On Farm Risk Factors Associated With Goat Milk Quality In Northeast ...

WONDWOSSEN A. GEBREYES

THE OHIO STATE UNIVERSITY

E-MAIL: [email protected]

Capacity Building in Veterinary Public Health and Livestock

Biotechnology: (VPH-BIOTECH GLOBAL)

mailto:[email protected]

Infectious Diseases Molecular Epidemiology Laboratory (IDMEL): http://vet.osu.edu/IDMEL

Foodborne Pathogens Nosocomial pathogens

Salmonella

Campylobacter

Yersinia

Staphylococcus

Acinetobacter

Understanding the Molecular Mechanism

of Antimicrobial Resistance and hyper-virulence “ Farm/ Hospital bed to Genomes ”

Clostridium

http://vet.osu.edu/IDMEL

Goals

Strengthening/building a sustainable capacity for

an integrated food safety, zoonoses, veterinary

public health and Livestock biotechnology using

the “One Health” model.

• Why zoonotic diseases?

•75% of emerging infections

• Why One Health model?

•Prevention of control needs integrated

partnership among disciplines.

Current partners!

VPH Biotech Global

Consortium

Formats for Capacity Building

1. Institutional Training • Modular skill building courses,

2. Technology transfer • Genotyping etc.

3. Graduate fellowship and innovation:

• MSc and PhD levels

4. Knowledge-sharing and awareness • expanding skills and modifying behavior

5. Meta-leadership skill development

• effective interaction among stakeholders

Examples of VPH-Biotech Global activities

Skill building courses:

Molecular Epidemiology of Infectious Diseases

Milk Quality and genotyping projects

Collaborators: OSU, UFPB, UNIVASF, and INSA

Sponsor: OSU Global Gateways Grant

Int’l Congress on pathogens at the Human Animal Interface (ICOPHAI)

MICROBIOLOGICAL QUALITY OF

MILK AND GENOTYPING IN

SEMI-ARID BRAZIL

Genotypic relatedness of staphylococci isolated

from critical control points of small-scale dairy

plants for goat milk pasteurization in semi-arid Brazil

Federal University of Paraiba

College for Agricultural Sciences

Department of Animal Sciences

ICOPHAI 2011

Celso Oliveira 1, Francisca Sousa 1, Melanie Abley 2,

Ariana Meira 1, Wondwossen Gebreyes 2

1 Center for Agricultural Science, Federal University of Paraiba (CCA/UFPB), Brazil

2 College of Veterinary Medicine, The Ohio State University (OSU), USA

Goats and NORTHEAST BRAZIL

95% of goats concentrated in developing

countries

Arid and semi-arid regions

93% of the Brazilian goat herd/ NE Brazil

67% of all goat milk production

Highest rural concentration of impoverished

people in Latin America

67% of population living below the poverty line

Thus limited ability for professional oversight

BACTERIAL CONTAMINANTS OF MILK:

Milk: excellent medium for pathogen growth.

Brucella

Campylobacter

Listeria

Salmonella

Staphylococcus

Tuberculosis

All are zoonotic (infect humans and animals.)

Milk adulteration and “Fome Zero”

E. coli

Shigella

Streptococcus pyogenes

Yersinia enterocolitica

Governmental

Programs Private Market

MILK QUALITY

AND SAFETY

ICOPHAI 2011

Key findings: Less than 40% of the goat milk samples complied with

the national regulations for raw goat milk bacterial threshold.

High prevalence and counts of staphylococci in milk Higher somatic cell count in milk as compared to other

countries. Range from 275,860 cells/ml to 11,055,620 cells/ml

Further investigations: Qualitative impacts of public health: Antimicrobial

resistance Genotypic analysis to determine dissemination patterns.

STAPHYLOCOCCUS SPP.

Staphylococcus spp- 100% prevalence

Range from 500 cfu/ml- 30,900,000 cfu/ml

Mean value= 1,971,206 cfu/ml

ATCC 1 2 3 4 5 6 7 8 9 10 M 11 12 13 14 15 16 17 18 19

20 21 c-

25923

N° CÓDIGO IDENTIFICAÇÃO

ATCC 25923

1 ST 0016/08 Faz. 7

2 ST 0020/08 Faz. 8

3 ST 0040/08 Faz. 14

4 ST 0056/08 Faz. 20

5 ST 0086/08 Faz. 26

6 ST 0087/08 Faz. 26

7 ST 0088/08 Faz. 26

8 ST 0095/08 Faz. 27

9 ST 0096/08 Faz. 27

10 ST 0101/08 Faz. 28

N° CÓDIGO IDENTIFICAÇÃO

11 ST 0102/08 Faz. 28

12 ST 0103/08 Faz. 28

13 ST 0104/08 Faz. 28

14 ST 0110/08 Faz. 29

15 ST 0111/08 Faz. 29

16 ST 0112/08 Faz. 29

17 ST 0124/08 Faz. 31

18 ST 0131/08 Faz. 33

19 ST 0132/08 Faz. 33

20 ST 0140/08 Faz. 35

21 ST 0141/08 Faz. 35

10000

8000

6000

5000

4000

3000

2000

1500

1000

5000

1000

900

800

700

600

500

400

300

200

100

9-16-2010 (82 entries)

Pearson correlation (Opt:1.00%) [0.0%-100.0%]

Rep PCR

10

0

95

90

85

80

75

70

65

60

55

50

45

40

35

30

25

20

15

10

97.7

99.1

78.4

86.8

70.2

93.9

87.9

81.3

67.5

76.5

59.6

63.5

31

77.5

56.9

29.1

27

95.1

82.4

78.5

80.2

57.1

82.1

90.1

66.1

57.4

49.8

79.6

71.4

55.7

37

99

83

92.9

83.5

77

67.2

90.8

98.1

84.5

70

55.1

79.9

96.9

72.9

49

89.7

83

47.8

97.6

94.4

88.7

90.9

85.2

80.7

98.5

97.3

85.7

98

92.5

87.6

80.8

75.6

99.5

98.4

72.3

79.2

56.6

48.8

41.3

33.4

18.9

98.9

90.9

96.4

93.3

82.6

16.4

70.6

41.1

5.1

Rep PCR

Key

P7S9a

P7S9b

CM36a

CM36b

CC2b

CM35

P2S9a

P2S9b

CCC02

PM10a

CC6

NM11

CM29

PS4

CS29

P6S4b

CS23

CM30

PS03a

PS03c

S. aureus c

S. aureus b

PM12

PS03b

P3S10a

P6S4a

NM11a

NM11b

PM11a

CS22

P6S2

PM11b

P3S10b

NM3a

NM3b

CC8

P6S7

S. aureus d

S aureus a

CPC02b

CPC02a

PM10b

P2M3a

P2M3b

CCC02a

CC10

NS9a

CC1

CC2a

NS9b

PS08

NS9c

NS2

P6M3b

P8S2

P6M3a

CCC03a

CCC03b

NM12b

CCM04c

CPM04b

CPM04a

PS9

NM10

NS8a

NS4a

NS4b

NS03a

NS03c

NS03b

NS8b

NS8c

NM12a

P6M11a

P6M11b

P8M3a

CPC03a

CPC03b

P8M3b

CC7

P8S4

P3S8

Dairy Plant

Sumé

Sumé

Monteiro

Monteiro

Cabaceiras

Monteiro

Sumé

Sumé

Cabaceiras

Monteiro

Cabaceiras

Monteiro

Monteiro

Sumé

Sumé

Sumé

Monteiro

Sumé

Sumé

ATCC 2593

ATCC 25923

Monteiro

Sumé

Sumé

Sumé

Monteiro

Monteiro

Monteiro

Sumé

Sumé

Monteiro

Monteiro

cabaceiras

Sumé

ATCC 25923

ATCC 25923

Cabaceiras

Cabaceiras

Monteiro

Monteiro

Monteiro

Cabaceiras

Cabaceiras

Sumé

Cabaceiras

Cabaceiras

Sumé

Sumé

Sumé

Sumé

Monteiro

Sumé

Monteiro

Cabaceiras

Cabaceiras

Monteiro

Monteiro

Monteiro

Monteiro

Sumé

Monteiro

Sumé

Sumé

Sumé

Sumé

Sumé

Sumé

Sumé

Sumé

Monteiro

Monteiro

Monteiro

Monteiro

Cabaceiras

Cabaceiras

Monteiro

Cabaceiras

Sumé

Sumé

Source

Hand swab from dairy plant worker


Raw milk

Raw milk

Raw milk

Raw milk

Equipment of dairy plant

Equipment

Raw milk

Pasteurized milk

Raw milk

Raw milk

Raw milk

Pasteurized milk

Raw milk

Raw milk

Raw milk

Pasteurized

Pasteurized

Pasteurized

Pasteurized



Raw milk

Raw milk

Pasteurized

Raw milk

Hand of worker of dairy plant

Raw milk.

Raw milk.

Raw milk


Pasteurized milk

Pasteurized

Pasteurized milk



Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Pasteurized

Raw milk

Raw milk.




Raw milk

Raw milk.

Raw milk

Raw milk

Pasteurized

Pasteurized

Pasteurized milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk




Pasteurized milk

Pasteurized milk


Raw milk

Pasteurized milk

Equipment

Coagulase

-

-

-

+

-

-

-

-

-

-

-

+

-

-

-

-

+

+

+

-

+

-

+

+

-

-

-

-

-

-

-

+

-

-

-

-

-

-

+

+

+

+

-

-

+

-

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

-

+

-

-

-

-

-

-

-

-

+

9-16-2010 (82 entries)

Pearson correlation (Opt:1.00%) [0.0%-100.0%]

Rep PCR

10

0

95

90

85

80

75

70

65

60

55

50

45

40

35

30

25

20

15

10

97.7

99.1

78.4

86.8

70.2

93.9

87.9

81.3

67.5

76.5

59.6

63.5

31

77.5

56.9

29.1

27

95.1

82.4

78.5

80.2

57.1

82.1

90.1

66.1

57.4

49.8

79.6

71.4

55.7

37

99

83

92.9

83.5

77

67.2

90.8

98.1

84.5

70

55.1

79.9

96.9

72.9

49

89.7

83

47.8

97.6

94.4

88.7

90.9

85.2

80.7

98.5

97.3

85.7

98

92.5

87.6

80.8

75.6

99.5

98.4

72.3

79.2

56.6

48.8

41.3

33.4

18.9

98.9

90.9

96.4

93.3

82.6

16.4

70.6

41.1

5.1

Rep PCR

Key

P7S9a

P7S9b

CM36a

CM36b

CC2b

CM35

P2S9a

P2S9b

CCC02

PM10a

CC6

NM11

CM29

PS4

CS29

P6S4b

CS23

CM30

PS03a

PS03c

S. aureus c

S. aureus b

PM12

PS03b

P3S10a

P6S4a

NM11a

NM11b

PM11a

CS22

P6S2

PM11b

P3S10b

NM3a

NM3b

CC8

P6S7

S. aureus d

S aureus a

CPC02b

CPC02a

PM10b

P2M3a

P2M3b

CCC02a

CC10

NS9a

CC1

CC2a

NS9b

PS08

NS9c

NS2

P6M3b

P8S2

P6M3a

CCC03a

CCC03b

NM12b

CCM04c

CPM04b

CPM04a

PS9

NM10

NS8a

NS4a

NS4b

NS03a

NS03c

NS03b

NS8b

NS8c

NM12a

P6M11a

P6M11b

P8M3a

CPC03a

CPC03b

P8M3b

CC7

P8S4

P3S8

Dairy Plant

Sumé

Sumé

Monteiro

Monteiro

Cabaceiras

Monteiro

Sumé

Sumé

Cabaceiras

Monteiro

Cabaceiras

Monteiro

Monteiro

Sumé

Sumé

Sumé

Monteiro

Sumé

Sumé

ATCC 2593

ATCC 25923

Monteiro

Sumé

Sumé

Sumé

Monteiro

Monteiro

Monteiro

Sumé

Sumé

Monteiro

Monteiro

cabaceiras

Sumé

ATCC 25923

ATCC 25923

Cabaceiras

Cabaceiras

Monteiro

Monteiro

Monteiro

Cabaceiras

Cabaceiras

Sumé

Cabaceiras

Cabaceiras

Sumé

Sumé

Sumé

Sumé

Monteiro

Sumé

Monteiro

Cabaceiras

Cabaceiras

Monteiro

Monteiro

Monteiro

Monteiro

Sumé

Monteiro

Sumé

Sumé

Sumé

Sumé

Sumé

Sumé

Sumé

Sumé

Monteiro

Monteiro

Monteiro

Monteiro

Cabaceiras

Cabaceiras

Monteiro

Cabaceiras

Sumé

Sumé

Source



Raw milk

Raw milk

Raw milk

Raw milk


Equipment

Raw milk

Pasteurized milk

Raw milk

Raw milk

Raw milk

Pasteurized milk

Raw milk

Raw milk

Raw milk

Pasteurized

Pasteurized

Pasteurized

Pasteurized



Raw milk

Raw milk

Pasteurized

Raw milk


Raw milk.

Raw milk.

Raw milk


Pasteurized milk

Pasteurized

Pasteurized milk



Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Pasteurized

Raw milk

Raw milk.




Raw milk

Raw milk.

Raw milk

Raw milk

Pasteurized

Pasteurized

Pasteurized milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk

Raw milk




Pasteurized milk

Pasteurized milk


Raw milk

Pasteurized milk

Equipment

Coagulase

-

-

-

+

-

-

-

-

-

-

-

+

-

-

-

-

+

+

+

-

+

-

+

+

-

-

-

-

-

-

-

+

-

-

-

-

-

-

+

+

+

+

-

-

+

-

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

-

+

-

-

-

-

-

-

-

-

+

PFGE and MLST Genotyping

Unique MLST= ST313 was common

MILK-MILK ADULTERATION

TWO BANDS= Goat milk adulterated with bovine milk.

G= Only goat milk B= Only bovine milk

Test limit of detection= 0.5% (Lane 3).

G B

Dice (Tol 1.0%-1.0%) (H>0.0% S>0.0%) [0.0%-100.0%]

Brazil Staph Aureus100

80

60

40

92.3

50

35.4

100

100

55.8

87.5

75.1

86.7

82.1

77.7

56.7

50.8

36

33.5

Brazil Staph Aureus

Key

ST00104

ST00108

ST00094

ST00075

ST00877

ST00878

ST00293

ST00296

ST00356

ST00357

ST00266

ST00284

ST00313

ST00304

ST00325

ST00375

Origin

USA

USA

USA

USA

USA

USA

Brazil

Brazil

Brazil

Brazil

Brazil

Brazil

Brazil

Brazil

Brazil

Brazil

Host

Feline

Canine

Equine

Equine

Bovine

Bovine

Bovine

Bovine

Bovine

Bovine

Bovine

Bovine

Bovine

Bovine

Bovine

Bovine

Spp.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

R-pattern

AmAnAxChEnErOxMrPbPn

AmAnAxChEnErLnOxMrPbPn

AmAxChGmOxPbPnTbTr

AmAxChGmOxPbPnTr

AmOxPbPn

AmOxPbPn

OxPbPn

AmCfOxPbPn

Ox

Ox

AmAxOxPn

AmAxOxPbPnTe

ChOxPbPnTe

AmOxPbPnTe

AmOxPn

AmOxPbPnTe

mecA

Yes

Yes

Yes

Yes

ND

ND

Yes

Yes

Yes

Yes

Yes

Yes

Yes

Yes

Yes

Yes

blaZ

ND

ND

ND

ND

ND

ND

Yes

Yes

No

No

Yes

Yes

No

Yes

Yes

Yes

PBP2'

ND

ND

ND

ND

ND

ND

Yes

Yes

Yes

Yes

Yes

Yes

Yes

Yes

Yes

Yes

SCCmec

I & II

I & II

IV

IV

ND

ND

I & IV

I & IV

I & II

I & II

None

I & III

I

I & III

I & III

I

Methiciilin Resistance/ MRSA Common in milk in NE Brazil and similar genotypes

with that of milk from the U.S.

Organizing partners

VPH Biotech Global

Consortium

8 key thematic areas

3 keynote and 8 plenary speakers

45 oral presentations

93 poster presentations

275 Participants from 34 nations 356 online registrants:

following via Video streaming

147 participants received scholarship Full, partial or registration fee sponsorship

Closing remarks

THEMATIC AREAS Enteric diseases, food and waterborne pathogens

Wildlife and interface at human diseases

Respiratory diseases and global impact

Drug discovery and antimicrobial resistance

Parasites and environmental impact

Genomics and molecular epidemiology

Immunology and vaccine development

Policy, capacity building and other significant issues

All slides and videos on http://icophai2011.org

http://icophai2011.org

Sponsors

http://icophai.org

http://icophai.org

ICOPHAI 2013

Brazil

Interested to partner/ listserve/ sponsor:

E-mail: [email protected] or [email protected]



Muito Obrigado!

Asante Sana!

Muchas gracias! በበበበበበበበበበበበበ

Merci Beaucoup!

On Farm Risk Factors Associated With Goat Milk Quality In Northeast ...

Documents