AN ANTIBIOGRAM STUDY OF SALMONELLA AND E.COLI ISOLATED FROM BROILERS FARMS IN AL GUBA AND SHAHAT.

Abdulkarim F. H. Adam 1 , Tufahah .M .O. Atiyahullah 1 , Najwa. A. Ismaael 1 ,Osama K. Gaidan 2 Tawfig. El Tigani.Mohamed 1 and Atiyah. A. A. salih 3 . 1. Department of preventive medicine and public health Faculty of veterinary medicine. Omar ALmukhtar University,P.O.Box 919 ELBeida,Libya. 2. Department of Animal medicine and Surgery Faculty of veterinary medicine. Omar ALmukhtar University,P.O.Box 919 ELBeida,Libya. ...................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History

In the present study, the isolation of salmonella and E.coli and their antibiogram patterns in broiler farms in Alguba and Shahat were performed. Tissue samples including (liver, heart blood, spleen, exudates air sac, and intestine) were collected from 60 fresh dead or sick chickens obtained from five and three broiler farms located in Shahat and Alguba, respectively. The isolation and the identification Salmonella and E.coli were performed according to standard methods and the resistance to 13 antimicrobial agents was determined by using Kirby-Bauer disk diffusion method. In the results, E.coli and Salmonella were isolated at percentage of 21.6% and 11.6% respectively. Resistance pattern of Salmonella in descending order was respectively Amoxicillin (100%), Ampicillin(100%), Erythromycin (100%), Doxycycline (50%), Oxytetracyclin (50%), Tetracycline (50%), and Spectinomycin (25%). Whereas E.coli isolates were 100% resistant to Amoxicillin, Ampicillin, Tetracycline, Erythromycin, followed by Enrofloxacin (80%), Oxytetracyclin (80%), Streptomycin (80%), Doxycycline (80%), Sulphamethoxazole (40%), Spectinomycin (60%), and Neomycin (60%).Also in this study, all of salmonella isolates were sensitive to Gentamycin or Colistin sulphate. sensitive pattern of E.coli was recorded to Colistin, followed by Sulphamethoxazole, and Spectinomycin. Moreover, all the isolates of Salmonella and E.coli showed multiple antimicrobial resistances. It is concluded that the rational use of antibiotics broilers farming system should be applied in order to prevent the emergence of antimicrobial resistance pathogens. Salmonella have been isolated from nearly all vertebrates, and the infections with Salmonella have been recorded in both animal and human worldwide. In fact, Salmonella are responsible for pullorum diseases, fowl typhoid, and fowl paratyphoid in poultry (Hassan et al, 2014).
The growing increase in antimicrobial resistance pathogens such as Escherichia coli and Salmonella has become a critical public health concern (Hanson et al, 2002). In other word, poultry meat ranks first or second in food -related illness (Sams AR 2001). Processed raw poultry meat naturally harbors many types of bacteria and most of these bacteria are pathogenic to humans (Mead GC 2001).
The contamination can occur at several points throughout the processing operation (Ayres CP 1995). This may lead to the transfer of the resistant genes located in Salmonella and other Zoonotic bacteria from food animals to humans through consumption of contaminated food and food products

Material and Methods:-
Questionnaire:-A total of 15 questions structured were managed by the research team and administered to five and three broiler farms in Shahat and Alguba respectively. The objective of this questionnaire was to determine the most commonly used antimicrobial in broiler farms .

Isolation and Identification:-
This study was conducted on 60 fresh dead or sick chickens collected randomly from five and three broiler farms in Shahat and Alguba respectively, located in Al Jabal Alakhdar Region during the period of January to February.
Samples were aseptically collected from internal organs including liver, heart blood, spleen, exudates air sac and processed according to
Where the result indicated that the frequency of both multi-drug resistance of Salmonella and E.coli was (75%) and (100%), respectively ( Table 2).

Discussion:-
One hundred percent resistance of both isolates to B-lactam group (Amoxicillin and Ampicillin) was found and that was in agreement with (Guerra et al, 2003 ; Abd -El tawab et al, 2015). Similarly, 100% resistance to Ampicillin was recorded in Eygpt, 3% in Brazil, and 10% in Chile (Amàbile-Cuevas et al, 2010). Ampicillin resistance in Salmonella and E.coli isolates coincided with the results of (Soomro et al, 2009; Suresh et al, 2006). On the other hand, this findings were not in agreement with a study conducted by (Eissa, 2016), in camel, where isolated Salmonella showed high sensitivity to Ampicillin (100%) and Amoxicillin (100%). Interestingly, all isolates which were resistant to Ampicillin exhibited also resistance to Amoxicillin. This may be as a result of that both agents are pharmacological rela (Molla et al, 2003).
The resistance of some Salmonella and E.coli isolates to Streptomycin was (50%) and (80%), respectively. The resistance to tetracycline was also reported to be 50% in Salmonella and 100% in Ecoli. The resistance to these antimicrobial was previously demonstrated by (Castanon, 2007; Kilonzo-Nthenge et al, 2013; Phagoo and Neetoo, 2015). Resistance to tetracycline has been reported to be genetically associated with mobile plasmids or transposons (Phagoo and Neetoo, 2015). In addition, a study carried out by (Chopra and Roberts, 2001) indicated that the resistance to Tetracycline can result in inhibiting the synthesis of bacterial protein through preventing t-RNA attachment to ribosome. Furthermore, Tetracycline has been used in production animals such as poultry from one day to old broilers. Thus, the resistance developed as a result of the frequent exposure to this antibiotic during the growth period (Soomro et al, 2009). This can also be due to sub active dose and extensive use of antibiotics in poultry farms (Phagoo and Neetoo, 2015).
Macrolide antibiotics (Erythromycin) have been known for a long time for their use as therapeutic agents and growth promotion. Therefore, the resistance of some isolates to Erythromycin has been expected by investigators and may be attributed to the long term use as a growth promoter (Wanger and Cerniglia, 2005). The results also showed high multi-drug resistance of Salmonella and E.coli isolates to more than two antimicrobials. This probably due to the misusage of antibiotics administrated during the production stage (Miranda  et al, 2010). In addition, in developing countries, it is very easy for both patients and farmers to obtain antibiotics from the pharmacist without a medical supervision. The emergence of multi-drug resistance organisms has been considered to be a critical issue especially in the case of the management of diseases caused by these organisms (Tajabakhsh at al, 2015). Since it is difficult to treat infection with multi-drug resistant pathogens in compare to the susceptible one (Akbar-Anal, 2013). This resistance was mostly to for antimicrobial used extensively as feed additives or therapeutics (Tajabakhsh et al, 2015) 1283 The frequency of multi-drug resistant of Salmonella was 100%. The same result was recorded in another country, which is Nepal (Shrestha et al, 2010).  , 2005). Additionally, multidrug resistance of E.coli isolates could have resulted from the co-selection of resistance determinants, since the exposure of bacteria to one type of antibiotics may lead to resistance to other type of antibiotics without prior exposure (Miranda et al, 2010).

Conclusion:-
The study presented clear evidence on the emergence of drug resistant bacterial pathogens in veterinary field. It was found that many antibiotics have become not efficient against Salmonella and E.coli infections. Therefore, the subtherapeutic and non-therapeutic use of antibiotics in veterinary medicine should be controlled. In our study, all of salmonella and E.coli isolates were sensitive to Colistin, which might make it the drug of choice for treatment of salmonella and Ecoli infection. This study also developed the hypothesis that theses drug -resistant pathogens can transfer to the carcasses and subsequently to human via contaminated carcass processing, or meat consumption. This can lead us to another critical point at which the cost of health serving due to antibiotics resistance is expected. Food safety research should be conducted to monitor the presence of theses pathogens and reduce their impact on public health.