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DISTRIBUTION OF PSEUDOMONAS AERUGINOSA AND AEROMONAS HYDROPHILA IN
GROUNDWATER IN ALGERIA
HAFFARESSAS Y., AYAD N., BOUSSAYOUD R., MOUFFOKF. Pasteur Institute of Algeria, Dely-Brahim, Alger e-mail: [email protected]
Abstract
A microbiological study was conducted for one year and nine months on groundwater in Algeria. 175 groundwater samples were taken during the period of January 2017 / September 2018 at the level of 31 Cities in Algeria. In this study the Membrane Filtration technique was used and the microbiological quality of the groundwater shows a high frequency of pathogenic bacteria. Abundances of Pseudomonas aeruginosa and Aeromonas hydrophila vary and undergo wide spatio-temporal fluctuations. The very high number of microbial germs in water is a significant health risk for an Algerian population who drink water regularly.
Keywords: Underground water, P.aeruginosa, A.hydrophila, Microbiological quality.
Introduction. Groundwater is a vital resource for humanity, whose vulnerability to microbial pollution is known. In these waters, there is a relatively diversified fauna and microflora [3, 20], the stygobal microflora being largely bacterial [14]. The immediate environment of these organisms has variable effects on them. Many microbial communities live in the soil and in aquatic environments; their physiology in these two types of biotopes are often similar, although their nutrient sources are significantly different [25]. Various factors are likely to affect the development of microorganisms in an environment. In the aquatic environment, the physicochemical and biological properties significantly affect the survival of bacteria, the man who plays the role of "matter" and "water of environment" [6]. As "middle water", biotope water and allows the development of microorganisms; the water plays the role of the solvent in time, along with that of the reagent concerning the elements and chemical compounds present. These elements and chemical compounds in most cases, condition the survival of microorganisms present. Bacteria are usually the most abundant microorganisms in nature. Their unwanted presence in a medium indicates a greater or lesser health risk to humans, the pathogenicity of usually harmless bacteria may occur as a result of immunodepression of the host. Many bacteria described as opportunistic pathogens belong to the genera Pseudomonas, Aeromonas, Staphylococcus, Vibrio, Legionella, Plesiomonas, Klebsiella, Serratia and Acinetobacter [12, 15, 20]. Pseudomonas aeruginosa and Aeromonas
hydrophila found in soil, in aquatic and hospital environments, are responsible for diarrhea, wound infections and genitourinary and ocular infections in immunocompromised individuals [15]. Some bacteria, for example Aeromonas salmonicida and Streptococcus uberis, pose a health risk for many aquatic animals [1, 4, 18]. The nature and degree of influence of environmental factors on the distribution and survival of micro-organisms are generally very varied. Environmental temperature, CO2 and dissolved organic matter content and the nature and concentration of nutrients available may affect growth rates of microorganisms present. These parameters are, among other things, factors of variability in the productivity of bacterial communities [17, 20, 26, 30]. Iodides and quaternary ammonium compounds can inhibit the development of Listeria monocytogenes, L. ivanovii and L. innocua [20, 27] under certain environmental conditions. Changes in dissolved oxygen content can significantly affect the physiology of bacteria metabolizing nitrites and ammonia nitrogen [10, 20]. This work aims to identify the 2 bacteria: P. aeruginosa and A. hydrophila; two opportunistic pathogenic bacteria, in different underground water eco-systems located in Algeria.
The genus Aeromonas. Aeromonas are gram negative bacilli, positive oxidase, and facultative anaerobes. They are environmental germs, ubiquitous, living preferentially in water and soil and responsible for sometimes very serious infections. The genus Aeromonas is notable for its antibiotic resistance profile, its wide range of virulence factors, the complexity and confusion
surrounding its taxonomy, and the great phenotypic variability between and within species. Today, it is composed of 28 species, six of which are redundant or controversial and of which only 13 have been isolated in the clinic. Of these, the most frequently clinically isolated species are, in order of decreasing frequency, Aeromonas veronii biovar sobria, Aeromonas hydrophila, Aeromonas caviae, Aeromonas aquariorum, Aeromonas jandaei, Aeromonas schubertii. Types of infection, extremely diverse, include wound infections, bacteraemia, gastroenteritis, peritonitis, hepatobiliary, respiratory, or ophthalmic infections, which occur rather during the summer months. These are rare but emergent infections occurring in immunocompromised patients (cancerous or hepatobiliary pathology) or in immunocompetent patients. Aeromonas may be associated with nosocomial infections, particularly through the use of leeches in reconstructive surgery. Identification at the species level is based, in order of increasing performance, on biochemical methods, mass spectrometry, identification by sequencing of household genes. The natural resistance to betalactamines is characterized by the production of one to three enzymes of chromosomal origin, of cephalosporinase, oxacillinase or
metallocarbapenemase type, inducible and of coordinated expression. Aeromonas are usually sensitive to fluoroquinolones, aminoglycosides (excluding tobramycin), tetracyclines and tigecycline. They are inconsistently sensitive to tobramycin and cotrimoxazole. Treatment is based on a third-generation cephalosporin or fluoroquinolone with or without gentamicin in a severe setting and co-trimoxazole for diarrhea requiring antibiotic treatment [13].
Natural Resistance. Aeromonas generally produces three co-inducible chromosomal beta-lactamases: Class C cephalosporinase (AsbA1 in A. sobria, AmpC homolog of P. aeruginosa). Class D oxacillinase (AsbB1 in A. sobria, inhibited by clavulanic acid and also called OXA-12). Class B imipenemase: CphA in A. hydrophila AsbM1 in A. sobria (also called ImiS) ^ All strains of Aeromonas are naturally resistant to ampicillin (but naturally sensitive to imipenem despite the presence of imipenemase) [23, 24, 28, 29]. Example: A. hydrophila (fig. 1).
AcquiredResistance. Extended-spectrum Class A beta-lactamases (ESBL): The beta-lactamase TEM-12 was identified in a strain of Aeromonas caviae on a self-transferable plasmid. Curiously, the enzyme is not expressed in A. caviae and its detection was
only possible after transfer of the plasmid into an E. coli recipient strain [23, 28].
;05 ж Ж CF
Ж СП Ш ЮХ
m Alt ICC PI?
CK CAZ АШ CP
Figure 1. Resistance to aixipicillxu and imipeneiiiase
Pseudomonas aeruginosa. Pseudomonas aeruginosa occupies a central position in the current problem of nosocomial infections. This ubiquitous gram-negative bacillus is responsible for 10% to 15% of all nosocomial infections, a higher frequency being reported in certain categories of high-risk patients: chronic bronchopulmonary diseases (especially cystic fibrosis), immunodepression (neutropenia). acquired immunodeficiency syndrome), severely burned, patients hospitalized in intensive care [21]. P. aeruginosa is particularly involved in 20 to 30% of pneumonia acquired through mechanical ventilation. Although the severity of P. aeruginosa nosocomial infections is determined by the species-specific virulence and co-morbidities of the patients concerned, it also depends on the pathogen's ability to accumulate antibiotic resistance mechanisms and the therapeutic difficulties that arise result. These acquired resistances are in addition to the many natural resistances of the species and may concern all active classes on wild strains. Colistin is currently the main option for the treatment of multidrug-resistant P. aeruginosa infections [5]. This cyclic bactericidal polypeptide remains in effect active on almost all of these strains [7].
Natural Resistance. Pseudomonas is naturally resistant to a large number of antibiotics due to the production of a class C inducible chromosomal beta-lactamase [31] which is not inhibited by clavulanate and which preferentially hydrolyzes first-generation cephalosporins, and poor membrane permeability. For the beta-lactam family, the molecules that remain active are ticarcillin, piperacillin, cefsulodine, the combination ticarcillin + clavulanate, ceftazidime, aztreonam and imipenem (example: P. aeruginosa 1746). P. aeruginosa is also generally resistant to kanamycin.
Acquired Resistance. Beta-lactam resistance in Pseudomonas is often a serious problem because it
often leads to resistance to most antibiotics. This type of resistance is generally linked to mutations leading to hyperexpression of class C chromosomal beta-lactamase [31] (for example, P. aeruginosa 2307) and to a decrease in membrane permeability (defeat of porin D2 specifically associated with resistance to imipenem) (eg, P.aeruginosa 1092). Numerous beta-lactamases (TEM, OXA, PSE), conferring resistance to penicillins, have been identified in P. aeruginosa.
Beta-lactamases of the ESBL type. PER-1 beta-lactamase, which confers an ESBL-like resistance phenotype, has been described in a clinical strain of P. aeruginosa. More recently, TEM (TEM-42) and SHV-2 variants have also been characterized. Numerous oxacillinase mutants [8] (OXA-11, -13, -14, -15, -18, for example), conferring a high level of resistance to third-generation cephalosporins and inhibited by clavulanate, have been described in recent years.
Resistancetoimipenem. (exampleP.aeruginosa1092). It can be non-enzymatic. It then results from the loss of porin D2 which results in a specific decrease in permeability for imipenem. Resistance is expressed at a high level only if the loss of D2 is accompanied by a hyperproduction of the chromosomal cephalosporinase.
It can be enzymatic. It is in this case related to the production of class B metallo-beta-lactamases [9] (zinc enzymes) which generally have a broad substrate spectrum including third-generation cephalosporins (except aztreonam) and imipenem. These enzymes are inhibited by EDTA. At present, they are found in a plasmid situation, which makes them fear their dissemination in the near future.
Resistance by modification of PLPs. Modifications of PLP3 have been described in strains resistant to cefsulodin. An imipenem-resistant strain showed changes in PLP4 [2].
Resistance by efflux mechanism. In P. aeruginosa, an efflux system (MexAB-OprM) has been described. An increase in the expression level of these proteins may lead to a decrease in beta-lactam sensitivity [16].
Materials and methods. Presentation of the study area. The study area is part of a larger ensemble and covers 31cities.
Sample collection. The taking of a water sample is a delicate operation to which great care must be taken [22]. Water samples were collected during the period from January 2017 to September 2018, with two outings per week (175 samples), which consists of taking 0.5 liters of water from 20 glass bottles.
Bottles of water maintained at a temperature between 4 ° C and 6 ° C in a suitable container, the maximum time between sampling and the beginning of the analysis shall not exceed 24 h. Water was analyzed by the membrane filtration technique [19]and the incorporation technique [11].
Table 1
Number of water withdrawal carried out in the 31 cities of Algeria_
Number of City
water
withdrawal
63 Bejai'a
13 Biskra, Blida
10 Setif
09 Tebessa
07 El Bayadh
06 Bordj Bou Arreridj
05 Batna
04 Souk Ahras, Skikda, Mila, Khenchela
03 Tlemcen, Tamanrasset, Bouira, Alger
02 An Defla, Tipaza, Annaba, Constantine, Tizi
01 Ouzou, Tiaret
Chlef, Djelfa, Oum El Bouaghi, Mascara,
Saida, Bechar, Jijel, Laghouat, Guelma
Table 2
The number of microbial germs (A. hydrophila and P. aeruginosa) found during the January 2017 / September 2018 period in groundwater in Algeria
City Number of Number of
Aeromonas Pseudomonas
hydrophila UFC / 250ml of water Aeruginosa UFC / 250ml of water
Bejaia 630 547
Biskra 41 543
Blida 108 0
El Bayadh 96 20
Tlemcen 0 96
Guelma 726 0
Batna 895 0
Skikda 0 335
Tebessa 168 0
Setif 490 698
Bordj Bou 117 0
Arreridj
Alger 150 0
Souk Ahras 192 0
Mila 409 0
Tamanrasset 0 500
Annaba 90 0
Djelfa 0 27
Oum El 40 0
Bouaghi
Tiaret 450 0
Bouira 214 0
Table 3
Others microbial germs found in groundwaters in Algeria
Others bacteria found in waters Number of Bacteria UFC/250ml
Aeromonas salmonicida ssp 65
salmonicida 550
Aeromonas caviae 400
Pseudomonas putida 430
Pseudomonas alcaligenes 500
Pseudomonas fluorescens 500
Pseudomonas oryzihabitans 80
Pseudomonas luteola
Results and discussion. Microbiological quality of groundwater. The assessment of the bacteriological quality of the waters in the area was
followed by the analysis of the water harvested at 31cities during the period of January 2017 / September 2018.
The average A. hydrophila bacteria for the 31 cities during the study period is 239 ufc / 250ml. The maximum value is of the order of 895 ufc /250 ml at the Batna city. Regarding P. aeruginosa, the average is of the order of 138ufc / 250ml. The maximum value is recorded in the city of Setif (698ufc / 250ml). The number of P. aeruginosa in the cities of: Blida, Guelma, Batna, Tebessa, Bordj Bou Arreridj, Algiers, Souk Ahras, Mila, Annaba, Oum El Bouaghi, Tiaret, Bouira is around 0ufc / 250 ml. We have also noted a value of the order 0 ufc / 250 ml for A. hydrophila in the cities of: Tlemcen, Skikda, Tamanrasset, Djelfa. While Aeromonas caviae (max value: 550 ufc / 250 ml) and Pseudomonas fluorescens, Pseudomonas oryzihabitans (max value: 500 ufc / 250 ml). We also note the presence of: Pseudomonas putida, Pseudomonas alcaligenes (max value: 400 and 430 ufc / 250 ml), Pseudomonas luteola (max value: 80ufc / 250 ml), Aeromonas salmonicida ssp salmonicida (max value: 65ufc / 250 ml). The high number of these microbial germs could be explained by the poor protection of groundwater.
Conclusion. The study of the bacteriological quality of groundwater in Algeria is important especially to evaluate the risk P. aeruginosa, A. hydrophila, A.caviae. A. salmonicida ssp salmonicida, Pseudomonas putida, Pseudomonas alcaligenes, Pseudomonas fluorescens, Pseudomonas oryzihabitans and Pseudomonas luteola. Increased pH, electrical conductivity, dissolved oxygen, chloride, sodium, potassium and alkaline earth elements in groundwater promote the development of P. aeruginosa and A. hydrophila. This study challenges us to take steps to set up a monitoring system to monitor the quality of the water we consume and to prevent toxi-infections.
References
1. Glucose repression of pigment production in atypical isolates of Aeromonas salmonicida responsible for goldfish ulcer disease/K. Altmann [et al.] //J. Microbios. -1992. -Vol. 72. - P. 215-220.
2. Novel resistance to imipenem associated with an alteredpbp-4 in a pseudomonas aeruginosa clinical isolate/F. Bellido [et al.] // Journal of Antimicrobial Chemotherapy. - 1990. - P. 57-68.
3. Boutin C. Premières donnéessur la faune des puits des environs de Marrakech / C. Boutin, M. Boulanouar // Verh. Internat. Verein. Limnol. - 1984. - Vol. 22. -P. 1762-1765.
4. Immunidiagnosis of virulent strains of Aeromonas hydrophila associated with epizotic ulcerative syndrome (EUS) using a monoclonal antibody / G.A. Cartwright [et al.] // J. Fish Diseases. - 1994. - Vol. 17. - P. 123-133.
5. Falagas M.E. Colistin: the revival of polymyxins for the management of multidrug-resistant Gram-negative bacterial infections/M.E. Falagas, S.K. Kasiakou // Clin Infect Dis. - 2005. - Vol. 40. - P. 1333-41.
6. Fellrath M. Qualité des eaux. Monographie d'écologie des polluants/ M.Fellrath, D.J. Tarradellas // École Polytechnique Fédérale de Lausane. - 1984.
7. François B. Multirésistance chez Pseudomonas aeruginosa /B. François, W. Michel // J. med sci. - 2010. - Vol. 26, №11. - P. 960-968.
8. Oxa-11, an extended-spectrum variant of oxa-10 (pse-2) beta-lactamase from pseudomonas aeruginosa / L.M. Hall [et al.] //Antimicrobial Agents and Chemotherapy. - 1993. - P. 1637-44.
9. Iyobe S. Cloning and expression in enterobacteriaceae of the extended- spectrum beta-lactamase gene from a pseudomonas aeruginosa plasmid / S. Iyobe, M. Tsunoda, S. Mitsuhashi //FEMS Microbiology Letters. - 1994. - P. 175-80.
10. Jayamonah S. Effect of DO on kinetics of nitrification /S. Jayamonah, S. Ohgaki, K. Hanaki // Water Supply. - Vol. 6. - P. 141-150.
11. Microbiologie Technique/ J.N. Joffin [et al.] //Dictionnaire Des Techniques Broché. -Vol. 4. - P. 368.
12. Kreisel W. Water quality and health / W. Kreisel // Wat. Sci. Tech. - 1991. - Vol. 23. - P. 201-209.
13. Lamy B. Aeromonas et aéromonoses en médecine humaine /B. Lamy // J. Elsevier. -2012. -Vol. 12. - P. 1-7.
14. Larpent J. Mémento technique de microbio/ J. Larpent, M. Larpent-Gourgaud // Tec & Doc Lavoisier. - 1990. - Vol. 2.
15. Leminor L. Bactériologie médicale / L. Leminor, M. Véron // Flammarion Paris. -1989. - Vol. 2.
16. Beta-lactamase inhibitors are substrates for the multidrug efflux pumps of pseudomonas aeruginosa / X.Z. Li [et al.] //Antimicrobial Agents and Chemotherapy. - 1998. - P. 399-403.
17. Marshall K. C. Adhésion and growth of bacteria at surfaces in oligotrophic habitats. Can / K. C. Marshall // J. Microbiol. -1988. - Vol. 34. - P. 503-506.
18. Encapsulation of Streptococcus uberis:influence of storage and cultural condition / K. Matthews [et al.] // Vet. Microbiol. -1994. - Vol. 39. - P. 361-367.
19. Cours de microbiologie générale avec problèmes et exercices corrigés Broché /A. Meyer [et al.] // Doin France. - 2004. - P. 430.
20. Distribution de Pseudomonas aeruginosa et Aeromonas hydrophila dans les eaux de la nappe phréatique superficielle en zone équatoriale au Cameroun et relations avec quelques paramètres chimiques du milieu /M. Nola [et al.] //Revue des sciences de l'eau. - 2001. - Vol. 14, №1. - P. 4-5.
21. Infections in medical intensive care units in the United States/ M.J. Richards [et al.] // National nosocomial infections surveillance system. Crit Care Med. - 1999. -Vol. 27. -P. 887-92.
22. L'analyse de l'eau / J. Rodier [et al.] // DUNOD Paris. - 2009. - P. 1579.
23. Rossolini G.M. The aeromonas metallo-beta-lactamases: genetics, enzymology, and contribution to drug resistance / G.M. Rossolini, T. Walsh, G. Amicosante // review [34 refs].
24. Distribution of cpha or related carbapenemase-encoding genes and production of carbapene mase activity in members of the genus aeromonas / G.M. Rossolini [et al.] //Antimicrobial Agents and Chemotherapy. - 1995. - P. 346-9.
25. Roszak B. Survival stratégies of bacteria in the natural environment / B. Roszak, R. Colwell // Microbiol. Rev. - 1987. -Vol. 51. - P. 365-379.
26. Sander C. Factors controlling bacterial production in marine and freshwater sediments / C. Sander, J. Kalff// Microb. Ecol. - 1993. - Vol. 26. - P. 79-99.
27. Tuncan U. Effect of cold température on germicidal efficacy of quaternary ammonium compound, iodophor, and chlorine on Listeria / U. Tuncan // J. Food Protection. -1993. - Vol. 56. - P. 1029-1033.
28. Zn(ii) dependence of the aeromonas hydrophila ae036 metallo-beta-lactamaseactivity and stability / M.H. Valladares [et al.] //Biochemistry. - 1997. - P. 11534-11541.
29. A clinical isolate of aeromonas sobria with three chromosomally mediated inducible beta-lactamases: a cephalosporinase, a penicillinase and a third enzyme, displaying carbapenemase activity / T.R. Walsh [et al.] // Journal of Antimicrobial Chemotherapy. -1 995. - P. 271-9.
30. Wiebe J. Bacterial growth in the cold: évidence for an enhanced substrate requirement.Appl/ J. Wiebe, M. Sheldon, R. Pomeroy //Environ. Microbiol. - 1992. -Vol. 58. - P. 359-364.
31. Zhou X. Y. Role of cephalosporinase in carbapenem resistance of clinical isolates ofpseudomonas aeruginosa / X.Y. Zhou, M.D. Kitzis, L. Gutmann //Antimicrobial Agents and Chemotherapy. - 1993. - P. 1387-9.
РАСПРЕДЕЛЕНИЕ PSEUDOMONAS AERUGINOSA И AEROMONAS HYDROPHILA В
ПОДЗЕМНЫХ ВОДАХ В АЛЖИРЕ
HAFFARESSAS Y., AYAD N., BOUSSAYOUD R., MOUFFOKF. Институт Пастера Алжира, Дели-Брахим, Алжир e-mail: [email protected]
Аннотация
Микробиологическое исследование проводилось в течение одного года и девяти месяцев на подземных водах в Алжире. В период с января 2017 года по сентябрь 2018 года в 31 городе Алжира было взято 175 проб грунтовых вод. В этом исследовании использовался метод мембранной фильтрации, в результате которого микробиологическое качество грунтовых вод показало высокую частоту патогенных бактерий. Обилие Pseudomonas aeruginosa и Aeromonas hydrophila варьируется и претерпевает широкие пространственно-временные колебания. Сльное микробиологическое загрязнение воды является значительным риском для здоровья алжирского населения, регулярно употребляющего воду.
Ключевые слова: Подземные воды, P.aeruginosa, A.hydrophila,Mикробиологическое качество.
