)
4 floor; bed next to window
1 bed of the child
7 floor; next to child's bed
-------------------------------------------------------
D 1 1 lounge
D1 1 1 entrance to bedroom
D1 1 1 floor; bed next to window
-------------------------------------------------------
F 3 1 lounge
2 entrance hall
-------------------------------------------------------
G 1 1 lounge Haplotipo de Matthew Oldfield
I 1 1 entrance hall Haplotipo de David Payne
-------------------------------------------------------
1 floor; bed next to window
J 3 1 entrance hall
1 floor; next to child's bed
-------------------------------------------------------
K 14 1 Residencia Liliana
13 vehicle(envlps 1,3,4,5 and 6)
-------------------------------------------------------
1 spot on bedspread
L 8 1 entrance to bedroom
2 entrance hall
4 floor; bed next to window
-------------------------------------------------------
M 48 13 vehicle (envlps 1,2,3 and 4)
35 Residencia Liliana Haplotipo de Robert Murat
-------------------------------------------------------
2 lounge
11 entrance hall
N 24 6 entrance to bedroom Haplotipo de Gerald Mccann
2 floor; bed next to window
3 floor; next to child's bed
-------------------------------------------------------
0 2 1 lounge
1 entrance hall Haplotipo de Russell O'Brien
-------------------------------------------------------
Q 2 1 entrance hall
1 lounge
-------------------------------------------------------
Profiles different from each other and from those above:
28 17 of which were found in the apartment.
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Post  Info Mon 8 Mar - 11:17

With many thanks to Albym
See also http://www.gerrymccannsblogs.co.uk/PJ/P ... tm#p9p2307

Vol XII Pages 3252 to 3253

From Goncalo Amaral

To: INML

Date: 4th September 2007

The present request deals with the investigation into the disappearance of Madeleine McCann on 3rd May 2007.

Within the scope of this request we have asked the INML to carry out various analyses as your letter 2007000226 LX ? BC reports.


The results were delivered to us in a letter dated 9th July 2007 ref 13.079, with a cover letter dated 11th July 2007.

Upon reading the report we observed that several DNA profiles are defined but which have not been identified.


The apartment from which the samples were collected for analysis is occupied by families from the UK.

The UK police and legal authorities have informed us about the existence of a database for the register of DNA profiles.

As the objective is to proceed to identify the profiles defined in your analyses we request that you send us the list of unidentified profiles.
This request has the objective of seeking their eventual identification through the UK legal authorities.

Goncalo Amaral
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Vol. 16 pages 4358 to 4361
Information re Haplotype-S with rest results

Extract from a letter from INML to the Inquiry clarifying the Haplotype S conundrum:
61
===============================
Em resposta ao oficio de V. Exmo. acima referenciado, cumpre-nos informar que um perfil de ADN mitocondrial nao e exclusivo e objectivamente identificativo de uma determinada pessoa. Salienta-se que apos consulta da base de dados de sequencias da regiao controlo do ADN mitocondrial, EMPOP database, os resultados obtidos permitiram a identificacao de 8 haplotipos identicos ao haplotipo S (sem diferences), num universo de 3830 amostras do grupo populacional 'West Eurasia", que compreende as populacoes da Europa.

Junto se anexa a consulta efectuada para melhor esclarecimento:
================================
[Rough translation]
In response to your letter I advise that a mitochondrial DNA profile is not an exclusive or objective identification of a single [determined] person. Consulting the regional control sequences in the EMPOP database we find 8 results that are identical (zero differences) to the Haplotype S sample, from a total population [universe] of 3830 samples in the 'West Eurasia' population group, known as the populations of Europe.

See the printed consultation below:
============================

* Bl and Dl have a different sequence from B and D.


CONCLUSIONS


1st- The macro- and microscopic analysis of 257 hairs revealed 245 human and 12 non-human.

2nd- With respect to autosomic STRs the genetic profiles of the following analysed samples matched with the reference sample indicated::

- hair root CEnv 7-51 recovered in the apartment (entrance hall-Env.7) matched with Gerald McCann, father of the victim.

- hair root PVT07 recovered in the vehicle (floor in front of rear seat Env.4) match with Robert James Queriol Eveleigh Murat.

3rd- With respect to autosomic STRs the female genetic profile identified from hair root C742-05 recovered in the vehicle (floor in front of the front seat- Env 3) did not match any profile from the reference samples.

4th- With respect to autosomic STRs the male genetic profile identified from the spot on the cloth fragment (bedspread of the bed next to window in children's bedroom- Env 5) did not match any profile from the reference samples.

5th- In the samples from apartment 5-A, several mitochondrial DNA profiles were found:

- Profile identified by letter "C", present in 53 samples, was identical to that of Kate Healy, mother of the victim, meaning those samples were from her or from someone having the same maternal bloodline.

- Profile identified by letter "N", present in 24 samples, was identical to that of Gerald McCann, father of the victim, meaning those samples were from him or from someone having the same maternal bloodline.

- Profile identified by letter "G", present in 1 samples, was identical to that of Matthew David Oldfield, meaning that sample was from him or from someone having the same maternal bloodline.

- Profile identified by letter "I", present in 1 samples, was identical to that of David Anthony Payne, meaning that sample was from him or from someone having the same maternal bloodline.

- Profile identified by letter "O", present in 2 samples, was identical to that of Russell James O'Brien, meaning those samples were from him or from someone having the same maternal bloodline.

- Profile identified by letter "L", present in the spot on the bedspread of the bed next to the window and in seven hairs, meaning that all these samples came from the same person or from someone having the same maternal bloodline, did not match any of the reference samples.

- Profiles identified by letters "B", "D", "F", "J" and "Q" are different from the above, and from each other, and are distinct from reference samples.

6th- In the samples analysed from Residencia Liliana and the vehicle, several mitochondrial DNA profiles were found:

- Profile identified by letter "M", present in 48 samples (35 from Residencia Liliana and 13 from the vehicle), was identical to that of Robert James Queriol Eveleigh Murat meaning those samples were from him or from someone having the same maternal bloodline.

- Profile identified by letter "K", present in 14 samples (1 from Residencia Liliana and 13 from the vehicle), meaning that all these samples came from the same person or from someone having the same maternal bloodline, did not match any of the reference samples.

- One of the 13 samples from the vehicle in letter "K" - hair root C742-05 (floor in front of the front seat - Env 3) showed a female DNA genetic profile.

7th- The remaining 28 samples analysed, of which 17 were recovered from the apartment, showed mitochondrial DNA from different from each other, and distinct from those above.

8th- Of the 245 hairs analysed, no results were obtained from 52.

National Institute of Legal Medicine, 9 July 2007.

[Titles, names and signatures of signatories to the report]
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With many thanks to Albym.
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OUTROS APENSOS I - volume II pages 284-299

Comparative analysis report of biological vestiges collected from various locales 2008.03.31

GENETIC & BIOLOGICAL FORENSIC SERVICES


Processo n* 2007/000565/PT-B
Processo n* 2007/000244/CR-B5,B6
Processo n* 2007/000226/LX-BC1



EXPERT REPORT

IDENTIFICATION OF BIOLOGICAL VESTIGES [remains] AND COMPARATIVE STUDY WITH REFERENCE SAMPLES

INFORMATION


REQUESTING ENTITY: Department of Criminal Investigation of Portimao of the Pollcia Judiciaria and National Directorate.

PROCESSO: NUIPC 201/07.0GALGS
NAME: MADELEINE BETH MCCANN.

OFFICIAL LETTER N.' 003610, dated 31/10/2007, that solicited the performance of lawful examinations of various samples of hair recovered from the following locations:

- Apartment C, 2' Floor, of Aparthotel Sol e Mar, in Burgau
- Vehicle type Audi, model A4, licence-plate 10-91-PF
- Vehicle type Renault, model Kangoo, licence-plate 07-20-UI
- Vehicle type Volkswagen, model Passat, licence-plate 57-12-HP
- Sofas located in in the garage of the CID Portimao
- Garden of apartment 5A, block A, tourist resort "OCEAN CLUB", Praia da Luz, Lagos.

OFFICIAL LETTER N.' 004080, dated 12/12/2007, that solicited "examinations as appropriate, with the view to determine the nature of the mark (found on the counterpane [quilt/bedspread] of one of the beds in apartment 5A of the tourist resort OCEAN CLUB in Praia da Luz) and if it permits the identification of the profile indicated by letter (L)".

OFFICIAL LETTER N.' 004079, dated 12/12/2007, that says "to send an item marked as fibre and a plastic item both extracted from the car boot..., material already examined..., soliciting the indulgence of the reader in the sense that further examinations are necessary on the aforementioned material, using a method considered adequate for the detection of possible biological remains that may not have been detected in the first examination to which they were subjected."

1

GENETIC & BIOLOGICAL FORENSIC SERVICES

Processo n* 2007/000565/PT-B
Processo n* 2007/000244/CR-B5,B6
Processo n* 2007/000226/LX-BC1

OFFICIAL LETTER N.* 000550, dated de 04/07/07, that says to send 12 oral swabs [swatches] and hair collections, accompanied by respective letters of authorisation for their collection duly prepared and signed in the Departamento de Investigacao Criminal de Portimao da Policia Judiciaria in respect of the following people: Silvia Maria Correia Ramos Baptista (head-hair only), Mario Domingos Moreira, Luis Filipe Monteiro Ferro, Dina Maria dos Reis Rocha, Marina Paula Ramos Batista Castela, Maria Julia Serafim da Silva, Jose Maria Batista Roque, Jose Leal Pimentel, Antonio Conceicao Duarte, Nelson Filipe Pacheco da Costa, Maria Irene Trovao Ferro, Joao Francisco Pascoa Luis Trigo Barreiras e Vitor Manuel Martins.

OFFICIAL LETTER N.* 00551, dated 04/07/07, that says to send two oral swabs [swatches] and two capillary collections, obtaine from Michaela Walczuch e de Luis Filipe Duarte Antonio, accompanied by statements of swab collection and capillary collection duly prepared and signed in the Departamento de Investigacao Criminal de Portimao da Policia Judiciaria.

MATERIAL RECEIVED

In this 2nd fase, 187 hairs [generic], 2 pieces from the car boot and reference samples from 15 individuals were received.

In the 1st fase, were received 258 vestigios and 12 reference samples - oral swabs [swatches] from 11 individuals. The vestigios included 257 hairs [generic] and one speck detected in a cloth fragment collected from the counterpane of the bed next to the window of the bedroom of the children in apartament 5A (envelope 5), as mentioned in the previous report.

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DESCRIPTION OF THE MATERIAL RECEIVED

VESTIGIOS delivered by Policia Judiciaria on 08/11/2007:

With respect to Aparthotel Sol e Mar, 2', apartment C - Burgau

- 8 labelled envelopes with delivery note n' 420A/2007-CR/L:
Vestigio:
Vg1B- "Sundry head-hair collected from the kitchen floor of the apartment" - 20 hairs.
Vg2 - "Sundry head-hair collected at the entrance of the apartment" - 23 hairs.
Vg3 - "Sundry head-hair collected from the bedroom floor of the apartment" - 27 hairs.
Vg4 - "Sundry head-hair collected from the bed in the bedroom of the apartment" - 17 hairs.
Vg5 - "Sundry head-hair collected from the bathroom of apartment" - 16 hairs.
Vg6 - "Sundry head-hair collected from the floor of lounge of apartment" - 43 hairs.
Vg7 - "Sundry head-hair collected from a sofa presently in the lounge of the apartment"- 3 hairs.
Vg8 - "Sundry head-hair collected from a sofa presently in the lounge of the apartment"- 3 hairs.

With respect to Vehicle Audi A4 10-91-PF:

- 1 labelled envelope with delivery note n' 420C/2007-CR/L:
Vestigio:
Vg1 - "Sundry head-hair collected from the front seat, right side"- 3 hairs.

With respect to Vehicle Renault Kangoo 07-50-UI:

- 3 labelled envelopes with delivery note n' 420E/2007-CR/L:
Vestigio:
Vg1 - "Sundry head-hair collected from the row of seats at the rear of the Vehicle"- 1 hair.
Vg2 - "Sundry head-hair collected from the small area at the rear of Vehicle" - 2 hairs.
Vg3 - "Sundry head-hair collected from the front seat, right side" - 1 hair.

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With respect to Vehicle Volkswagen Passat 57-12-HP:

- 1 labelled envelope with delivery note n' 420F/2007-CR/L:
Vestigio:
Vg1 - "Sundry head-hair collected from the rear seat" - 6 hairs.

with respect to the sofas that are in the garage of the DIC Portimao:

- 3 labelled envelopes with delivery note n' 420B/2007-CR/L:
Vestigio:
Vg1 - "Sundry head-hair collected from the sofa having one seat"- 7 hairs.
Vg2 - "Sundry head-hair collected from the sofa having two seats"- 2 hairs.
Vg3 - "Sundry head-hair collected from the sofa having one seat" - 2 hairs.

With respect to the garden area of apartment 5A, tourist resort "OCEAN CLUB", Praia da Luz, Lagos:

- 1 labelled envelope with delivery note n' 420D/2007-CR/L:
Vestigio:
Vg25 - "Sundry head-hair" - 11 hairs.

VESTIGIOS delivered by Pol?ia Judiciaria on 14/12/2007:

With respect to the car boot:

- Pieces that included a fibre and plastic, having been previously analysed in various areas.

REFERENCE SAMPLES delivered by Pol?ia Judiciaria on 04/07/2007:

- 14 oral swabs [swatches] and collections of head-hair, relating to:

Mario Domingos Moreira, Luis Filipe Monteiro Ferro, Dina Maria dos Reis Rocha, Marina Paula Ramos Batista Castela, Maria Julia Serafim da Silva, Jose Maria Batista Roque, Jose Leal Pimentel, Antonio Conceicao Duarte, Nelson Filipe Pacheco da Costa, Maria Irene Trovao Ferro, Joao Francisco Pascoa Luis Trigo Barreiras, Vltor Manuel Martins, Michaela Walczuch e Luis Filipe Duarte Antonio.

- Collection of head-hair relatiing to:

Silvia Maria Correia Ramos Baptista.

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METHODOLOGY

1- Morphologic Analysis of the hairs

- Macro- and microscopic characterisation.

2- Nature of the sample

Phadebas Forensic test, for detection of saliva on the fragment of cloth corresponding to vestigio n'. 5 collected from the counterpane of the bed next to the window of the children's bedroom.

3- DNA Extraction:

DNA was extracted from the material received for analysis using one of the foloowing methods:

- Tissue and Hair Extraction kit-"DNA IQ(TM) System" (Promega)
- "QIAamp micro DNA kit" (Qiagen)
- Metodo de Chelex [1]

[1] Walsh P.S., Metzer D.A., Higuchi (1991). Chelex-100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 10: 506-513.

4- Study of the nuclear DNA

After extraction the following genetic markers were studied using PCR:

- Autosomic STRs: D8S1179, D21Sl l, D7S820, CSFIPO, D3S1358, HUMTH01, D13S317, D16S539, D2S1338, D19S433, HUMVWA31/A, TPOX, D18S51, D5S818, HUMFIBRA/FGA and Amelogene, using primers and amplification conditions recommended in kit AmpFISTR(TM) Identifiler (Applied Bioystems).

- Autosomic STRs: D3S1358, HUMTH01, D21SII, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSFIPO, Penta D, HUMVWA31/A, D8S1179, TPOX, HUMFIBRA/FGA, and Amelogene using primers and amplification conditions recommended in kit PowerPlex(TM) 16 System (Promega).

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5 - Estudo do DNA mitocondrial

The mitochondrial DNA study (HVRI e HVRII) was performed in accordance with the following metodology:

PCR Amplification:
PCR amplification of the HVI e HVII regions of the human mtDNA, using specified primers amplification conditions in accordance with Vigilant et al., 1989 [2], Wilson et al., 1995 [3] e Alvarez-lglesias et al., [4].

[2] Vigilant L, Pennington R. Harpending H. Kocher TD, Wilson AC. ]989. "Mitochondrial DNA Sequences in Single Hairs from a Southern African Population". Proc Natl Acad Sci, USA, 1989, Vol. 86: 9350 - 9354.
[3] Wilson MR, DiZinno JA, Polanskey D, Replogle J. Budowle B. 1995. "Validation of Mitochondrial DNA Sequencing for Forensic Casework Analysis". Int J Legal Med. 1995, 108: 68 - 74.
[4] Alvarez-lglesias V, Jaime JC, Carracedo A, Salas A.2007. "Coding region mitochondrial DNA SNPs:Targeting East Asian and Native American haplogroups" Forensic Sci. Int. Genetics, 2007, 1 :44-55.

Sequenciation: Direct sequenciation of the HVI e HVII segments by capillary electrophoresis in the automatic sequenciator "ABI PRISM 3100 ou 3130 Genetic Analyser" (Applied Biosystems) with detection by fluorescence, using one of the kits:

- "BigDye(R) Terminator vl.1 Cycle Sequencing" (Applied Biosystems)
- "BigDye(R) Terminator v1.3 Cycle Sequencing" (Applied Biosystems)
- "dRhodamine Terminator Cycle Sequencing Ready Reaction" (Applied Biosystems)

The program "SeqScape(R) Software Version 2.0" (Applied Biosystems) was used to compare the sequences obtained from the analysed samples with the CRS (Cambridge Reference Sequence).

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RESULTS

[b]1- Morphologic analysis of the hairs

All the hairs received for analysis were subjected to macro- and microscopic characterisation. It was observed that some hairs were only stems while other had both root and stem. The majority of those with roots were found to be telogenic.

2- Nature of the sample

Phadebas Forensic test, for detection of saliva on the fragment of cloth corresponding to vestigio n'. 5 collected from the counterpane of the bed next to the window of the children's bedroom: Positive.

3- Study of the nuclear DNA

Autosomic STRs results were obtained from the following vestigios: root of one of the hairs collected from the bedroom floor of the Burgau apartment (Vg3) and in the lower zone of te fibre from the car boot. Table 1 includes, also, the profile of the spot from the counterpane, already sent previously.

Table 1 - Autosomic STRs of vestigios.


Code:
From the bedroom Vestigios inferior Spot on the cloth
SYSTEMS floor- Apart. in zone of the fibre fragment from the
Burgau -Vg 3 from the car boot counterpane
Amelogenina XY XY XY
D8S1179 13-15 10-11 12-13
D21S11 28-30 29 29-32.2
D7S820 9-10 10-12 10-12
CSFIPO 10-11 10-13 10-11
D3S1358 15-18 14-16 15-18
HUMTH01 6-9.3 9-9.3 9.3
D13S317 11 11-12 12-14
D16S539 11-12 10-14 11
D2S1338 17 18-24 18-25
D19S433 15 13-14 13-15
HUMVWA31/A 18-19 17-18 18
TPOX 8-11 8 11
D18S51 16-18 13-14 17-18
D5S818 12 11-13 12-13
HUMFIBRA/FGA 20 21-25 20-24
Penta E 7-8
Penta I> 9

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Translation:

GENETIC AND BIOLOGICAL FORENSIC SERVICES

Processo n' 2007/0005651PT-B
Processo n' 2007/000244/CR-B5,B6
Processo n' 2007/000226/LX-BCI

Autosomic STRs were also obtained relating to reference samples from:

Mario Domingos Moreira, Luis Filipe Monteiro Ferro, Dina Maria dos Reis Rocha, Marina Paula Ramos Batista Castela, Maria Julia Serafim da Silva, Jose Maria Batista Roque, Jose Leal Pimentel, Antonio Conceicao Duarte, Nelson Filipe Pacheco da Costa, Maria Irene Trovao Ferro, Joao Francisco Pascoa Luis Trigo Barreiras, Vitor Manuel Martins, Michaela Walczuch, Luis Filipe Duarte Antonio e Silvia Maria Correia Ramos Baptista.

4- Study of mitochondrial DNA

The vestigios delivered on 08/11/2007 and on 14/12/2007:

Mitochondrial DNA results were obtained in 137 hairs (roots and/or stems) from the 187 analysed. It was not possible to obtain any results from 46 hairs, and in 4 cases incomplete sequences were obtained.

5- Total of the samples studied:

Since May 2007, the following samples were analysed in the GENETIC AND BIOLOGICAL FORENSIC SERVICES offices of the North, Central e South Delegations of the Instituto Nacional of Medicina Legal:

- 25 reference samples.
- 447 vestigios, comprising:
>> 444 hairs, of which 432 were human;
>> 1 mark on a counterpane;
>> 2 objects from a car boot.

Of the 432 human hairs (roots and/or stems), 315 provided the results reflected in Table 2.

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Tabela 2. Haplotypes of Mitochondrial DNA of the vestigios and comparison with reference samples
Code:
Haplotypes No. of Collection point of vestigios Comparison with
samples reference samples

B 4 3 Entrance hall Ocean(Vg7),
1 Entrance hall Burgau(Vg2)

B1 1 1 Entrance hall Ocean(Vg7) Haplotype of Irene Ferro (IF)

13 Lounge floor Ocean(Vg6)
14 Entrance hall Ocean(Vg7)
C 42 3 Bedroom entrance Ocean(Vg1) Haplotype of Kate Healy (KH) and
1 Child's bed Ocean(Vg3) Haplotype of Jose Roque (JMBR)
4 Floor next to bed next window Ocean(Vg4)
7 Floor next to child's bed Ocean(Vg2)
C* 11 1 Entrance hall Ocean(Vg7)
10 Bedroom entrance Ocean(Vg1)

D 1 1 Lounge floor Ocean(Vg6)

D1 2 1 Bedroom entrance Ocean(Vg1)
1 Floor next to bed next window Ocean(Vg4)

E 1 1 Entrance hall Ocean(Vg7)
Ea 1 1 Sofa 1-seat; CID garage(Vg1)

F 3 1 Lounge floor Ocean(Vg6) and Haplotype of Vitor Martins (VM)
2 Entrance hall Ocean(Vg7) Haplotype of Vitor Martins (VM)

G 1 1 Lounge floor Ocean(Vg6) Haplotype of Matthew Oldfield (MO)

I 1 1 Entrance hall Ocean(Vg7) Haplotype of David Payne (DP)

1 Floor next to bed next window Ocean(Vg4)
J 3 1 Entrance hall Ocean(Vg7)
1 Floor next to child's bed Ocean(Vg2)
J1 0 Haplotype of Ant6nio Duarte(ACD)

K 16 1 Residencia Liliana(Vg1)
13 vehicle Volkswagen Envl(3), Env3(5), Haplotype of Michaela Walczuch (MW)
Env4(2), Env5(1), Env6(2) and of Luis Ferro (LFMF)
1 Renault(Vg3) e 1 Passat(Vg1)


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Tabela 2. Haplotypes of DNA Mitocondrial dos vestigios e comparacao com as samples of referencia enviadas (cont.)
Code:
Haplotypes No. of Collection point of vestigios Comparison with
samples reference samples

L 4 1 Mark counterpane bed (saliva)(env5)
3 Floor next to bed next window Ocean(Vg4)
La 4 1 Bedroom entrance Ocean(Vgl)
2 Entrance hall Ocean(Vg7)
1 Floor next to bed next window Ocean(Vg4)
L2 1 1 Vehicle Volkswagen (Env5)

M 27 13 Vehicle Volkswagen Envl(8), Env2(1), Env3(2), Env4(2)
13 Residencia Liliana(Vgl)
1 Bathroom Burgau(Vg5) Haplotype of Robert Murat (RQMU)
M* 22 22 Residencia Liliana(Vgl)

2 Lounge floor Ocean(Vg6)
10 Entrance hall Ocean(Vg7)
N 20 5 Bedroom entrance Ocean(Vgl) Haplotype of Gerald McCann (GM)
1 Floor next to bed next window Ocean(Vg4)
2 Floor next to child's bed Ocean(Vg2)
N* 4 1 Bedroom entrance Ocean(Vgl), 2 junto janela Ocean(Vg4)
1 Corridor Ocean(Vg7)

1 Lounge floor Ocean(Vg6),1 entrance hall
O 2 Ocean(Vg7) Haplotype of Russell O'Brien(RJB)

P 2 2 Garden Ocean(Vg5)

Q 2 1 Entrance hall Ocean(Vg7), 1 lounge floor Ocean(Vg6)

R 0 Haplotype of Nelson Costa (NFPC)
R* 1 1 Vehicle Volkswagen Env4

S 2 1 Bathroom (Vg5) Burgau and 1 Sofa (Vg7) Burgau Haplotype of Jane Tanner (JT)

T 8 1 LoungeFloor(Vg6), 2 Kitchen(Vgl),
2 EntHall(Vg2) Burgau, 3 BedFloor(Vg3) Burgau

Ta 2 1 Fl Kit(Vgl) and 1 Bedroom Floor(Vg3) Burgau

U 2 1 Entrance hall(Vg2) Burgau and 1 bed(Vg4) Burgau


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Tabela 2. Haplotypes of DNA Mitocondrial dos vestigios e comparacao com as samples of referencia enviadas (cont.)
Code:
Haplotypes No. of Collection point of vestigios Comparison with
samples reference samples

V 1 1 Lounge floor Ocean(Vg6)
V* 1 1 Renault(Vg2)

X 2 1 Bedroom entrance Ocean(Vgl) and 1 sofa 1-seat; garage(Vgl)

Y 2 1 sofa 1-seat; garage(Vgl) and 1 corridor Ocean(Vg7)

Z 3 l Residencia Liliana(Vgl)
2 sofa 1-seat garagem(Vgl e Vg3)

17 Ent.Hall(Vg2), 17 Kit.Floor(Vgl), 20 BedroomFloor(Vg3)
AA 85 Brg
13 LoungeFl(Vg6), 4 bathroom(Vg5),10 Bed&bedroom(Vg4)
Burgau
4 Sofa(Vg7 e 8) Burgau

AAa 6 3 Bed&bedroom(Vg4) and 3 bathroom(Vg5) Burgau

AA* 9 7 LoungeFloor(Vg6) Burgau
2 Vehicle Passat(Vgl)

AB 2 2 Audi A4(Vgl)


15 Haplotypes different from each other and from those above:
Code:
1 Bedroom entrance Ocean(Vgl) .
1 Bed, main bedroom Burgau(Vg4)
1 Sofa of 1-seat; garage(Vgl)
1 Floor next to bed next to window Ocean(Vg4)
1 Sofa of 2-seats; garage(Vg2)
1 Entrance hall Burgau(Vg2)
1 Bathroom Burgau(Vg5)
1 Sofa of 2-seats; garage(Vg2)
1 Bedroom floor Burgau(Vg3)
1 Entrance hall Ocean(Vg7)
1 Lounge floor Ocean(Vg6)
1 Garden Ocean(Vg25)
1 Vehicle Volkswagen (Env5)
1 Bathroom Burgau(Vg5)
1 Entrance hall Ocean(Vg7)
a Difference in 309.1C with respect to the corresponding Haplotype * Results obtained solely in HVI
1 One difference in regio HVII with repsect to the corresponsing Haplotype
2 Two differences in regio HVII with repsect to the corresponsing Haplotype

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CONCLUSIONS

First: In total in the two phases of the case there were 25 reference samples and 447 vestiges studied: 1 spot on the counterpane; 2 objects from the car boot and 444 hairs. The macro- and microscopic analyses of the 444 hairs revealed that 432 were humano and 12 non-human, no result being obtained for 98 hairs and only partial results for l9 hairs. With respect to the plastic object from the car boot no results were obtained.

Second: The search for saliva in the mark on the counterpane from one of the beds in apartament 5A of tourist resort Ocean Club was positive.

Third: The study of autosomic STRs from the sample received in the 2 phases revealed:

There were matches between the reference sample genetic profiles and the following analysed samples:

- hair root (CEnv 7-51) collected in apartament Ocean Club (entrance hall env.7) and Gerald Mccann, father of the victim.

- hair root (PVT07) collected in the Volkswagen 44-77-KD (floor in front of rear seats env.4) and Robert James Queriol Eveleight Murat.

- hair root (C742-05) collected in Volkswagen 44-77-KD (floor in front of front seats - env.3) and Michaela Walczuch.

There were no matches between the reference sample genetic profiles and the following analysed samples:

- Mark on the cloth fragment collected from apartament Ocean Club (counterpane of the bed next to the window of the children's bedroom - env. 5).

- Hair root collected in the apartament in Burgau (bedroom floor - vg3).

- Vestigios in the lower zone of the fibre from the car boot.

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Fourth: The mitochondrial DNA study revealed (a):

In the analysed samples from apartament 5-A of the Ocean Club:

- The Haplotype identified by the letter B1, present in one sample, and identical to that of Maria Irene Trovao Ferro (IF), meaning that sample was from her or an individual of the same maternal bloodline.

- The Haplotype identified by the letters C e C*, present in 53 samples, and identical to that of Kate Healy (KH), mother of the victim and to that of Jose Maria Batista Roque (JMBR), meaning those sample were from those persons or individuals of the same maternal bloodlines.

- The Haplotype identified by the letter F, present in 3 samples, and identical to that of Vltor Manuel Martins (VM), meaning those samples were from that person or individuals of the same maternal bloodlines.

- The Haplotype identified by the letter G, present in one sample, and identical to that of Matthew David Oldfield (MO), meaning that sample was from that person or an individual of the same maternal bloodline.

- The Haplotype identified by the letter I, present in one sample, and identical to that of David Anthony Payne (DP), meaning that sample was from that person or an individual of the same maternal bloodline.

- The Haplotype identified by the letters N e N*, present in 24 samples, and identical to that of Gerald Mccann (GM), father of the victim, meaning those sample were from that person or individuals of the same maternal bloodlines.

- The Haplotype identified by the letter O. present in 2 samples, and identical to that of Russell James O'Brien (RJB), meaning those samples were from that person or individuals of the same maternal bloodlines.

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In the analysed samples from other locations:

- The Haplotype identified by the letter K, present in 16 samples, (1 in the Residencia Liliana, 13 in vehicle Volkswagen, 1 in the Renault and l in the Passat) and identical to that of Michaela Walczuch (MW) and to that of Luis Filipe Monteiro Ferro (LFMF), meaning those samples were from those persons or individuals of the same maternal bloodlines.

- The Haplotype identified by the letters M e M*, present in 49 samples, (35 in the Residencia Liliana, 13 in the vehicle Volkswagen and 1 in the bathroom of the apartament in Burgau), and identical to that of Robert James Queriol Eveleight Murat (RQMU), meaning those samples were from that person or individuals of the same maternal bloodline.

- The Haplotype identified by the letter R*, present in one sample from the vehicle Volkswagen, and identical in HVI, to that of Nelson Filipe Pacheco da Costa (NFPC) - letter R, meaning that sample was from that person or an individual of the same amternal bloodline.

- The Haplotype identified by the letter S, present in 2 samples, (apartment in Burgau), and identical to that of Jane Michelle Tanner (JT), meaning those samples were from that person or individuals of the same maternal bloodline.

5a- The study of the mitochondrial DNA revealed also (a):

- The Haplotypes identified by letters D, D1, J, L, La, P e Q are different from those above, having been found solely in samples from apartament 5-A Ocean Club, not corresponding to any of the reference sample Haplotypes.

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- The Haplotypes identified by the letters L2, T, Ta, U e AB are also different from those above, having been found in the apartament in Burgau or in one of the vehicles, not corresponding to any of the reference sample Haplotypes:

Haplotype L2 - Vehicle Volkswagen
Haplotypes T, Ta - apartament in Burgau
Haplotype U - apartament in Burgau
Haplotype AB - Vehicle Audi A4

- The Haplotypes identified by the letters B, E, Ea, V, V*, X, Y, Z, AA, AAa e AA* are different from those above and distinct from the reference samples, having the same Haplotype been found in distinct locations:

Haplotype B - apart. Ocean Club and apart. Burgau
Haplotypes E, Ea - apart. Ocean Club and sofa of 1-seat in the garage in Portimao
Haplotypes V, V* - apart. Ocean Club and vehicle Renault
Haplotype X - apart. Ocean Club e sofa of 1-seat in the garage in Portimao
Haplotype Y - apart. Ocean Club e sofa of 1-seat in the garage in Portimao
Haplotype Z - Residencia Liliana e sofa of 1-seat in the garage in Portimao
Haplotypes AA, AAa, AA* - apartment in Burgau and vehicle Passat

- The Haplotype identified by the letter J1 corresponds to Antonio Conceicao Duarte (ACD).
This Haplotype was not identified in any of the analysed vestigios.
____________________________
(a) The Haplotypes C*, M*, N*, R*, V* e AA* are identical in the region HVI to the corresponding Haplotype, there not having been obtained any results in region HVIT.
The Haplotypes Ea. La, Ta e AAa presented a difference in 309.1C in relation to that of the corresponding Haplotype.
The Haplotypes Bl, Dl, Jl e Ll presented a difference in the region HVII in relation to that of the corresponding Haplotype.
The Haplotype L2 presented two differences in the region HVII in relation to that of the corresponding Haplotype.

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- The remaining 15 analysed vestigios reflected 15 Haplotypes different from each other and distinct from all of the above(including the reference samples).

Instituto Nacional of Medicina Legal, 31 of Marco of 2008

[centre]Director da Delegacao do Centro e Vogal do Conselho Directivo

Prof. Doutor Francisco Corte Real

Directora of Service - Delegacao do Norte

Prof. Doutora Maria of Fatima Pinheiro

Directora of Service - Delegacao do Centro

Dra. Maria Joao Anjos Porto

Directora of Service - Delegacao do Sul

Dra. Rosa Maria Espinheira
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(Note: these examinations refer to the material collected at the Burgau apartment)


Processos Vol XIII
Pages 3476-3478


Forensic Examination Report

Nº 200707143 – BG

Requested by: Portimao DIC

Process Number: 201/07 OGALGS
731 of 28th September 2007
618 and 619 of 2nd November 2007
034308 of 4th October 2007
Email message with annex sent on 8th November 2007

Delivery Guides: 189/2007- CRL of 24th May 2007
196/2007 of 29th May 2007

Examination initiated on: 8th November 2007.


Material for examination

1. One swab referenced as “Vestige nº1 from the floor next to the wall.
2. One swab referenced as Vestige nº 2 from the top of the wall.
3. One swab referenced as Vestige nº 3 from the floor next to the wall.
4. One swab references as Vestige nº 1 A from the door of the kitchen cupboard.

Requests

The examination of the haematic material received for examination is requested with respect to the DNA analysis and a comparative study with the profiles sent via email on 8th November and in letter nº 034308 of 4th October 2007

Observations and Tests Carried Out

The swabs described in lines 1, 2 and 3 were stained with brownish red.

The swab described in line 4 was very slightly stained in a yellow tone.

Preliminary Tests

Search for peroxidase existing in the blood using the Kastle Mayer test.

DNA Analysis

Methods used for DNA extraction.

Extraction of DNA from the blood samples using the organic method (Phenol/Chloroform).
- 1 – 4 swabs

Amplification of DNA using the P.C.R. technique.

The amplification of the DNA was carried out using the P.C.R technique (Polymerase Chain Reaction) and was analysed according to the following:

- D3S1358, VWA, D16S539, D2S1338, Amelogenina (sex), D8S1179, D21S11, D18S51,
- D19S433, TH01 and FGA using primers marked with fluorochromes and amplification conditions in accordance with the protocol described in AmpFISTR SGM Plus PCR amplification Kit.

Detection and Analysis of Amplified DNA

The detection and analysis of the amplified DNA was carried out in the automatic sequencer ABI Prism Genetic Analyser.

Conclusion

- In the swabs described in lines 1, 2 and 3, blood vestiges were found.
- In the swab described in line 4 very slight traces of blood were found.
- In the DNA analysis of the haematic vestiges detected in the swabs described in lines 1 and 2 a profile was obtained belonging to an individual of the male sex.
- In the DNA analysis of the swab described in line 3 only some results were obtained from the loci that were studied. According to the results obtained a mixture of biological traces belonging to more than one individual were found, it only being possible to indicate the profile of the main contributor who presented an identity of polymorphisms for a set of loci studied with the profile described in the previous line.
- There is no identity of polymorphisms of the haematic vestiges detected in the swabs described in lines 1, 2 and 3 with the profiles sent by email on 8th November 2007 and in the letter nº 034308 of the 4th October 2007.
- In the DNA analysis of the haematic vestiges detected in the swab described in line 4 results were only obtained from some of the loci studied, corresponding to the profile of an individual of the female sex. According to the results obtained, there is no polymorphism identity with the profiles sent by email on 8th November 2007 and in the letter nº 034308 of the 4th October 2007.

Return of material: return the remains of the material received for examination.

15th November 2007


Signed

Technician Checked by
Superior Specialist Superior Specialist
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Post  Info Mon 8 Mar - 11:18

Processos Vol XIII
Page 3591—Translated cover letter re: Eeyore pyjamas for testing (no date)

http://www.gerrymccannsblogs.co.uk/P13/ ... e_3591.jpg
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Post  Info Mon 8 Mar - 11:19

From Processos Vol XVII

Pages 4515 – 4516


Names taken from PJ list of activities that are in the files.

Forensic Section

Buccal swabs taken:


Gerald McCann Page 904
Matthew Oldfield Page 908
Jane Tanner Page 932
Russell O’Brien Page 942
Rachel Mampilly Page 948
Dianne Webster Page 956
Silvia Baptista Page 1291
Fiona Payne Page 1326
David Payne Page 1327
Robert Murat Page 1658
Michaela Walczuch Page 1714
Luis Antonio Page 1716
Kate Healy Page 1804
Maria Ferro Page 1828
Joao Barreiras Page 1829
Vitor Martins Page 1830
GNR Officers Page 1831 –1834
Maria Morreira Page 2298
Luis Ferro Page 2299
Dina Rocha Page 2300
Marina Castela Page 2301
Maria da Silva Page 2302
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Processos Vol VII
Page 1713


To: Police Scientific Laboratory
Lisbon

5th June 2007

Subject: Sending of Pyjamas

The present inquiry investigates the disappearance of Madeleine McCann on 3rd May 2007. I am herewith delivering to the Police Scientific Laboratory a pair of girl’s pyjamas.

The Pyjamas are from Marks and Spencers, size 2 – 3 years -97 cm.

The pyjamas are composed of two pieces: camisole type without buttons and half sleeves, pink with designs, letters and tracing in white with (small) floral patterns, the right pyjama bottom leg has a design (smaller size) which is identical to that of the camisole.

The pyjamas being sent are “equal” in make, model, size, colours and designs as well as presumably the texture, to those the little girl was wearing at the time of her disappearance. The article sent serves for eventual comparisons with fibres collected by the competent officers of the Police Scientific Lab, within the scope of the current investigation.

With compliments

Signed

The Coordinator of the Criminal Investigation

Goncalo Amaral
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