Veterinary Medicine International
Volume 2018 (2018), Article ID 4636121, 5 pages
https://doi.org/10.1155/2018/4636121

Research Article

Carriage Methicillin-Resistant Staphylococcus aureus in Poultry and Cattle in Northern Algeria

Saliha Bounar-Kechih,¹ Mossadak Taha Hamdi,² Hebib Aggad,³ Nacima Meguenni,⁴ and Zafer Cantekin⁵

¹Regional Veterinary Laboratory of Draa-Ben-Khedda, Tizi-Ouzou, Algeria
²HASAQ Research Laboratories, High National Veterinary School of Algiers, El-Harrach, Algiers, Algeria
³Laboratory of Hygiene and Animal Pathology, Institute of Veterinary Sciences, University of Tiaret, Algeria
⁴Department of Biochemistry Microbiology, Faculty of Biological and Agronomics Sciences, Mouloud Mammeri University, Tizi-Ouzou, Algeria
⁵Department of Microbiology, Faculty of Veterinary Medicine, Mustafa Kemal University, Serinyol, Antakya, Turkey

Correspondence should be addressed to Hebib Aggad; h_aggad@yahoo.com

Received 13 February 2018; Accepted 23 May 2018; Published 20 June 2018

Academic Editor: Francesca Mancianti

Copyright © 2018 Saliha Bounar-Kechih et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract.

Multiresistant and especially Methicillin-Resistant Staphylococcus aureus (MRSA) poses a serious public health problem that requires their immediate identification and antibiotic resistance characteristics. In order to determine antibiotic resistance S. aureus poultry and bovine origin, 8840 samples were collected from slaughterhouses in the northern region of Algeria between years 2009 and 2014. 8375 samples were from an avian origin (1875 from laying hens and 6500 from broiler chickens) and the rest was from bovine origin. Bacteriological isolation and identification were made by classical culture method and antibiotic resistance patterns were determined by disc diffusion test. The prevalence of S. aureus was 42% in laying hens, 12% in broilers, and 55% in bovine samples. The prevalence of MRSA was 57%, 50%, and 31% in laying hens, broiler chickens, and bovine, respectively. While MRSA strains isolated from poultry showed cross-resistance to aminoglycosides, fluoroquinolones, macrolides, sulphonamides, and cyclins, those isolated from bovine also revealed similar multiresistance except for sulphonamide. This high percentage of methicillin resistance and multidrug resistance in S. aureus poultry and bovine origin may have importance for human health and curing of human infections.

1. Introduction

Antibiotic resistance is prevalent in all parts of the world. Antibiotic resistance developed by bacteria is an increasingly serious threat to global public health that requires immediate actions [1]. Methicillin-resistant Staphylococcus aureus (MRSA) is a well-known pathogen of human and animals most commonly isolated from clinical cases [2]. MRSA was first identified in England in 1961 after which it emerged worldwide [3]. It was reported that MRSA clone ST80-IV, an epidemic community-acquired Methicillin-Resistant Staphylococcus aureus (C-MRSA), is widespread in the Algiers hospital and community settings [4]. Nasal carriage of multiresistant Staphylococci and MRSA in calves was detected and emphasized that they can be a cause of severe disease in humans [5]. In another study, antibiotic resistance Staphylococci isolated from poultry in the 1970s and 2006 was reported. Carriage of MRSA in healthy poultry was reported and emphasized that antibiotic resistance was higher in the recent isolates than prior isolates [6]. A high percentage of methicillin resistance in Staphylococci isolated from bovine mastitis was also reported in Algeria [7].

This study aimed to determine the occurrence of MRSA from laying hens, broilers, and bovine and their antibiotic resistance.

2. Materials and Methods

2.1. Study Area and Sampling

The samples from poultry and bovine were collected from different private farms of six different governorates located in northern Algeria (Bouira, Boumerdes, Tizi-Ouzou, Bejaia, Bordj-Bou-Arreridj, and Msila). Nasal swab samples were taken at the slaughterhouse after slaughtering. Total 8840 random samples were collected between 2009 and 2014; 8375 samples were from an avian origin (1875 were from laying hens and 6500 from broilers); and 465 samples were from bovine origin (Table 1).

Table 1: Sample distribution by year.

Poultry
Bovine

Laying hens
Broilers

2009
347
765
76

2010
306
970
124

2011
412
1327
85

2013
376
1785
78

2014
434
1653
102

Total
1875
6500
465

A sterile cotton-swab was inserted as deep as possible into the nasal cavity in order to sample collection poultry. For cattle, sterile cotton-swab was inserted into the nasal cavity at a fixed distance of 10 cm to reach the ventral meatus. Each swab sample was placed in 3 ml of a transport medium (Staphylococcus enrichment broth, Axonlab) without antibiotics and transferred in an icebox. The analysis was performed the same day at the regional veterinary laboratory of the National Institute of Veterinary Medicine (Tizi-Ouzou, Algeria).

2.2. Bacterial Isolation and Identification

Nasal swabs were streaked onto blood agar supplemented with sheep blood 7% v/v and all inoculated media were incubated at 37°C for 24 hours as described [8, 9]. The bacterial strain purification was done on nutrient agar. Bacterial strains were identified based on the morphological criteria, biochemical characters, agglutination tests (Pastorex TM Staph-Plus kit, Bio-Rad Laboratories, France), and rabbit plasma coagulase test (Bio-Rad). All identified bacterial strains were stored -20°C in Trypticase soy broth contains 20 % glycerol.

2.3. Antibiotic Resistance and Susceptibility Testing

The antibiogram was performed in the same laboratory using the disk diffusion method according Clinical Laboratory Standards Institute [10] instructions in Mueller-Hinton agar (Oxoid). S. aureus ATCC29213 Oxacillin-sensitive strain and S. aureus ATCC43300 Oxacillin resistant strain were used as control strains. Twelve different antibiotic disks were used; Oxacillin (OXA, 1μg), Penicillin G (PEN, 10UI), Cefoxitin (FOX, 30μg), Kanamycin (KAN, 30μg), Neomycin (NEO, 30μg), Gentamicin (GEN, 10μg), Tetracycline (TCY, 30μg), Trimethoprim/Sulfamethoxazole (SXT, 1.25 μg/23.75μg), Enrofloxacin (ENR, 5 μg), Erythromycin (ERY, 15μg), and Vancomycin (VAN, 30μg) (Oxoid, Hampshire, United Kingdom). Analysis of antimicrobial susceptibility test results was made by using software WHONET Version 5.6.

3. Results

Variable numbers and percentage of S. aureus were observed which were carried in laying hens, broilers, and bovines. Laying hens are shown to carry 42%, broiler chickens 12%, and bovine 55% S. aureus (Table 2).

Table 2: Distribution of S. aureus and MRSA strain.

Laying hens 
n=1875
Broiler 
n=6500
Bovine 
n=465

Nb analysed samples
1875/8840
6500/8840
465/8840

(%)
(21%)
(74%)
(5%)

Nb S. aureus
786/1875
789/6500
258/465

(%)
(42%)
(12%)
(55%)

Nb MRSA
448/786
416/789
80/258

(%)
(57%)
(53%)
(31%)

Table 3 shows percentages of significant resistance and sensitivity of poultry and bovine isolated S. aureus against a single antibiotic.

Table 3: Percentage of antibiotic resistance and sensitivity in S. aureus.

Laying hens
Broiler
Bovine

Antibiotic
Resistance
Sensitive
Resistance
Sensitive
Resistance
Sensitive

OXA
57
43
53
47
31
69

PEN
79
21
92
8
72
28

FOX
57
43
53
47
31
69

NEO
10
90
10
90
11
89

KAN
0
100
67
33
12
88

GEN
3
97
3
97
0
100

SXT
21
79
28
72
0
100

TCY
44
56
74
26
26
74

ENR
21
79
45
55
87
13

ERY
55
45
45
55
51
49

VAN
0
100
0
100
0
100

Oxacillin (OXA), Penicillin (PEN), Cefoxitin (FOX), Neomycin (NEO), Kanamycin (KAN), Gentamicin (GEN), Trimethoprim/Sulfamethoxazole (SXT), Tetracycline (TCY), Enrofloxacin (ENR), Erythromycin (ERY), and Vancomycin (VAN).

A total of 57 % of Oxacillin resistant MRSA was observed in laying hens, 53% in broilers, and 31% in bovine. Likewise, 79 % Penicillin-resistant S. aureus was observed in laying hens, 92% in broilers, and 72 % in bovine. The resistance of S. aureus to other antibiotics is significantly mentioned in Table 3.

Interestingly and expectedly all bacterial isolates were simultaneously sensitive to Gentamicin (Table 3).

Further, multiple antibiotic cross-resistance patterns of MRSA identified in laying hens, broilers, and bovines were analyzed. Different nine antibiotics belonging to six different families were used, all including Oxacillin. Results shows MRSA strain identified in laying hens, broilers, and bovines that cross-tolerated multiple antibiotics together with Oxacillin (Tables 4, 5, and 6).

Table 4: Antibiotic resistance patterns of MRSA in hens (n=448).

Resistance patterns
Number of antibiotics
MRSA

OXA-PEN-FOX-ERY
3
148

OXA-PEN-FOX-TCY
4
65

OXA-PEN-FOX-ERY-TCY
5
23

OXA-PEN-FOX-SXT-TCY-ENR-ERY
7
21

OXA-PEN-FOX
3
17

OXA-PEN-FOX-TCY-ENR
5
17

OXA-PEN-FOX-SXT-TCY-ENR
6
16

OXA-PEN-FOX-NEO-SXT-TCY-ENR
7
14

OXA-PEN-FOX-NEO-SXT-TCY-ENR
7
14

OXA-PEN-FOX-SXT-TCY
5
11

OXA-PEN-FOX-NEO-TCY
5
11

OXA-PEN-FOX-SXT-TCY-ENR
6
11

OXA-PEN-FOX-SXT-TCY
5
10

OXA-FOX-TCY
3
7

OXA-PEN-FOX-SXT
4
7

OXA-PEN-FOX-TCY-GEN
5
7

OXA-PEN-FOX-TCY-NEO
4
7

OXA-PEN-FOX-ENR
4
7

OXA-PEN-FOX-SXT-ENR
5
7

OXA-PEN-FOX-NEO-SXT-TCY-ERY
7
7

OXA-PEN-FOX-TCY-ENR-ERY
6
7

OXA-PEN-FOX-NEO-GEN-SXT-TCY
7
7

OXA-PEN-FOX-NEO-GEN-SXT-TCY-ENR
8
7

Table 5: Antibiotic resistance patterns of MRSA isolated from broiler (n=416).

Antibiotic profiles
Number of antibiotics
MRSA

OXA-FOX-TCY
3
82

OXA-PEN-FOX-TCY
4
64

OXA-PEN-FOX-SXT
4
58

OXA-PEN-FOX
3
68

OXA-PEN-FOX-TCY
4
28

OXA-PEN-FOX-SXT-TCY
5
16

OXA-PEN-FOX-ENR-ERY
5
8

OXA-PEN-FOX-SXT-ENR
5
8

OXA-PEN- FOX-NEO-GEN-SXT-TCY
7
8

OXA-PEN-FOX-NEO-GEN-SXT-TCY-ENR
8
8

OXA-PEN-FOX-TCY-ENR-ERY
6
8

OXA-PEN-FOX-SXT-ENR-ERY
6
8

OXA-PEN-FOX-GEN-TCY-ERY
6
8

OXA-PEN-FOX-NEO-TCY
5
8

OXA-PEN-FOX-NEO-SXT-TCY-ENR
7
8

OXA-PEN-FOX-TCY-ENR
5
6

OXA-PEN-FOX-NEO-SXT-TCY-ENR
7
6

OXA-PEN-FOX-NEO-TCY
5
6

OXA-PEN-FOX-SXT-TCY-ENR
6
6

OXA-PEN-FOX-SXT-TCY
5
4

Table 6: Antibiotic resistance patterns of MRSA isolated from bovine (n=80).

Antibiotic profiles
Number of antibiotics
MRSA

OXA-PEN-FOX
3
34

OXA-PEN-FOX-TCY
4
15

OXA-PEN-FOX-ERY
4
10

OXA-PEN-FOX-NEO-KAN-TCY
6
8

OXA-PEN-FOX-ENR
4
6

OXA-PEN-FOX-ERY-ENR
5
4

OXA-PEN-FOX-NEO-KAN-TCY-ENR
7
2

OXA-PEN-FOX-ENR-ERY
5
1

More importantly and comparatively in OXA-PEN-FOX-ERY combination, MRSA showed greater cross-resistivity in laying hens 33% (148/448) (Table 4) than in bovine 12% (10/80) (Table 6). Also cross-resistance of MRSA towards OXA-PEN-FOX-TCY combination was observed slightly less in broiler 16% (64/416) (Table 5) than in bovine 19% (15/80) as mentioned in Table 6 although highest noncomparative cross-resistance in broiler MRSA was 20% (82/448) towards OXA-FOX-TCY combination (Table 5) and 43% in bovine MRSA against OXA-PEN-FOX combination (Table 6).

4. Discussion

Globally Staphylococcus aureus bacterial pathogens are becoming resistant towards multiple antibiotics. Methicillin-resistant Staphylococcus aureus (MRSA) is continuously failing to be sensitive to β-lactam antibiotics that have caused severe problems in human and veterinary medicines. Like other countries of the world, Algeria is also facing emerging dangers posed by S. aureus and MRSA and knowledge is quite limited on the prevalence of such bacterial pathogens in Algeria compared to the world because of the lack of advanced monitoring system and tools. Although the total number of samples we analyzed from laying hens, broilers and bovine were not equal; it was observed that laying hens and broiler isolated MRSA strains were more resistant towards single or multiple antibiotic combinations; therefore they were cross-resistant to many other antibiotic families other than β-lactam antibiotics. A study carried out in eastern Algeria also showed that MRSA isolates in a hospital were resistant towards aminoglycosides, macrolides, fluoroquinolones, and tetracycline in many combinations. In another study in North-west of Algeria, different level of Vancomycin resistance of MRSA had been reported which was 54 % in poultry and 1.8 % in human [11]. In Algeria, selling antibiotics is totally uncontrolled what means abusive and indiscriminate use in human but also in the animal. Indeed, 89.09 % of the milk from 4 governorates has been found positive to antibiotics residues [12]. Moreover, 29 % of the examined milk samples produced in western Algeria contained antibacterial residues [13].

A study conducted in Egypt indicates a strong correlation between methicillin resistance and cross-resistance to β-lactam antibiotics [14]. Modification in Penicillin-binding protein (PLP2a) induces inactivity of Penicillin and other β-lactam antibiotics that develop cross-resistance of multiple antibiotics. Confirmed hospital report on MRSA in the human patient in Algeria increased our interest to investigate the prevalence of S. aureus and MRSA in poultry and bovine in a particular area to link with the human patient who lived in the close vicinity of such poultry and bovine animals. Of note, this antibiotic resistance data, especially on poultry, is strongly supported by previous studies like in Belgium [6] which have shown that S. aureus was resistant to tetracycline, macrolide, and cotrimoxazole greatly in broiler chickens. Our data on cross-resistance of MRSA in broiler chickens, towards eight antibiotics from six different families, is further supported by the previous study [5]. Multiple antibiotics cross-resistance in MRSA in Germany originated in broiler chicken and turkey birds [15] support our findings. However, our results show a higher prevalence of MRSA (53%) in broiler chickens than those reported which showed a prevalence of MRSA in broiler chickens (37.2%) and in turkey birds (35%). The previous study also shows the prevalence of S. aureus strains in poultry in Korea and Japan [5]. 44% prevalence of MRSA was also reported in poultry [14]. The prevalence of MRSA infections in African countries is relatively high in Nigeria, Kenya, and Cameroon, which is up to 30% and less than 10% in Tunisia [16]. In Algeria, the frequency of MRSA infection was reported to be 14 % in 2001 and recently Algerian hospitals have demonstrated a significant increase in MRSA that is up to 40% [4, 17]. Interestingly, MRSA incidence in French hospital is shown to be reduced [18]. Many studies suggest that farm animals show increasing trends of MRSA which is slightly lower in human that indicates farm animals are exposed to huge multiple antibiotics with little medical care than the humans. The emergence of MRSA came in human from swine and mecA gene in MRSA is predominant host restriction factors [19]. The ST398 clone has been found in different animal species including poultry and transmission of MRSA from mastitis cow was identified in a human who was working with these cows [20, 21]. Coagulase positive S. aureus isolates in live and slaughtered chickens were shown to be 100 % resistance towards tetracycline, Penicillin, and erythromycin and were additionally resistant to other antibiotics including Vancomycin (46.2 %) [22]. New MRSA clones and mec homologs among animals of the food chain and human in close contacts have been also reported [23]. The result of this study shows 57% MRSA in S. aureus isolated from laying hens, 53% in broiler chickens and 31% in bovine that indicates an increasing trend.

The main finding of this study clearly indicates that S. aureus and MRSA isolated from avian origin were more resistant to β-lactam antibiotics (Penicillin and Oxacillin) than bovine. This result also suggests MRSA transmission from poultry to bovine and humans that may be difficult to treat these days. In this respect, they may be a potential risk for human health.

Knowing the lack of deep research in Algeria, molecular studies are needed in order to understand the cross transmission from animal to human or from human to animals.

Data Availability

The data used to support the findings of this study are available from the corresponding author upon request.

Conflicts of Interest

There are no conflicts of interest in this research project.

Acknowledgments

The authors are grateful to Pr. K. Rahal, Head of Bacteriology and Medicine Department (Pasteur Institute, Algiers), for her expertized knowledge on bacteriology. Thanks are due to Dr. M. Intakhab Alam and Dr. Yaşar Ergün for critical reading of this manuscript.

References

WHO, World Health Organization, 2015, http://www.who.int/mediacentre/factsheets/fs194/en/.

D. Persoons, S. Van Hoorebeke, K. Hermans et al., “Methicillin-resistant Staphylococcus aureus in poultry,” Emerging Infectious Diseases, vol. 15, no. 3, pp. 452-453, 2009.

A. C. Fluit, “Livestock-associated Staphylococcus aureus,” Clinical Microbiology and Infection, vol. 18, no. 8, pp. 735–744, 2012.

K. Antri, N. Rouzic, O. Dauwalder et al., “High prevalence of methicillin-resistant Staphylococcus aureus clone ST80-IV in hospital and community settings in Algiers,” Clinical Microbiology and Infection, vol. 17, no. 4, pp. 526–532, 2011.

M. Haenni, P. Châtre, S. Boisset et al., “Staphylococcal nasal carriage in calves: Multiresistant Staphylococcus sciuri and immune evasion cluster (IEC) genes in methicillin-resistant Staphylococcus aureus ST398,” Journal of Antimicrobial Chemotherapy, vol. 66, no. 8, Article ID dkr196, pp. 1927-1928, 2011.

M. Nemati, K. Hermans, U. Lipinska et al., “Antimicrobial resistance of old and recent Staphylococcus aureus isolates from poultry: first detection of livestock-associated methicillin-resistant strain ST398,” Antimicrobial Agents and Chemotherapy, vol. 52, no. 10, pp. 3817–3819, 2008.

R. Saidi, Z. Cantekin, D. Khelef, Y. Ergün, H. Solmaz, and R. Kadi, “Antibiotic susceptibility and molecular identification of antibiotic resistance genes of staphylococci isolated from bovine mastitis in Algeria,” Kafkas Universitesi Veteriner Fakultesi Dergisi, vol. 21, no. 4, pp. 513–520, 2015.

P. J. Quinn, M. E. Carter, B. K. Markey, and G. R. Carter, Clinical Veterinary Microbiology, Mosby-Year Book Europe Limited, Lynton House, London, UK, 1994.

P. Singleton, Bacteriology for Medicine, Biology and Biotechnology "Course", Dunod Paris, Paris, France, 6th edition, 2005.

CLSI, Clinical and Laboratory Standards Institute, Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Third Informational Supplement, Clinical and Laboratory Standards Institute, Wayne, Pa, USA, 2013.

S. A. Rebiahi, D. E. Abdelouahid, M. Rahmoun, S. Abdelali, and H. Azzaoui, “Emergence of vancomycin-resistant Staphylococcus aureus identified in the Tlemcen university hospital (North-West Algeria),” Médecine et Maladies Infectieuses, vol. 41, no. 12, pp. 646–651, 2011.

D. Tarzaali, A. Dechicha, S. Gharbi, M. K. Bouaissa, N. Yamnaine, and D. Guetarni, Recherche des résidus des tétracyclines et des bêta-lactamines dans le lait cru par le MRL Test (ROSA TEST) à Blida, Algérie. In 6e Journées scientifiques vétérinaires sur le médicament vétérinaire: nouvelles approches thérapeutiques et impact sur la santé publique, École nationale vétérinaire, Algérie, 2008.

H. Aggad, F. Mahouz, Y. A. Ammar, and M. Kihal, “Assessment of milk hygienic quality in Western Algeria,” Revue de Médecine Vétérinaire, vol. 160, no. 12, pp. 590–595, 2009.

M. Karmi, “Prevalence of methicillin-resistant Staphylococcus aureus in poultry meat in Qena, Egypt,” Veterinary World, vol. 6, no. 10, pp. 711–715, 2013.

A. T. Febler, K. Kadlec, M. Hassel et al., “Characterization of methicillin-resistant Staphylococcus aureus isolates from food and food products of poultry origin in Germany,” Applied and Environmental Microbiology, vol. 77, no. 20, pp. 7151–7157, 2011.

C. Kesah, S. B. Redjeb, T. O. Odugbemi et al., “Prevalence of methicillin-resistant Staphylococcus aureus in eight African hospitals and Malta,” Clinical Microbiology and Infection, vol. 9, no. 2, pp. 153–156, 2003.

N. Ramdani-Bouguessa, M. Bes, H. Meugnier et al., “Detection of methicillin-resistant Staphylococcus aureus strains resistant to multiple antibiotics and carrying the Panton-Valentine leukocidin genes in an Algiers hospital,” Antimicrobial Agents and Chemotherapy, vol. 50, no. 3, pp. 1083–1085, 2006.

P. Martres, M. Thibault, and F. Lémann, “Significant decrease of rate and incidence of methicillin resistant Staphylococcus aureus in a French general hospital between 1999 and 2001,” Pathologie Biologie, vol. 51, no. 8-9, pp. 474–478, 2003.

M. Haenni, E. Jouy, JY. Madec, and F. Laurent, “Methicillin-resistant Staphylococcus aureus (MRSA): a sharing between man and animal,” Epidemiol Bull, vol. 53, pp. 40–42, 2012.

J. P. Ganiere, “Health status of domestic fauna: zoonotic infectious diseases,” in Proceedings of the XXXVII symposium of the National Institute of medicine agricultural.Zoonoses New, Congress Vinci Centre, Tours, France, 2010.

É. Juhász-Kaszanyitzky, S. Jánosi, P. Somogyi et al., “MRSA transmission between cows and humans,” Emerging Infectious Diseases, vol. 13, no. 4, pp. 630–632, 2007.

O. Otalu Jr., K. Junaidu, O. E. Chukwudi, and U. V. Jarlath, “Multi-drug resistant coagulase positive Staphylococcus aureus from live and slaughtered chickens in Zaria, Nigeria,” International Journal of Poultry Science, vol. 10, no. 11, pp. 871–875, 2011.

E. Petinaki and I. Spiliopoulou, “Methicillin-resistant Staphylococcus aureus among companion and food-chain animals: Impact of human contacts,” Clinical Microbiology and Infection, vol. 18, no. 7, pp. 626–634, 2012.