ANTIMICROBIAL RESISTANCE AUSTRALIAN BROILERS SUMMARY

ANTIMICROBIAL RESISTANCE IN AUSTRALIAN BROILERS SUMMARY REPORT PREPARED FOR AUSTRALIAN CHICKEN MEAT FEDERATION JUNE 2009 Stephen Page Advanced Veterinary Therapeutics PO Box 345 Berry NSW 2535 AUSTRALIA spw@advet.com.au

TABLE OF CONTENTS INTRODUCTION 3 KEY MESSAGES 4 SUMMARY OF ANTIMICROBIAL RESISTANCE SURVEILLANCE REPORTS 8 KEY FINDINGS FROM ANTIMICROBIAL RESISTANCE SURVEILLANCE REPORTS 10 SUMMARY OF POULTRY CAMPYLOBACTER RESISTANCE LITERATURE 15 SUMMARY OF POULTRY SALMONELLA RESISTANCE LITERATURE 18 SUMMARY OF POULTRY ESCHERICHIA COLI RESISTANCE LITERATURE 25 SUMMARY OF POULTRY ENTEROCOCCUS SPP RESISTANCE LITERATURE 28 AMR IN POULTRY: CONSOLIDATION OF COUNTRY REPORTS 30 JUDICIOUS USE OF ANTIMICROBIAL AGENTS 34 RISK MANAGEMENT 35 ANTIMICROBIAL AGENTS FOR USE IN POULTRY: CONTROLS AND PROCESSES 37 POULTRY VACCINES 45 BROILER CHICKEN ANTIMICROBIAL AGENTS 49 APPROVED USES OF ANTIMICROBIAL AGENTS IN POULTRY 52 SCHEDULING STATUS OF ANTIMICROBIAL AGENTS APPROVED FOR USE IN POULTRY 57 Antimicrobial Resistance in Australian Broilers: Summary Page 2 of 57

INTRODUCTION Antimicrobial resistance in foodborne bacteria and the implications for possible adverse effects on public health have been a major focus of national and international attention for more than a decade. The current report was commissioned by the Australian Chicken Meat Federation with the following objectives: OBJECTIVE 1 To assess the current public health importance of antimicrobial resistance in bacteria present in poultry OBJECTIVE 2 To describe current practices from farm to fork that reduce the public health impact of bacteria with antimicrobial resistance OBJECTIVE 3 OBJECTIVE 4 To observe the impact of antimicrobial administration on animal health and welfare To propose a framework of communication about current practices that is supported by data After a comprehensive review of nearly 2,000 publications it was clear that the poultry industry in Australia has a unique position with no resistance amongst Campylobacter, E coli and Salmonella to the fluoroquinolone class of antimicrobial agents and no cephalosporin resistance in E coli and Salmonella. The uniquely low levels of resistance appear to reflect a conservative approach to registration of new antimicrobial agents, high standards of poultry production and judicious selection of use of antimicrobial agents by veterinarians. Antimicrobial Resistance in Australian Broilers: Summary Page 3 of 57

KEY MESSAGES OBJECTIVE 1 AUSTRALIAN SITUATION To assess the current public health importance of antimicrobial resistance in bacteria present in poultry (what s the reality?) No confirmed cases of harm to humans in Australia associated with exposure to bacteria of poultry origin with antimicrobial resistance Results of a pilot surveillance study of key bacterial species isolated directly from animals and undertaken on behalf of DAFF led to a conclusion by the NHMRC that control measures are successful and any impact on human health resulting from AMR is likely to be small. CAMPYLOBACTER Fluoroquinolones have never been registered in Australia for use in food production animals CAMPYLOBACTER FQ resistance in Australian Campylobacter isolates from humans usually associated with recent overseas travel (for example, FQ resistance in Thailand Campylobacter isolates reported at 77%) CAMPYLOBACTER No significant AMR was observed in the Australian retail food survey. No FQ resistance was detected and only 3.3% of isolates resistant to erythromycin, 1.7% resistant to tetracycline, and 1.7% resistant to clindamycin. CAMPYLOBACTER A literature review found that Australia was one of only 4 countries with all isolates of campylobacter susceptible to the FQs E COLI Despite widespread consumption of poultry meat, it has rarely been implicated as a source of food-borne exposure in pathogenic E. coli cases. FSANZ (2005) found that there has [been] no documented case of food-borne illness due to E. coli associated with consumption of poultry meat in Australia. E COLI Following a thorough scientific assessment of the public health and safety of poultry meat in Australia FSANZ (2005) concluded that generic [commensal] E. coli is often present on raw poultry meat, and is associated with faecal contamination during processing. However, human pathogenic strains such as EHEC have rarely been isolated from poultry. E COLI Frequently identified AMR in Australian broiler isolates was observed to the aminopenicillin ampicillin (38%), the aminoglycoside streptomycin (19%), tetracycline (47%) and the combination of sulphonamide and trimethoprim (22%) E COLI FSA study of AMR in Australian food found no evidence of resistance to the FQ and 3GC families of antimicrobial agents in E coli isolates from poultry E COLI Importantly, there were no reports of FQ or ESBL resistance amongst E coli isolates from Australia. E COLI In Australia pathogenic E. coli are considered a minimal public health and safety risk in regards to consumption of poultry meat, despite the relative severity of illness. ENTEROCOCCI Bell et al (1998a) On the basis of our findings the epidemiology of VRE in Australia appears to be different from that in either the United States or Europe. ENTEROCOCCI Christiansen et al (2007) In the 2005 Australian survey of human clinical isolates, Enterococcus faecalis (1,987 strains) and E. faecium (180 strains) made up 98.6% of the 2,197 isolates tested. Ampicillin resistance was common (77%) in E. faecium, but rare still in E. faecalis (0.2%). Resistance to vancomycin was 7.2% in E. faecium and 0.2% in E. faecalis; the vanb gene was detected in all vancomycin-resistant isolates. ENTEROCOCCI Reassuringly, the Australian retail food survey did not detect evidence of resistance to high level gentamicin or vancomycin (known as VRE), unlike the situation in Europe where VRE are still isolated, though at low levels (less than 6% in E faecium) ENTEROCOCCI The vanb gene has never been detected in food (EAGAR Factsheet). SALMONELLA In Australia there is no multiresistant Salmonella Typhimurium DT104 in broiler chickens SALMONELLA ESBL resistance not established in Salmonella isolates from Australian poultry SALMONELLA Fluoroquinolone resistance not reported in Salmonella isolates from Australian poultry SALMONELLA Multiple resistance in 2008 for broiler isolates was found to be 3.7% (much of Antimicrobial Resistance in Australian Broilers: Summary Page 4 of 57

which is associated with S Sofia) and can be compared with multiple resistance levels of 10.6% for bovine and 30.4% for porcine isolates. SALMONELLA No significant presence of Salmonella Enteritidis in broiler chickens in Australia SALMONELLA Of 10,248 salmonella isolates from broilers assessed by the Australian Salmonella Reference Centre for ciprofloxacin resistance between 2001 and 2008 inclusive, no isolates have displayed resistance. SALMONELLA Of 18,373 broiler isolates tested from 2005 to 2008, only 47 (0.26%) Enteritidis have been identified and all originated in Queensland, 18 of which were phage type 26 and none the epidemic phage type 4 which is a significant food safety concern outside Australia, especially in eggs and egg products. SALMONELLA Of 9,172 salmonella isolates from broilers assessed for cefotaxime resistance between 2002 and 2008 inclusive, only one isolate (serovar Saintpaul isolated in 2007) displayed phenotypic resistance SALMONELLA S. Sofia is the most commonly isolated Salmonella serovar from chickens in Australia and is regarded as having relatively low virulence to humans. It has a high level of resistance to Streptomycin and high level of multiresistance but is rarely isolated from humans. SALMONELLA Salmonellosis is one of the most frequently reported food-borne illnesses in Australia SALMONELLA Survey of Retail Food. Food Science Australia 2009: absence of resistance in isolates to the quinolone / fluoroquinolone (FQ) and third generation cephalosporin (3GC) classes of antimicrobial agents SALMONELLA The Australian Salmonella Reference Centre tests the resistance of isolates to a panel of 11 antimicrobial agents. Resistance to all agents is low or absent and multiple resistance is present at a low frequency. There is also a trend for reduced levels of resistance in the time period from 2001 to 2008 OBJECTIVE 1 GLOBAL SITUATION To assess the current public health importance of antimicrobial resistance in bacteria present in poultry (what s the reality?) CAMPYLOBACTER 22 countries reported ML resistance, with 13 observing <10% of isolates with ML R (including Australia) CAMPYLOBACTER Almost all surveys conducted outside Australia reveal a situation with high levels of AMR, including widespread resistance to the FQ class of antimicrobial agent in Europe (but not yet identified in Canada and the USA, though significant levels of resistance to nalidixic acid have been observed I the USA and this resistance is a common precursor to FQ resistance) CAMPYLOBACTER Fluoroquinolone sensitivity is now a rare phenomenon but present in Australia and in very few other countries. CAMPYLOBACTER Reports from 34 countries describe the presence of FQR, 12 of which had >50% of isolates with FQR and up to 88% isolates with FQR CAMPYLOBACTER Resistance by Campylobacter jejuni and C coli to the fluoroquinolone (FQ) class of antimicrobial agent is widespread throughout the world, in some countries reaching a prevalence of 80% amongst isolates from poultry E COLI A literature review found a number of cases where antimicrobial resistance in E coli was shown to be present only in imported meat, emphasising the necessity and potential value of border controls. E COLI Extended spectrum beta-lactamase (ESBL) resistance (frequently manifested as resistance to the cephalosporins, including ceftiofur) was reported in 14 countries. E COLI Resistance to the quinolone (for example nalidixic acid) and fluoroquinolone (FQ) (for example ciprofloxacin and enrofloxacin) class of antimicrobial agents was common and widespread and reported from 27 countries with up to 76% of isolates demonstrating FQ resistance. E COLI Resistance to two or more antimicrobial agents was very common and reported from 43 countries. E COLI Should be noted that E coli recovered from poultry are predominantly (almost solely) innocuous commensal bacteria they are rarely if ever the virulent disease causing types of E coli. E COLI The antimicrobial resistance patterns of E coli recovered from well birds, sick birds and raw Antimicrobial Resistance in Australian Broilers: Summary Page 5 of 57

meat was reported and reviewed in 156 publications from 50 countries. E COLI When examining AMR surveillance reports from Europe and North America it is clear that FQ and 3GC resistance has emerged. For example, CIPARS reported that among chicken abattoir generic E. coli, resistance to ceftiofur (3GC) was observed in 21% of isolates. Lower frequencies of 3GC were isolated in Europe. However, FQ resistance was widespread amongst E coli isolates from poultry in Europe, with 53% of Spanish isolates and 30% of Dutch isolates demonstrating resistance. ENTEROCOCCI Enterococcus spp display a broad spectrum of resistances, but the public health importance of resistance in isolates form animals remains uncertain with a growing number of studies suggesting that host specificity may be more important than previously appreciated. ENTEROCOCCI Oh et al (2007) VREF isolates exhibit distinct phenotypic and genotypic traits according to their origins, which suggests that no evidence exists to substantiate the clonal spread or transfer of vancomycin resistance determinants between humans and poultry ENTEROCOCCI The enterococci are well known for accumulating resistance to a variety of antimicrobial agents SALMONELLA A review of the literature revealed reports of extra-spectrum beta-lactamase resistance (EBSL) in 15 countries and quinolone / fluoroquinolone resistance in 31 countries SALMONELLA Across Europe resistance to nalidixic acid was remarkably high (range 13.2% - 89.6%) and in the Netherlands resistance to the FQ ciprofloxacin was reported at 34%. SALMONELLA By contrast high frequencies of resistance are common findings in most surveillance programs outside Australia. SALMONELLA Resistance to the 3GC ceftiofur was not reported in Europe but was not uncommonly isolated in Canada and the USA, where 19.1% of isolates were resistant. OBJECTIVE 2 To describe current practices from farm to fork that reduce the public health impact of bacteria with antimicrobial resistance JUDICIOUS USE Prudent use is defined by the World Health Organisation as usage of antimicrobials which maximizes therapeutic effect and minimizes the development of antimicrobial resistance (WHO 2000). ALTERNATIVES: VACCINES There are 46 products approved for use by the APVMA to prevent 17 diseases, including diseases caused by Bacteria (4 disease agents); Mycoplasma (2 disease agents); Protozoa (1 disease agent, but 8 species); Viruses (10 disease agents) ALTERNATIVES: VACCINES Vaccines to prevent disease in poultry production are essential tools and widely used. ANTIMICROBIAL AGENTS 155 antimicrobial products approved for use in broiler chickens: 95 antibacterial products (17 antibacterial agents) and 60 antiprotozoal products (14 antiprotozoal agents) ANTIMICROBIAL AGENTS Age of antibacterial compounds ranges from 1940 to 1973 with an average time since discovery of 52 years ANTIMICROBIAL AGENTS All antimicrobial agents in use in poultry satisfy the strict regulatory requirements of the APVMA and also satisfy the recommendation of WHO and OIE for use of critically important antimicrobial agents ANTIMICROBIAL AGENTS No new antimicrobial agents are used in broiler production ANTIMICROBIAL AGENTS Of the 17 antibacterial agents approved for use in poultry, 14 are only available on prescription from a veterinarian (Schedule 4). 3 agents (avilamycin, flavophospholipol, tylosin) are available over the counter, only one of which (tylosin) belongs to a family used in humans (macrolides or MLs). Currently the MLs are under review by the APVMA and are expected to be reclassified as S4 JUDICIOUS USE Antimicrobial agents were prescribed sparingly and generally only with confirmed or high index of suspicion (based on flock history, property history and clinical signs) of primary or secondary bacterial infection. JUDICIOUS USE In many flocks it was considered usual that no outbreaks of bacterial disease would be encountered during grow outs. JUDICIOUS USE In support of the apparently high standards of judicious use of antimicrobial agents by veterinarians involved in poultry health, recent surveys of retail poultry have identified low levels or no resistance to most antimicrobial agents tested. Antimicrobial Resistance in Australian Broilers: Summary Page 6 of 57

JUDICIOUS USE Interviews with poultry veterinarians indicated that the principles of judicious use were widely recognised and understood and underpinned current Australian prescribing practices. JUDICIOUS USE Off label use of antimicrobial agents in broiler production was not reported. JUDICIOUS USE Responses to treatment were closely monitored. JUDICIOUS USE The Federation of Veterinarians of Europe (FVE 1999) adds that Prudent use is an integral part of good veterinary practices. It is an attitude to maximise therapeutic efficacy and minimise selection of resistant micro-organisms. Prudent use principles are a guide for optimal use of antibiotics. They should not be interpreted so restrictively as to replace professional judgement of practitioners or to compromise animal health or welfare. In all cases, animals should receive prompt and effective treatment as deemed necessary by the prescribing or supervising veterinarian JUDICIOUS USE There was a strong emphasis on ensuring high standards of hygiene, biosecurity and infection control to reduce the likelihood of any presence of bacterial disease. RISK MANAGEMENT The 16 components of AMR risk management described by FAO/WHO/OIE (2008) [Joint FAO/WHO/OIE Expert Meeting on Critically Important Antimicrobials. Report of a meeting held in FAO, Rome, Italy, 26 30 November 2007., FAO, Rome, Italy, and WHO, Geneva, Switzerland] are already standard and integral components of Australian poultry production. High health flocks arising from: sophisticated housing; advanced biosecurity; strategic use of vaccination to prevent disease and maintain high levels of health; precision diets; superior husbandry Veterinary management of preventative flock health programs Judicious use of antimicrobial agents Use of alternatives to antimicrobial agents (vaccines, acidulents, probiotics) World class standards of processing OBJECTIVE 3 To observe the impact of antimicrobial administration on animal health and welfare Support of high health status Reductions in morbidity and mortality World class growth rates OBJECTIVE 4 To propose a framework of communication about current practices that is supported by data AMR surveillance portrays excellent (but not perfect) picture Role of the veterinarian in support of health and welfare Opportunities for improvement: data collection (pooled results of in-house AMR surveillance; pooled results of antibiotic use; need common definition of antibiotic ) and antimicrobial stewardship Antimicrobial Resistance in Australian Broilers: Summary Page 7 of 57

SUMMARY OF ANTIMICROBIAL RESISTANCE SURVEILLANCE REPORTS Formal programs for the surveillance of antimicrobial resistance (AMR) in isolates from broilers and other livestock species have been introduced in a number of countries. Reports from the 11 countries or regions identified in the following table were reviewed and information on the AMR status of isolates of Salmonella, Escherichia coli, Campylobacter jejuni and Enterococcus (faecium and faecalis) obtained from food or gut samples was assessed and compared with the results of a recent survey of retail food in Australia (Food Science Australia 2009). COUNTRY AMR MONITORING PROGRAMME Canada CIPARS Canadian Integrated Program for Antimicrobial Resistance Surveillance Denmark DANMAP Use of antimicrobial agents and occurrence of antimicrobial resistance in bacteria from food animals, foods and humans in Denmark European Community [Contributions of data as available from 25 member EFSA The Community Summary Report on Trends and Sources of Zoonoses, states (Austria, Belgium, Cyprus, Czech Zoonotic Agents, Antimicrobial Republic, Denmark, Estonia, Finland, Resistance and Foodborne Outbreaks France, Germany, Greece, Hungary, in the European Union Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Poland, Portugal, Slovakia, Slovenia, Spain, Sweden, United Kingdom) with variable contributions from 5 Non member states (Bulgaria, Norway, Romania, Switzerland, Liechtenstein and Iceland)] European Community de Jong et al 2009 Finland FINRES-Vet Finnish Veterinary Antimicrobial Resistance Monitoring and Consumption of Antimicrobial Agents Italy ITAVARM Italian Veterinary Antimicrobial Resistance Monitoring Japan JVARM Japanese Veterinary Antimicrobial Resistance Monitoring Program Netherlands MARAN Monitoring of Antimicrobial Resistance and Antibiotic Usage in Animals in the Netherlands In 2005 Norway NORM Usage of Antimicrobial Agents and Occurrence of Antimicrobial Resistance in Norway Spain VAV Veterinary monitoring of antimicrobial resistance in Spain Sweden SVARM Swedish Veterinary Antimicrobial Resistance Monitoring USA NARMS USA National Antimicrobial Resistance Monitoring System SALMONELLA spp The frequency of isolates from chickens sampled at retail level in Australia (Food Science Australia 2009) with resistance to antimicrobial agents was generally very low (0-5% of isolates with resistance), with the exception of resistance to tetracycline which was identified in 16% of isolates. However, tetracyclines are not used in the treatment of Salmonella infections in humans or Antimicrobial Resistance in Australian Broilers: Summary Page 8 of 57

livestock so the presence of resistance does not reflect compromise of effective treatment, but does provide an index of resistance selection and dissemination occurring in broilers or the environment to which they are exposed. Of major importance was the absence of resistance in isolates to the fluoroquinolone (FQ) and third generation cephalosporin (3GC) classes of antimicrobial agents. By contrast with the situation in Australia, high frequencies of resistance are common findings in most surveillance programs as summarised in the Tables below. For example, in Europe (EFSA 2007) the highest proportions of resistant isolates were observed for nalidixic acid (average of reporting countries 12.3%), streptomycin (26.4%) and tetracycline (27.4%). Of greatest interest and potential concern from a pubic health perspective is the emergence and dissemination of resistance to the FQ and 3GC classes of antimicrobial agents. Across Europe resistance to nalidixic acid was remarkably high (range 13.2% - 89.6%) and in the Netherlands resistance to the FQ ciprofloxacin was reported at 34%. Resistance to the 3GC ceftiofur was not reported in Europe but was not uncommonly isolated in Canada and the USA, where 19.1% of isolates were resistant. E COLI The FSA study of AMR in Australian food found no evidence of resistance to the FQ and 3GC families of antimicrobial agents in E coli isolates from poultry. Frequently identified AMRs were observed to the aminopenicillin ampicillin (38%), the aminoglycoside streptomycin (19%), tetracycline (47%) and the combination of sulphonamide and trimethoprim (22%). When examining AMR surveillance reports from Europe and North America it is clear that FQ and 3GC resistance has emerged. For example, CIPARS (2005) reported that among chicken abattoir generic E. coli, resistance to ceftiofur (a third generation cephalosporin) was observed in 21% (45/218) of isolates. Lower frequencies of 3GC were isolated in Europe. However, FQ resistance was widespread amongst E coli isolates from poultry in Europe, with 53% of Spanish isolates and 30% of Dutch isolates demonstrating resistance. CAMPYLOBACTER JEJUNI No significant AMR was observed in the Australian retail food survey. No FQ resistance was detected and only 3.3% of isolates were resistant to erythromycin, 1.7% resistant to tetracycline, and 1.7% resistant to clindamycin. Almost all surveys conducted outside Australia reveal a situation with high levels of AMR, including widespread resistance to the FQ class of antimicrobial agent in Europe, but not yet emerged in Canada and the USA, though significant levels of resistance to nalidixic acid have been observed in the USA and this resistance is commonly a precursor of clinical FQ resistance. FQ resistance in the survey by de Jong et al (2009) was present in 60, 83, 48, 41 and 11% of C jejuni isolates from broiler caeca in Germany, Spain, France, Netherlands and UK respectively. ENTEROCOCCUS spp The enterococci are well known for accumulating resistance to a variety of antimicrobial agents. Reassuringly, the Australian retail food survey did not detect evidence of resistance to high level gentamicin or vancomycin (known as VRE), unlike the situation in Europe where VRE are still isolated, though at low levels (less than 6% in E faecium) Overall, isolates of commensal and zoonotic bacteria from poultry in Australia do not display significant antimicrobial resistance profiles, unlike the situation in most other countries. Antimicrobial Resistance in Australian Broilers: Summary Page 9 of 57

KEY FINDINGS FROM ANTIMICROBIAL RESISTANCE SURVEILLANCE REPORTS PERCENTAGE RESISTANCE IN BACTERIAL ISOLATES FROM BROILERS SALMONELLA SPECIES ANTIMICROBIAL AGENT Salmonella AUSTRALIA FSA 2008 Composite serovars CANADA CIPARS 2005 Composite 3 Provinces (range) FINLAND FINRES 2006 No poultry specific data DENMARK DANMAP 2007 Typhimurium EUROPE EFSA 2007 Typhimurium 7 MS + 1 HOLLAND MARAN 2005 Composite serovars USA NARMS 2006 Composite serovars NORWAY NORM 2006 <10 poultry isolates SWEDEN SVARM 2007 No poultry specific data Amikacin - 0 - - -- 0.0 Amoxy-clav 1 0-15.4 - - -- 19.1 Ampicillin, 4 4.8-23.1 10.0 18.6 60.0 22.4 6 amoxicillin Cefoxitin 1 0-15.4 - - - 18.4 Ceftiofur 0 0-15.4 0 - - 19.1 Chloramphenicol 0 0-4.8 10.0 13.6 2.0 2.6 0 Ciprofloxacin 0 0 0 0.9 34.0 0.0 0 Gentamicin 0 0-4.8 0 1.8 0 9.2 0 Kanamycin - 0-4.8 - - - 9.9 Nalidixic acid 1 0 0 12.3 30.0 0.7 61 Neomycin - - 0 0-0 Streptomycin 5 7.7-42.9 10.0 26.4-36.2 Sulfonamide - 0-14.3 10.0 21.4 50.0 23.0 Tetracycline 16 7.7-52.4 20.0 27.3 12.0 46.7 0 TMS 3 0 - - 1.3 Trimethoprim - - - 1.8 62.0 - Notes: Bold entries represent isolates obtained from food, normal font represents isolates from gut. TMS trimethoprim + sulphonamide. MS Member State SPAIN VAV 2005 Composite serovars

E COLI SPECIES ANTIMICROBIAL AGENT E coli AUSTRALIA FSA 2008 CANADA CIPARS 2005 3 Provinces (range) DENMARK DANMAP 2007 FINLAND FINRES 2006 EUROPE EFSA 2007 95% CI 12 MS + 2 HOLLAND MARAN 2005 USA NARMS 2006 NORWAY NORM 2006 SWEDEN SVARM 2007 Amikacin - 0 - - - - 0.0 -- - - Amoxy-clav 1 13.5-33.1 - - - - 11.5 - - Ampicillin, 38 24.7-49.3 10.5 12.3-19.9 40.8 53.9 20.1 7.6 5 64 amoxicillin Cefoxitin 0 9.9-23.4 - - - - 11.2 - - - Ceftiofur 0 3.7-24.6 1.8 0.0-1.0 - - 8.6 0.0 1 - Chloramphenicol 0 2.8-5.6 0 0.0-1.5 9.6 7.8 2.6 0.0 <1 19 Ciprofloxacin 0 0 14.0 0.3-2.7 17.4 29.6 0.0 0.0 7 53 Gentamicin 4 6.2-10.6 0 0.0-1.0 3.4 4.3 37.3 1.7 <1 11 Kanamycin 8 3.7-8.3 - - - - 11.5 0.0 2 - Nalidixic acid 0 0.7-4.9 10.5 0.3-2.7 37.8 33.0 5.0 0.0 7 89 Neomycin - - 0 3.5-8.3-12.2 - - - 16 Streptomycin 19 25.5-35.9 7.0 4.7-10.2 21.4-48.1 7.6 4 - Sulfonamide - 13.6-40.1 17.5 9.7-16.7 29.6 49.6 46.9 6.7 6 - Tetracycline 47 43.0-48.1 7.9 43.9 47.0 50.7 5.0 3 68 TMS 22 1.2-9.9 - - - 8.9 - - - Trimethoprim - 3.2-8.0 14.2 42.6-0.8 <1 - Notes: Bold entries represent isolates obtained from food, normal font represents isolates from gut. TMS trimethoprim + sulphonamide. MS Member State SPAIN VAV 2005 Antimicrobial Resistance in Australian Broilers: Summary Page 11 of 57

CAMPYLOBACTER JEJUNI SPECIES ANTIMICROBIAL AGENT Campylobacter jejuni AUSTRALIA FSA 2008 CANADA CIPARS 2005 3 Provinces (range) DENMARK DANMAP 2007 Domestic isolates (not imports) FINLAND FINRES 2006 EUROPE EFSA 2007 HOLLAND MARAN 2005 USA NARMS 2006 NORWAY NORM 2006 95% CI 9 MS + 2 cloacal samples SWEDEN SVARM 2007 Not surveyed in 2007 Ampicillin, - - - 0.0-6.9-44.3-6.5 44 amoxicillin Azithromycin - 0-14 - - - - 0.9 - Chloramphenicol 0 - - - 0-0 Ciprofloxacin 0 0 11.4 0.0-6.9 31.6 47.1 16.7 1.9 94 Cllindamycin 1.7 0-12.8 - - - - 0.7 - Erythromycin 3.3 0-14 1.8 0.0-6.9 3.7 2.9 0.9 0.0 6 Gentamicin 0 0 0 0.0-6.9 3.3 0 0 0.0 6 Nalidixic acid 0 0 11.4 0.0-6.9-47.1 16.7 1.9 93 Streptomycin - - 3.5-1.8 1.4 - - Tetracycline 1.7 59.0-69.8 9.6 0.5-11.4 29.6 50.0 47.2 0.0 44 Notes: Bold entries represent isolates obtained from food, normal font represents isolates from gut. MS Member State SPAIN VAV 2005 Antimicrobial Resistance in Australian Broilers: Summary Page 12 of 57

ENTEROCOCCUS FAECIUM SPECIES ANTIMICROBIAL AGENT Enterococcus faecium Ampicillin, AUSTRALIA FSA 2008 E faecium not isolated CANADA CIPARS 2005 3 Provinces (range) DENMARK DANMAP 2007 FINLAND FINRES 2006 95% CI EUROPE EFSA 2007 HOLLAND MARAN 2005 USA NARMS 2006 NORWAY NORM 2006 SWEDEN SVARM 2007 16.7-66.7 6.2 0.0-4.3 7.5 22.2 0.0 1 6 amoxicillin Avilamycin - 3.1 0.5-8.0 - - - - 83 Bacitracin 90.9-100 - 21.4-39.3 73.6-37.1 23 89 Chloramphenicol 0 0 0.0-4.3 0 0 0.0 0 8 Ciprofloxacin 0-16.7 - - 7.5 37.5 - - 50 Daptomycin 0 0 - - 1.6 - - - Erythromycin 50.0-66.7 29.7 6.8-20.0 34.0 9.6 21.3 11 47 Flavomycin (IR) 16.7-50.0 - - - 76.6 - - - Gentamicin 0 0 0.0-5.8 0 6.0 0.0 0 0 Kanamycin 0-16.7 3.1 - - 6.3 1.1 0 - Lincomycin - - - - 74.9 - - 86 Linezolid 0 0-3.8 0 0.0 0 - Narasin - - 77.8-91.8 - - 68.5 89 - Neomycin - - 0.0-4.3 - - - - - Nitrofurantoin 54.5-83.3 - - - 38.4 - - - QD 66.7-100 1.6-54.7 36.6 - - 61 Salinomycin - 75.0-24.5 - - - - Streptomycin 0-16.7 12.5 0.0-5.8 9.6 3.8 0.0 1 14 Tetracycline 81.8-100 10.9 19.0-36.4 52.8 53.0 11.2 16 75 Tylosin 50.0-63.6 - - - 7.9 - - - Vancomycin 0 1.6 0.7-8.5 5.7 0.0 1.1 0 0 Virginiamycin - - 3.5-14.5 - - 3.4 4 - Notes: Bold entries represent isolates obtained from food, normal font represents isolates from gut. MS Member State SPAIN VAV 2005 Antimicrobial Resistance in Australian Broilers: Summary Page 13 of 57

ENTEROCOCCUS FAECALIS SPECIES ANTIMICROBIAL AGENT Enterococcus faecalis Ampicillin, AUSTRALIA FSA 2008 CANADA CIPARS 2005 DENMARK DANMAP 2007 FINLAND FINRES 2006 95% CI EUROPE EFSA 2007 HOLLAND MARAN 2005 USA NARMS 2006 NORWAY NORM 2006 0 0 0.0-2.0 0.6 0.0 0.0 0 amoxycillin Avilamycin 0 0.0-2.6 - - - - Bacitracin 26.8-39.0 34.7-0.0 11 Chloramphenicol 1 0 0.0-2.6 3.5 0.0 7.1 0 Ciprofloxacin - - 4 0.8 - - Daptomycin 0 0 - - 0.0 - - Erythromycin 48 22.8 16.8-27.7 42.2 34.9 14.3 29 Flavomycin 43 1.8 4.0-10.9 9.2 0.0 - - Florfenicol 0 - - - - - Gentamicin 0 0 0.0-2.0 5.8 23.0 0.0 0 Kanamycin 9 0 - - 30.2 0.0 4 Linezolid 0 0-0 0.0 0.0 0 Narasin 10.2-19.4 - - 7.1 36 Neomycin 0.0-2.0 - - - - Salinomycin 3.5-5.2 - - - Streptomycin 5 3.5 1.0-5.6 24.9 10.3 7.1 0 Tetracycline 76 40.4 33.4-47.1 76.3 70.6 14.3 57 Tigecycline 6 0 - - 0.0 - - Tylosin 36.5 - - Vancomycin 0 0 0.0-2.0 1.2 0.0 0.0 0 SWEDEN SVARM 2007 SPAIN VAV 2005 Sulfonamide includes sulfisoxazole, sulfamethoxazole IR intrinsic resistance Bold numbers represent resistance % in retail meat samples MS Member State Antimicrobial Resistance in Australian Broilers: Summary Page 14 of 57

SUMMARY OF POULTRY CAMPYLOBACTER RESISTANCE LITERATURE KEY POINTS o Fluoroquinolone resistance not reported in Campylobacter isolates from Australian poultry o Recent surveillance has found macrolide resistance present at very low levels in Australian poultry isolates ABSTRACT Resistance by Campylobacter jejuni and C coli to the fluoroquinolone (FQ) class of antimicrobial agent is widespread throughout the world, in some countries reaching a prevalence of 80% amongst isolates from poultry. Sensitivity of campylobacer isolates to the fluoroquinolones is now a rare phenomenon throughout the world but present in Australia and in very few other countries. Macrolide resistance in C jejuni and C coli is also widespread globally. However, the prevalence of macrolide resistance is much lower than that encountered with the FQs. BACKGROUND FSANZ (2005) noted that Campylobacter is a leading cause of sporadic food-borne illness in Australia and that symptoms of the disease generally consist of self-limiting gastroenteritis, with hospitalisation required in a number of cases. Since the 1990s scientific attention has been focused on resistance of C. jejuni and C. coli to the fluoroquinolones (Smith et al, 2000). In Australia, fluoroquinolones have never been registered for use in food production animals. A case-control study for Campylobacter infection conducted in New South Wales from 1999 to 2001 (Unicomb et al., 2003), suggested that fluoroquinolone resistant Campylobacter are absent from Australia. Together with two laboratory-based surveys on antimicrobial resistance conducted in Western Australia (1999-2000) and the Australian Capital Territory (2001-2002), the case-control study found no resistance to fluoroquinolone in Campylobacter isolates known to be locally acquired from campylobacteriosis patients. Of the 370 Australian human Campylobacter isolates, 12 were resistant to fluoroquinolone, 10 of which were acquired from overseas travel. An Australian study of 79 Campylobacter isolates from chicken, found widespread resistance to erythromycin and significant resistance to doxycycline but no resistance to the fluoroquinolone enrofloxacin (Korolik et al., 1996). Another Australian study (Barton et al., 2001) of antimicrobial resistance of Campylobacter isolated from chickens reported significant resistance to ampicillin, ceftazidime and tetracycline in C. jejuni and C. coli isolates. No fluoroquinolone resistance was detected and there was relatively little resistance to the macrolides erythromycin or tylosin. As highlighted by Taylor and Courvalin (1988), all C. jejuni and C. coli isolates are intrinsically resistant to a number of antibiotics, including bacitracin, novobiocin, rifampin, streptogramin B, trimethoprim, vancomycin, and usually cephalothin, and so the choice of antimicrobial agent in sensitivity testing is important. A review of the current literature revealed 140 reports from 40 countries describing antimicrobial resistance in Campylobacter of broilers. There were reports from 34 countries describing the presence of FQR, 12 of which had >50% of isolates with FQR and up to 88% isolates with FQR. Only 4 countries (including Australia) found all isolates susceptible to the FQs. 22 countries Antimicrobial Resistance in Australian Broilers: Summary Page 15 of 57

reported ML resistance, with 13 observing <10% of isolates with ML R. 3 countries reported no isolates with ML R. Other resistances reported included those to ampicillin, azithromycin, chloramphenicol, clindamycin, gentamicin, kanamycin, nalidixic acid, sulphonamides, streptomycin and tetracycline COMPARATIVE RESISTANCE FREQUENCIES Country Australialiadieland Austr- Can- Eston- France Iceland Swed- Thai- UK USA Reference FSA 2009 Miflin et al 2007 Larkin et al 2006 Roasto et al Bywater et al Thorsteindottir Bywater et al Boonmar et al Bywater et al Son et al 2007 (food) (farm) 2007 2004 et al 2008 2004 2007 2004 Antimicrobial n=100 n=125 n=332 n=131 n=213 n=349 n=16 n=69 n=200 n=188 Agent Ampicillin 19 11 76 0 0 22 - Ciprofloxacin 0 0 4 73 32 1* 0 77 6 30 Erythromycin 4 0 2 8 0 0 17 2 0 Nalidixic acid 0 20 4 32 1 0 77 6 30 Tetracycline 1 19 50 58 0.3 0 26 35 99 * Enrofloxacin used to test FQ resistance in Iceland Antimicrobial resistance in C. jejuni isolates obtained from broiler caecum [de Jong et al 2009] (% of isolates) Antimicrobial Agent Resistance breakpoint France n=21 Germany n=15 Netherlands n=17 Spain n=12 UK n=47 Total n=112 Ciprofloxacin 4 mg/l 47.6 60.0 41.2 83.3 10.6 36.6 Erythromycin 32 mg/l 0 0 0 0 0 0 Gentamicin 16 mg/l 0 0 0 0 0 0 Nalidixic acid 32 mg/l 52.4 60.0 47.1 91.7 12.8 40.2 Tetracycline 16 mg/l 57.1 66.7 23.5 58.3 41.3 46.8 SUMMARY OF COUNTRY REPORTS OF CAMPYLOBACTER ANTIMICROBIAL RESISTANCE Country # Campy FQR Campy ML R Other R NOTES Refs AUSTRALIA 4 Enro 0% Ery 0-31% (coli 22%) Human isolates FQR, esp in travellers AUSTRIA 3 Cipro 62% Ery R 14.2% amp, tet, lin, jejuni, coli NA BELGIUM 1 Cipro 62% (coli) BRAZIL 2 Cipro (coli) Ery 80% amp, pen, sul, tet CANADA 5 Cipro <4% Ery < 7% tet, amp, clin, gen, strep CZECH REPUBLIC 5 Cipro 69% (coli 73%) Ery 1-14% NA, amp, tet, gen DENMARK 5 Enro 0-100% (farm to farm variation) Ery rare ESTONIA 3 Cipro 44-73% Ery 17% amp, NA, tet ETHIOPIA 1 Cipro 0% Ery <10% EUROPE 2 FQR to 80% in Sthn Europe FINLAND 1 Cipro Campylobacter spp. were isolated from 100% of organic broiler flocks, from 36.7% of conventional broiler flocks Antimicrobial Resistance in Australian Broilers: Summary Page 16 of 57

Country # Campy FQR Campy ML R Other R NOTES Refs FRANCE 6 Cipro 9-32% Ery 0-5% amp, tet GERMANY 5 Cipro 45% Ery 0.4% (coli 8%) GRANADA 1 Cipro 9% tet, amp HONG KONG 1 Cipro 85% ICELAND 1 Enro 3% IRAN 1 Cipro 69.4% Ery 0% amox, chl, gen, strep, tet IRELAND 4 Cipro 31.8% Ery 4.4% amp, tet, strep ITALY 1 FQR 18% (coli 54%) Ery 3% (coli 23%) JAPAN 12 Cipro 12% Ery 0% (coli 48%) KOREA 3 Cipro 87.9% Ery 19.4 amp, chl, tet, kan, NA MALAYSIA 1 Ery 22-26% amp, chl, kan, tet NETHERLANDS 3 Oflox 29% NORTHERN 2 Cipro 3-7% Ery <14% IRELAND NORWAY 2 FQR 1.2% amp, oxy POLAND 2 Cipro 55.9% Ery 0% amp, tet PORTUGAL 3 Cipro 80.5% Ery 5.1% amp, tet PUERTO RICO 1 Cipro 0% Ery few R RUSSIA 1 FQR 0% Ery <1% SENEGAL 1 FQR 43.4% SLOVENIA 1 Cipro 11% Ery 11% SPAIN 3 Cipro 72% Ery low% SWEDEN 4 FQR 2% SWITZERLAND 4 Cipro 28.7% amp, tet, sul TAIWAN 1 FQR THAILAND 8 Cipro 58.8-77% amp, sul, tet TRINIDAD 1 Cipro 86.6% TURKEY 1 FQR 31.3% UNITED 13 Cipro 11-17% Ery 5% Imported vs domestic KINGDOM USA 21 Cipro 14-46% Ery 2% azi, chl, clin, gen, NA, tet Antimicrobial Resistance in Australian Broilers: Summary Page 17 of 57

SUMMARY OF POULTRY SALMONELLA RESISTANCE LITERATURE ABSTRACT KEY POINTS o Absence of endemic Salmonella Enteritidis in broiler chickens o Absence of multiresistant Salmonella Typhimurium DT104 o Fluoroquinolone resistance not reported in Salmonella isolates from Australian poultry o ESBL resistance not reported in Salmonella isolates from Australian poultry Salmonellosis is one of the most frequently reported food-borne illnesses in Australia, leading to significant morbidity in the population with the majority of cases experiencing symptoms limited to gastroenteritis. Salmonella Typhimurium FSANZ (2005) noted that of recent concern worldwide is the emergence of multiple antibiotic resistant strains of Salmonella, an example being S. Typhimurium definitive phage type 104 (DT104). Multi-resistant S. Typhimurium DT104 is a significant human and animal pathogen, with high morbidity observed in cattle and poultry (Crerar et al., 1999). To date, this organism is not endemic in Australia, although it is a significant health problem in European countries, North America, the Middle East, South Africa and South-East Asia (Jay et al., 2003). S. Typhimurium DT104 constitutes 8 9% of human Salmonella isolates in the USA. Sporadic human cases are reported in Australia, although these are commonly acquired overseas (Blumer et al., 2003). During 2001 an outbreak of S. Typhimurium DT104 occurred in Victoria and was linked to consumption of contaminated imported halva (a sesame seed product). In a review of food safety in Australia, Abelson et al 2006 describe only one instance of multi-drug resistant salmonellosis, being this same incident of S. Typhimurium 104 contaminating halva from Turkey. The investigation of the S. Typhimurium 104 outbreak indicated a high rate of hospitalisation with 33% (8/24) of notified cases requiring admission. In Australia, this outbreak investigation prevented an estimated 87.3% of the total consignment of halva reaching consumers. This investigation may have prevented 79 notified cases and a further 1,185 infections in the community, saving as much as $1.3 million. Salmonella Enteritidis Epidemiological investigations in Europe and the USA have identified that raw or undercooked egg and egg products were the foods most commonly associated with increased illness in humans due to S. Enteritidis. However, consumption of chicken meat has recently been implicated as a source of sporadic S. Enteritidis infection in the US. Fortunately S. Enteritidis (in particular phage type 4; PT 4) is not endemic in Australia, with most human cases reported from travellers returning from overseas. Queensland appears to have the greatest number of locally acquired infections, with phage type 26 being the most common. Antimicrobial Resistance in Australian Broilers: Summary Page 18 of 57

Annual Report* Salmonella Enteritidis AUSTRALIA Number Salmonella Enteritidis isolates raw chicken meat, chicken processing environment** 2008 4/4134 0.10% 2007 1/3842 0.03% 2006 15/4386 0.34% 2005 27/6011 0.45% Egg and egg products Human Comments 0 179/1810 Chicken meat isolate from QLD. Phage types 26 (3) & RDNC (1) 1/102 147/2185 Chicken meat isolate from QLD. Phage type 26 0 124/1792 Chicken meat isolates from QLD. Phage types 26 (10), 35 (1), RDNC (1), untypable (3) 0/84 151/1713 Chicken meat isolates from QLD. Phage types 14 (14), 14var (6), 14b (1), 26 (4), untypable (2) 2004 0/2787 1/65 168/1551 No chicken meat isolates 2003 0/1866 0/3 123/1437 No chicken meat isolates 2002 0/2204 0/2 156/1796 No chicken meat isolates 2001 0/2190 0/11 114/1772 No chicken meat isolates 2000 0/1925 0/1 109/1453 No chicken meat isolates 1999 0/1696 0/0 176/2193 No chicken meat isolates *Australian Salmonella Reference Centre ** Of 18,373 broiler isolates from 2005 to 2008, only 47 Enteritidis have been identified and all originated in Queensland, 18 of which were phage type 26 and none the epidemic phage type 4 which is a significant food safety concern outside Australia, especially in eggs and egg products. Salmonella Sofia Based on industry data and retail survey studies, S. Sofia is the most commonly isolated Salmonella serovar from chickens in Australia. In common with other Salmonella enterica subsp. salamae serovars (previously described as subspecies II), S. Sofia is regarded as having relatively low virulence to humans. This is confirmed by epidemiological data from the National Enteric Pathogens Surveillance Scheme, where S. Sofia is infrequently isolated from humans. Challenge studies to determine if S. Sofia could act to competitively exclude more pathogenic Salmonella serovars concluded that S. Sofia does not exclude the virulent serovar S. Typhimurium as both serovars could co-colonise regardless of initial colonisation status. In surveys of antimicrobial resistance of Salmonella isolates in Australia, most streptomycin resistance was detected in the non-pathogenic serovar subsp II Sofia. For example, in 2006 87.3% of 534 Sofia isolates tested were resistant to streptomycin, while only 6.2% of 1084 non- Sofia broiler chicken isolates were resistant. Antimicrobial Resistance in Australian Broilers: Summary Page 19 of 57

Antimicrobial Resistance in Salmonella isolates In Australia, antimicrobial susceptibility testing of Salmonella isolates of both human and non-human origin submitted to the Australian Salmonella Reference Centre (ASRC) commenced in May 2000. The following table summarises the results of susceptibility testing of 10,248 salmonella isolates from Australian broilers submitted to the ASRC and reported in annual reports between 2001 and 2008. The resistance of isolates to a panel of 11 antimicrobial agents is summarised below. Resistance to all agents is low or absent and multiple resistance is also present at a low frequency. There is also a trend for reduced levels of resistance in the time period from 2001 to 2008. Highest resistance levels are reported for the aminoglycoside, streptomycin. This apparently high frequency appears to be due to the high prevalence of this resistance in the avirulent S Sophia, the dominant Salmonella serovar isolated from broilers. Antimicrobial susceptibility testing of Salmonella isolates from broilers submitted to the Australian Salmonella Reference Centre Year Strains tested Gen 4μg/ml Kan 16μg/ml Nal 16μg/ml Chl 8μg/ml Amp 8μg/ml Tet 8μg/ml Str 1 16μg/ml Sul 256μg/ml Tmp 4μg/ml Cip 1&4μg/ml Cef 1μg/ml Multiple Resistance 2 2008 1408 0 2.2 0 0.2 5.8 12.4 37.9 7.7 6.0 0 0 3.7 2007 1364 0 1.8 0.1 0.1 5.1 11.1 32.6 7.4 6.2 0 0.1 2.6 2006 1618 0.1 2.1 0.2 0.4 5.0 13.0 32.9 6.2 5.9 0 0 3.5 2005 1925 0 3.0 0.2 0.9 6.2 10.3 47.0 7.6 7.3 0 0 4.5 2004 1008 0.3 3.6 0 1.5 6.5 9.3 45.0 5.8 6.7 0 0 2.9 2003 963 0 1.3 0.1 0.7 9.6 22.5 31.8 4.8 5.1 0 0 2.7 2002 886 0 1.1 0 1.4 6.8 16.8 46.8 6.9 8.0 0 0 3.2 2001 1076 0 1.1 0.5 3 1.0 9.9 18.2 3 74.3 3 16.4 15.2 0 NR NR Gen gentamicin; Kan kanamycin; Nal nalidixic acid; Chl chloramphenicol; Amp ampicillin; Tet tetracycline; Str streptomycin; Sul sulfadiazine; Tmp trimethoprim; Cip ciprofloxacin; Cef cefotaxime (3GC) 1 Most streptomycin resistance was detected in the non-pathogenic serovar subsp II Sofia. For example, in 2006 87.3% of 534 Sofia isolates tested were resistant to streptomycin, while only 6.2% of 1084 non-sofia broiler chicken isolates were resistant. In 2008, S II Sofia constituted 45.9% of the broiler isolates. Streptomycin resistance was 91.6% and 3.9% amongst Sofia other serovars respectively. 2 Multiple resistance defined as Salmonella resistant to 4 or more antibiotics [2008 comparison of salmonella isolates from broiler 3.7%; bovine 10.6; porcine 30.4] 3 Antibiotic concentrations lower than other years. Nal 8μg/ml; Tet 4μg/ml and Str 4μg/ml Of 9,172 salmonella isolates from broilers assessed for cefotaxime resistance between 2002 and 2008 inclusive, only one isolate (serovar Saintpaul isolated in 2007) displayed phenotypic resistance. Of 10,248 salmonella isolates from broilers assessed for ciprofloxacin resistance between 2001 and 2008 inclusive, no isolates have displayed resistance. Antimicrobial Resistance in Australian Broilers: Summary Page 20 of 57

The emergence and dissemination of extra-spectrum beta-lactamase (ESBL) resistance in Salmonella isolates from a variety of animal species including poultry has the potential to compromise the treatment of humans with serious infections with zoonotic serotypes of Salmonella. While ESBL resistance has not been identified in Australia, the following tables demonstrate its widespread occurrence in Europe, North America, Africa and Asia. Salmonella ESBL producers in animals and food of animal origin β-lactamase Organism No strains Country Source Reference SHV-12 S 35:c:1,2 1 Senegal Poultry Cardinale et al 2001 TEM-52 S Agona 3 Belgium Poultry Cloeckaert et al 2007 TEM-52 S Blockley 6 Netherlands Poultry Hasman et al 2005 TEM-52 S Derby 1 Belgium Poultry Cloeckaert et al 2007 CTX-M-9 S Enteriditis 1 Spain Laying hen Riano et al 2006 TEM-52 S Infantis 1 Belgium Poultry Cloeckaert et al 2007 TEM-52 S Paratyphi B 1 Netherlands Poultry Hasman et al 2005 SHV-12 S Typhimurium 1 Netherlands Poultry Hasman et al 2005 TEM-52 S. Typhimurium 3 Belgium Poultry Jensen et al 2006 CTX-M-2 S. Virchow 1 Netherlands Broiler Hasman et al 2005 CTX-M-2 S. Virchow 1 Ireland Poultry Hopkins et al 2006 CTX-M-2 S. Virchow 6 Belgium Poultry Bertrand et al 2006 CTX-M-9 S. Virchow 8 France Poultry Weill et al 2004 CTX-M-9 S. Virchow 2 Spain Broiler Riano et al 2006 CTX-M-32 S. Virchow 2 Greece Poultry Politi et al 2005 Carattoli, A. (2008). "Animal reservoirs for extended spectrum beta-lactamase producers." Clin Microbiol Infect 14 Suppl 1: 117-23 A total of 229 publications were reviewed which described antimicrobial resistance in Salmonella isolates form 49 countries and one region. Ten countries reported multi-drug resistance (MDR) in S Enteritidis while 38 described MDR in other Salmonella serovars. Importantly extra-spectrum beta-lactamase resistance (EBSL) was identified in 15 countries and quinolone / fluoroquinolone resistance in 31 countries SUMMARY OF COUNTRY REPORTS OF SALMONELLA ANTIMICROBIAL RESISTANCE COUNTRY # pubs S Typhimurium MDR S enteritidis MDR AUSTRALIA 4 low level MDR NOT REPORTED AUSTRIA 1 MDR BANGLADE 2 MDR SH BELGIUM 6 typical SGI1 MDR profile (Ap Cm Ff Sm Sp Su Tc) Salmonella ESBL NOT REPORTED blactx-m Salmonella FQR NOT REPORTED FQR (reduced sensitivity) Comments BRAZIL 5 MDR MDR NAR 86% 100% of the isolated Salmonella strains had resistance to ampicillin and tetracycline Antimicrobial Resistance in Australian Broilers: Summary Page 21 of 57

COUNTRY # pubs S Typhimurium MDR S enteritidis MDR Salmonella ESBL Salmonella FQR Comments BULGARIA 1 MDR MDR NAR 20% most common resistance patterns were to ampicillin, carbenicillin, tetracycline and erythromycin simultaneously with or without resistance to nalidixic acid CANADA 13 MDR MDR ESBL NAR 96% RSC (reduced sensitivity to cipro) AMR to ampicillin, chloramphenicol, kanamycin, neomycin, streptomycin, sulfisoxazole, and tetracycline DT104 and pentaresistance ACSSuT CHILE 1 NAR 41% CHINA, PEOPLE S REPUBLIC OF 1 MDR ESBL COLUMBIA 1 low level AMR low level AMR CROATIA 1 MDR FQR DENMARK 9 FQR FQR some patients in Denmark acquired S. Corvallis from imported chicken meat from Thailand multiple resistance was most often acquired outside Denmark EGYPT 1 MDR ETHIOPIA 3 MDR EUROPE 1 MDR DT104 and pentaresistance ACSSuT FINLAND 1 MDR Antimicrobial Resistance in Australian Broilers: Summary Page 22 of 57

COUNTRY # pubs S Typhimurium MDR FRANCE 7 MDR ACSSpSuTe S enteritidis MDR GERMANY 9 MDR FQR NAR Salmonella ESBL Salmonella FQR Comments ESBL NAR spectacular increase of the resistance frequency to nalidixic acid for the strains belonging to serotypes Hadar and Virchow, especially in the poultry food sector (14% in 1993 vs. 72% in 2000 for Salmonella Virchow, 4% in 1993 vs. 70% in 2000 for Salmonella Hadar) Salmonella serovar Typhimurium DT104 qnrs gene GREECE 2 ESBL HUNGARY 1 MDR NAR ICELAND 1 MDR FQR INDIA 3 MDR FQR IRAN 2 MDR ESBL NAR IRELAND 1 MDR ISRAEL 1 NAR ITALY 3 MDR DT104 JAPAN 13 MDR DT104 KOREA 5 MDR MDR ESBL NAR KUWAIT 1 MDR FQR LITHUANIA 1 ESBL FQR MALAYSIA 3 MDR MEXICO 4 blacmy-2 NAR NETHERLA NDS 7 ESBL NAR qnr genes NEW 1 MDR ZEALAND NIGERIA 1 MDR FQR OMAN 1 MDR PHILIPPINE 1 MDR S POLAND 5 MDR NAR DT104 PORTUGAL 1 FQR 50% SENEGAL 2 MDR MDR NAR SERBIA 1 MDR SOUTH 2 MDR MDR AFRICA SPAIN 8 MDR MDR ESBL NAR DT104 TAIWAN 1 FQR Antimicrobial Resistance in Australian Broilers: Summary Page 23 of 57

COUNTRY # pubs S Typhimurium MDR S enteritidis MDR Salmonella ESBL Salmonella FQR Comments THAILAND 12 MDR ESBL FQR Seven of 14 isolates from persons in Denmark had patterns found in persons and chicken meat in Thailand TURKEY 2 MDR ESBL FQR UNITED KINGDOM UNITED STATES OF AMERICA 32 MDR MDR ESBL FQR frozen chicken portions (n = 150) imported from Brazil and Thailand (generally for manufacturing and catering) were also tested. Brazilian salmonellas showed no multiple resistance, but an isolate of Salmonella Virchow from Thai chickens was resistant to two antibiotics. 38 MDR MDR Ceftiofur <30% CMY-2 FQR VENEZUELA 2 MDR FQR VIETNAM 4 MDR FQR enro 22% DT104 + flost nine Salmonella isolates (14.52%) were multiresistant to at least nine antibiotics Antimicrobial Resistance in Australian Broilers: Summary Page 24 of 57

SUMMARY OF POULTRY ESCHERICHIA COLI RESISTANCE LITERATURE KEY POINTS o No reports of fluoroquinolone (FQ) resistance amongst E coli isolates from poultry in Australia o No reports of Extended Spectrum Beta Lactamase (ESBL) resistance amongst E coli isolates from poultry in Australia BACKGROUND ABSTRACT Escherichia coli (E coli) is the prototypic commensal species of the facultative anaerobic colonic microflora of most animal species, including mammals and birds. Over the last half-century it has become increasingly obvious that there are a number of different enteropathogenic groups of E. coli. At least six known pathotypes associated with gastrointestinal infections have been recognized, with other pathotypes causing extraintestinal infections such as urinary tract infections, septicaemia, and meningitis in humans and a number of similar diseases in animals. Being typical E. coli in all respects, apart from possessing their respective virulence factors, these pathogenic E. coli behave in all respects including biochemically and ecologically like other nonpathogenic E. coli. Thus, if no distinct markers are available, their detection among commensal E. coli becomes very difficult The pathotypes associated with gastrointestinal infections currently recognized include: o Enteropathogenic E. coli (EPEC) o Enterotoxigenic E. coli (ETEC) o Verocytotoxigenic E. coli (VTEC) or Shiga toxin-producing E coli (STEC) of which Enterohaemorrhagic E. coli (EHEC) (a human pathogenic subgroup of VTEC that causes haemorrhagic colitis (HC) and the haemolytic uraemic syndrome (HUS)) are a subgroup. o Enteroinvasive E. coli (EIEC) o Enteroaggregative E. coli (EAggEC or EAEC) o Diffusely-adherent E. coli (DAEC). The Shiga toxin-producing E. coli O157:17 is well described in cattle, goat, sheep, and other farmed animals (where it can cause disease but is commonly an asymptomatic coloniser of the lower gut), with cattle being the primary source of this pathogen. STEC are very uncommonly isolated from poultry. Pathogenic E. coli are also important causes of extraintestinal infections in humans and include the following pathotypes: o Extraintestinal pathogenic Escherichia coli (ExPEC) o uropathogenic E. coli (UPEC) o sepsis-associated E. coli (SEPEC) o neonatal meningitis-associated E. coli (NEMEC) An additional animal pathotype (avian pathogenic E. coli (APEC)) causes extraintestinal infections (primarily respiratory infections, pericarditis, and septicaemia) in poultry, but does not seem to have a close counterpart in human disease, although an autotransporter hemagglutinin believed to Antimicrobial Resistance in Australian Broilers: Summary Page 25 of 57