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 New Emerging Enteric Pathogens in
Developing Countries:
Aeromonas in Libya as a Model
Khalifa Sifaw Ghenghesh1, Amal Rahouma1,
and Ezzedin Franka2
Departments of Microbiology and Immunlogy1 and
Family and Community Health2,
Faculty of Medicine, Tripoli University,
Tripoli-Libya
Introduction
• Diarrheal diseases are major cause of
morbidity and mortality in children under 5
years old.
• According to the World Health Organization
(WHO) nearly 5 million children die due to
diarrheal disease annually, with most deaths
occur in developing countries.
Enteric agents associated with
children diarrhea
Agents
Classic
New emerging
------------------------------------------------------------------------------------------Bacteria:
Salmonella
Campylobacter
Shigella
Enterotoxigenic E. coli
Enteropathogenic E. coli
Enteroinvasive E. coli

Aeromonas
Enterohemorrhagic E. coli
Enteroadherent E. coli

-----------------------------------------------------------------------------------------Viral:
Rotavirus
Norwalk virus
Adenovirus
Astrovirus
Norovirus
------------------------------------------------------------------------------------------Parasitic:
Entamoeba histolytca
Cryptosporidium
Giardia lamblia
Aeromonas spp.
• Gram-negative, rod-shaped bacteria.
• Associated with gastroenteritis in children less
than five years of age.
• Associated with food-borne and water-borne
outbreaks as well as nosocomial outbreaks.
Aeromonas spp.
• Taxonomy:
– Family Aeromonadeceae.
– At least 14 phenospecies and genospecies
(hybdridization groups [HG]).
– A. hydrophila, A. caviae and A. veronii bv sobria
(A. sobria) predominates in clinical, food and
water samples.
Aeromonas spp.
• Virulence Factors:
– Exotoxins
– Invasiveness
– Adherence
– Others

• Antibiotic Resistance and Treatment:
– Resisatnce to ampicillin

• Laboratory Identification:
– Oxidase +ve
Aeromonas spp in Libya
Prevalence of Aeromonas in Libyan children

------------------------------------------------------------------------------------------------------

Patients

No.
tested

No. (%)
Positive

157
157

24(15)
28(18)

Children (Benghazi, 1996-1997):
- with diarrhea
356
- without diarrhea
100

0(0.0)
0(0.0)

Children (Zliten, 2000-2001)
-with diarrhea

169

9(5.0)

Children (Tripoli, 2008):
- with diarrhea

122

3(2.5)

-------------------------------------------------------------------------------------------------------

Children (Tripoli, 1992-1993):
- with diarrhea
- without diarrhea

-------------------------------------------------------------------------------------------------------
Distribution of Aeromonas species among
patients and controls
No. (%) belonging to the following Aeromonas species:

Subjects

No.

hydrophila veronii

tested

caviae

schubertii

Total

bv sobria

------------------------------------------------------------------------------------------Patients

157

5(3.2)

8(5.1)

13(8.3)

1(0.6)

27(17.2)

Controls

157

9(5.7)

7(4.5)

17(10.8)

2(1.2)

35(22.3)

________________________________________________________
Haemolytic and haemagglutination activities of Aeromonas species
isolated from children with diarrhea (cases) and controls.

______________________________________________________________________

Subjects

No. strains
tested

Number of strains (%) positive for:
Haemolysin Haemagglutination
MS*
MR

------------------------------------------------------------------------------------------------------------------

Cases

27

11(41)

15(56)

7(26)#

Controls

35

11(31)

15(43)

1(3)

______________________________________________________________________
*MS= mannose sensitive; MR= mannose resistant.
#(P<0.01; Fisher's exact test).
Occurrence of Aeromonas in Animals in Libya
_______________________________________________
Animal
No. tested
No. (%)
positive
_______________________________________________
Cows
36
0(0)
Camels
200
1(0.5)
Ostriches
6
1(17)*
Domestic dogs
120
13(11)
Domestic cats
15
1(7%)
Houseflies
150
16(11)
Cockroaches
150
2(1.3)
_______________________________________________
*animal with diarrhea.
Aeromonas in Foods in Libya
_____________________________________________
Food
No. tested
No. (%)
positive
--------------------------------------------------------------------------Fish
48
20 (42)
Chicken carcases
290
218 (75)
Meats:
camel
18
12 (67)
beef
50
19 (38)
Total
68
31 (46)
Juices
147
3 (2.1)
Ice cream
160
30 (19)
_____________________________________________
Aeromonas in Drinking Water in Libya
___________________________________________
Type of
No.
No. (%)
Water
tested
positive
---------------------------------------------------------------------Well
1000
487 (49)
Majin
62
37 (60)
Mineral
216
0 (0.0)
Mosques
50
9 (18)
Faskia
56
41 (73)
Water vendors
50
8 (16)
___________________________________________
Genes coding for virulence factors
in Aeromonas from Libya
No (%) positive for

No of
Aeromonas from:
Strains
act
ast
aer
lip
-----------------------------------------------------------------------------------------Children:
- with diarrhea
12
0
0
10(83)
0
- without diarrhea
12
0
0
10(83)
0
Well water

11

4(36)

6(55)

9(82)

0

Chicken
17
0
3(18) 16(94)
0
---------------------------------------------------------------------------------------------Total
52
4(8)
9(17) 45(87)
0
Antibiotic Sensitivity of Aeromonas
Strains Isolated from Different Sources
____________________________________________________

Antibiotic

% Resistance of Aeromonas from
Children
Water
Chicken
(62)*
(40)
(59)

--------------------------------------------------------------------------------------------------------------

Amoxicillin +
clavulanic acid
Chloramphenicol
Ciprofloxacin
Gentamicin
Kanamycin
Nalidixic acid
Tetracycline
Trimethoprimsulphamethoxazole

97

93

100

6
0.0
0.0
3
0.0
11
23

0.0
0.0
0.0
0.0
0.0
5
0.0

12
2
3
9
37
58
53
Aeromonas in Neighboring Countries
• Studies are lacking from neighboring countries.
New Emerging Enteric Pathogens in
Developing Countries
• The Future:
– Newer agents will continue to emerge.
– The morbidity and mortality associated
with children diarrhea will continue and
may get worst in many developing
countries due to new emerging enteric
pathogens.
• AIDS
New Emerging Enteric Pathogens
in Libya
• New emerging enteric pathogens are not uncommon in
Libya.
• Surveillance studies on such pathogens are needed in
the future to determine their prevalence in different
population groups particularly in the young, the old and
the immunocompromised.
• Also, surveillance studies are needed to determine the
prevalence of the new emerging enteric pathogens in
domestic animals, foods and waters.
• Joint surveillance programs with neighboring countries.
• Such surveillance studies will assist in determining their
mode of transmission, control, prevention and
treatment.

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New Emerging Enteric Pathogens-Aeromonas as a Model

  • 1.  New Emerging Enteric Pathogens in Developing Countries: Aeromonas in Libya as a Model Khalifa Sifaw Ghenghesh1, Amal Rahouma1, and Ezzedin Franka2 Departments of Microbiology and Immunlogy1 and Family and Community Health2, Faculty of Medicine, Tripoli University, Tripoli-Libya
  • 2. Introduction • Diarrheal diseases are major cause of morbidity and mortality in children under 5 years old. • According to the World Health Organization (WHO) nearly 5 million children die due to diarrheal disease annually, with most deaths occur in developing countries.
  • 3. Enteric agents associated with children diarrhea Agents Classic New emerging ------------------------------------------------------------------------------------------Bacteria: Salmonella Campylobacter Shigella Enterotoxigenic E. coli Enteropathogenic E. coli Enteroinvasive E. coli Aeromonas Enterohemorrhagic E. coli Enteroadherent E. coli -----------------------------------------------------------------------------------------Viral: Rotavirus Norwalk virus Adenovirus Astrovirus Norovirus ------------------------------------------------------------------------------------------Parasitic: Entamoeba histolytca Cryptosporidium Giardia lamblia
  • 4. Aeromonas spp. • Gram-negative, rod-shaped bacteria. • Associated with gastroenteritis in children less than five years of age. • Associated with food-borne and water-borne outbreaks as well as nosocomial outbreaks.
  • 5. Aeromonas spp. • Taxonomy: – Family Aeromonadeceae. – At least 14 phenospecies and genospecies (hybdridization groups [HG]). – A. hydrophila, A. caviae and A. veronii bv sobria (A. sobria) predominates in clinical, food and water samples.
  • 6. Aeromonas spp. • Virulence Factors: – Exotoxins – Invasiveness – Adherence – Others • Antibiotic Resistance and Treatment: – Resisatnce to ampicillin • Laboratory Identification: – Oxidase +ve
  • 8. Prevalence of Aeromonas in Libyan children ------------------------------------------------------------------------------------------------------ Patients No. tested No. (%) Positive 157 157 24(15) 28(18) Children (Benghazi, 1996-1997): - with diarrhea 356 - without diarrhea 100 0(0.0) 0(0.0) Children (Zliten, 2000-2001) -with diarrhea 169 9(5.0) Children (Tripoli, 2008): - with diarrhea 122 3(2.5) ------------------------------------------------------------------------------------------------------- Children (Tripoli, 1992-1993): - with diarrhea - without diarrhea -------------------------------------------------------------------------------------------------------
  • 9. Distribution of Aeromonas species among patients and controls No. (%) belonging to the following Aeromonas species: Subjects No. hydrophila veronii tested caviae schubertii Total bv sobria ------------------------------------------------------------------------------------------Patients 157 5(3.2) 8(5.1) 13(8.3) 1(0.6) 27(17.2) Controls 157 9(5.7) 7(4.5) 17(10.8) 2(1.2) 35(22.3) ________________________________________________________
  • 10. Haemolytic and haemagglutination activities of Aeromonas species isolated from children with diarrhea (cases) and controls. ______________________________________________________________________ Subjects No. strains tested Number of strains (%) positive for: Haemolysin Haemagglutination MS* MR ------------------------------------------------------------------------------------------------------------------ Cases 27 11(41) 15(56) 7(26)# Controls 35 11(31) 15(43) 1(3) ______________________________________________________________________ *MS= mannose sensitive; MR= mannose resistant. #(P<0.01; Fisher's exact test).
  • 11. Occurrence of Aeromonas in Animals in Libya _______________________________________________ Animal No. tested No. (%) positive _______________________________________________ Cows 36 0(0) Camels 200 1(0.5) Ostriches 6 1(17)* Domestic dogs 120 13(11) Domestic cats 15 1(7%) Houseflies 150 16(11) Cockroaches 150 2(1.3) _______________________________________________ *animal with diarrhea.
  • 12. Aeromonas in Foods in Libya _____________________________________________ Food No. tested No. (%) positive --------------------------------------------------------------------------Fish 48 20 (42) Chicken carcases 290 218 (75) Meats: camel 18 12 (67) beef 50 19 (38) Total 68 31 (46) Juices 147 3 (2.1) Ice cream 160 30 (19) _____________________________________________
  • 13. Aeromonas in Drinking Water in Libya ___________________________________________ Type of No. No. (%) Water tested positive ---------------------------------------------------------------------Well 1000 487 (49) Majin 62 37 (60) Mineral 216 0 (0.0) Mosques 50 9 (18) Faskia 56 41 (73) Water vendors 50 8 (16) ___________________________________________
  • 14. Genes coding for virulence factors in Aeromonas from Libya No (%) positive for No of Aeromonas from: Strains act ast aer lip -----------------------------------------------------------------------------------------Children: - with diarrhea 12 0 0 10(83) 0 - without diarrhea 12 0 0 10(83) 0 Well water 11 4(36) 6(55) 9(82) 0 Chicken 17 0 3(18) 16(94) 0 ---------------------------------------------------------------------------------------------Total 52 4(8) 9(17) 45(87) 0
  • 15. Antibiotic Sensitivity of Aeromonas Strains Isolated from Different Sources ____________________________________________________ Antibiotic % Resistance of Aeromonas from Children Water Chicken (62)* (40) (59) -------------------------------------------------------------------------------------------------------------- Amoxicillin + clavulanic acid Chloramphenicol Ciprofloxacin Gentamicin Kanamycin Nalidixic acid Tetracycline Trimethoprimsulphamethoxazole 97 93 100 6 0.0 0.0 3 0.0 11 23 0.0 0.0 0.0 0.0 0.0 5 0.0 12 2 3 9 37 58 53
  • 16. Aeromonas in Neighboring Countries • Studies are lacking from neighboring countries.
  • 17. New Emerging Enteric Pathogens in Developing Countries • The Future: – Newer agents will continue to emerge. – The morbidity and mortality associated with children diarrhea will continue and may get worst in many developing countries due to new emerging enteric pathogens. • AIDS
  • 18. New Emerging Enteric Pathogens in Libya • New emerging enteric pathogens are not uncommon in Libya. • Surveillance studies on such pathogens are needed in the future to determine their prevalence in different population groups particularly in the young, the old and the immunocompromised. • Also, surveillance studies are needed to determine the prevalence of the new emerging enteric pathogens in domestic animals, foods and waters. • Joint surveillance programs with neighboring countries. • Such surveillance studies will assist in determining their mode of transmission, control, prevention and treatment.