Antimicrobial Resistance Profiles and Diversity in Salmonella from Humans and Cattle, 2004–2011

Analysis of long‐term anti‐microbial resistance (AMR) data is useful to understand source and transmission dynamics of AMR. We analysed 5124 human clinical isolates from Washington State Department of Health, 391 cattle clinical isolates from the Washington Animal Disease Diagnostic Laboratory and 1864 non‐clinical isolates from foodborne disease research on dairies in the Pacific Northwest. Isolates were assigned profiles based on phenotypic resistance to 11 anti‐microbials belonging to eight classes. Salmonella Typhimurium (ST), Salmonella Newport (SN) and Salmonella Montevideo (SM) were the most common serovars in both humans and cattle. Multinomial logistic regression showed ST and SN from cattle had greater probability of resistance to multiple classes of anti‐microbials than ST and SN from humans (P < 0.0001). While these findings could be consistent with the belief that cattle are a source of resistant ST and SN for people, occurrence of profiles unique to cattle and not observed in temporally related human isolates indicates these profiles are circulating in cattle only. We used various measures to assess AMR diversity, conditional on the weighting of rare versus abundant profiles. AMR profile richness was greater in the common serovars from humans, although both source data sets were dominated by relatively few profiles. The greater profile richness in human Salmonella may be due to greater diversity of sources entering the human population compared to cattle or due to continuous evolution in the human environment. Also, AMR diversity was greater in clinical compared to non‐clinical cattle Salmonella, and this could be due to anti‐microbial selection pressure in diseased cattle that received treatment. The use of bootstrapping techniques showed that although there were shared profiles between humans and cattle, the expected and observed number of profiles was different, suggesting Salmonella and associated resistance from humans and cattle may not be wholly derived from a common population.     

Prevalence,Serovars and Antimicrobial Susceptibility of Salmonella spp. from Wild and Domestic Green Iguanas (Iguana iguana) in Grenada,West Indies

Cloacal swabs from 62 green iguanas (Iguana iguana), including 47 wild and 15 domestic ones from five parishes of Grenada, were sampled during a 4‐month period of January to April 2013 and examined by enrichment and selective culture for the presence of Salmonella spp. Fifty‐five per cent of the animals were positive, and eight serovars of Salmonella were isolated. The most common serovar was Rubislaw (58.8%), a serovar found recently in many cane toads in Grenada, followed by Oranienburg (14.7%), a serovar that has been causing serious human disease outbreaks in Japan. Serovar IV:48:g,z₅₁:‐ (formerly, S. Marina) highly invasive and known for serious infections in children in the United States, constituted 11.8% of the isolates, all of them being from domestic green iguanas. Salmonella Newport, a serovar recently found in a blue land crab in Grenada, comprised 11.8% of the isolates from the green iguanas. The remaining four less frequent serovars included S. Javiana and S. Glostrup. Antimicrobial susceptibility tests conducted by a disc diffusion method against amoxicillin–clavulanic acid, ampicillin, cefotaxime, ceftazidime, ciprofloxacin, enrofloxacin, gentamicin, nalidixic acid, streptomycin, tetracycline and trimethoprim–sulfamethoxazole showed that drug resistance is minimal, with intermediate susceptibility, mainly to streptomycin, tetracycline and cefotaxime. This is the first report of isolation and antimicrobial susceptibilities of various Salmonella serovars from wild and domestic green iguanas in Grenada, West Indies.     

Phenotypic and Genotypic Characterization of Salmonella enterica in Captive Wildlife and Exotic Animal Species in Ohio,USA

The purpose of this study was to investigate the occurrence, antimicrobial resistance patterns, phenotypic and genotypic relatedness of Salmonella enterica recovered from captive wildlife host species and in the environment in Ohio, USA. A total of 319 samples including faecal (n = 225), feed (n = 38) and environmental (n = 56) were collected from 32 different wild and exotic animal species in captivity and their environment in Ohio. Salmonellae were isolated using conventional culture methods and tested for antimicrobial susceptibility with the Kirby–Bauer disc diffusion method. Salmonella isolates were serotyped, and genotyping was performed using the pulsed‐field gel electrophoresis (PFGE). Salmonella was detected in 56 of 225 (24.9%) faecal samples; six of 56 (10.7%) environmental samples and six of 38 (15.8%) feed samples. Salmonella was more commonly isolated in faecal samples from giraffes (78.2%; 36/46), cranes (75%; 3/4) and raccoons (75%; 3/4). Salmonella enterica serotypes of known public health significance including S. Typhimurium (64.3%), S. Newport (32.1%) and S. Heidelberg (5.3%) were identified. While the majority of the Salmonella isolates were pan‐susceptible (88.2%; 60 of 68), multidrug‐resistant strains including penta‐resistant type, AmStTeKmGm (8.8%; six of 68) were detected. Genotypic diversity was found among S. Typhimurium isolates. The identification of clonally related Salmonella isolates from environment and faeces suggests that indirect transmission of Salmonella among hosts via environmental contamination is an important concern to workers, visitors and other wildlife. Results of this study show the diversity of Salmonella serovars and public health implications of human exposure from wildlife reservoirs.     

Prevalence of Salmonella spp. in Cane Toads (Bufo marinus) from Grenada,West Indies,and their Antimicrobial Susceptibility

Cloacal swabs and caecal contents sampled from 58 cane toads (Bufo marinus) in St George’s parish, Grenada, during a 7‐month period in 2011 were examined by an enrichment and selective culture method for presence of Salmonella spp. Twenty‐four (41%) toads were positive for Salmonella spp. of which eight were Salmonella enterica serovar Javiana, and eight were S. enterica serovar Rubislaw. The other serovars were as follows: Montevideo, 6; Arechavaleta, 1; and serovar: IV:43:‐:‐, 1. The high frequency of isolation of serovar Javiana, an emerging human pathogen associated with several outbreaks in the recent years in the eastern United States, suggests a possible role for cane toads in transmission of this serovar. Although S. Rubislaw has been isolated from lizards, bats and cases of some human infections, there is no report of its carriage by cane toads, and in such high frequency. The rate of carriage of S. Montevideo, a cause for human foodborne outbreaks around the world was also over 10% in the 58 toads sampled in this study. The antimicrobial drug susceptibility tests against amoxicillin‐clavulanic acid, ampicillin, cefotaxime, ceftazidime, ciprofloxacin, enrofloxacin, gentamicin, imipenem, nalidixic acid, streptomycin, tetracycline and trimethoprim‐sulfamethoxazole showed that drug resistance is minimal and is of little concern. Antimicrobial resistance was limited to ampicillin and amoxicillin‐clavulanic acid in one isolate of S. Javiana and one isolate of S. Rubislaw. This is the first report of isolation and antimicrobial susceptibilities of various Salmonella serovars not identified previously in cane toads in Grenada, West Indies.     

Salmonella enterica serovar Kentucky recovered from human clinical cases in Maryland,USA (2011–2015)

Salmonella Kentucky is among the most frequently isolated S. enterica serovars from food animals in the United States. Recent research on isolates recovered from these animals suggests there may be geographic and host specificity signatures associated with S. Kentucky strains. However, the sources and genomic features of human clinical S. Kentucky isolated in the United States remain poorly described. To investigate the characteristics of clinical S. Kentucky and the possible sources of these infections, the genomes of all S. Kentucky isolates recovered from human clinical cases in the State of Maryland between 2011 and 2015 (n = 12) were sequenced and compared to a database of 525 previously sequenced S. Kentucky genomes representing 12 sequence types (ST) collected from multiple sources on several continents. Of the 12 human clinical S. Kentucky isolates from Maryland, nine were ST198, two were ST152, and one was ST314. Forty‐one per cent of isolates were recovered from patients reporting recent international travel and 58% of isolates encoded genomic characteristics similar to those originating outside of the United States. Of the five isolates not associated with international travel, three encoded antibiotic resistance genes conferring resistance to tetracycline or aminoglycosides, while two others only encoded the cryptic aac(6′)‐Iaa gene. Five isolates recovered from individuals with international travel histories (ST198) and two for which travel was not recorded (ST198) encoded genes conferring resistance to between 4 and 7 classes of antibiotics. Seven ST198 genomes encoded the Salmonella Genomic Island 1 and substitutions in the gyrA and parC genes known to confer resistance to ciprofloxacin. Case report data on food consumption and travel were, for the most part, consistent with the inferred S. Kentucky phylogeny. Results of this study indicate that the majority of S. Kentucky infections in Maryland are caused by ST198 which may originate outside of North America.     

Genomic diversity and resistome profiles of Salmonella enterica subsp. enterica serovar Kentucky isolated from food and animal sources in Ireland

Salmonella enterica subsp. enterica serovar Kentucky is frequently isolated from poultry, dairy and beef cattle, the environment and people with clinical salmonellosis globally. However, the sources of this serovar and its diversity and antimicrobial resistance capacities remain poorly described in many regions. To further understand the genetic diversity and antimicrobial sensitivity patterns among S. Kentucky strains isolated from non-human sources in Ireland, we sequenced and analysed the genomes of 61 isolates collected from avian, bovine, canine, ovine, piscine, porcine, environmental and vegetation sources between 2000 and 2016. The majority of isolates (n = 57, 93%) were sequence type (ST) 314, while only three isolates were ST198 and one was ST152. Several isolates were multidrug-resistant (MDR) and 14 carried at least one acquired antimicrobial resistance gene. When compared to a database of publicly available ST314, four distinct clades were identified (clades I–IV), with the majority of isolates from Ireland clustering together in Clade I. Two of the three ST198 isolates were characteristic of those originating outside of the Americas (Clade ST198.2), while one was distantly clustered with isolates from South and North America (Clade ST198.1). The genomes of the two clade ST198.2 isolates encoded Salmonella Genomic Island 1 (SGI1), were multidrug-resistant and encoded polymorphisms in the DNA gyrase (gyrA) and DNA topoisomerase (parC) known to confer resistance to fluoroquinolones. The single ST152 isolate was from raw beef, clustered with isolates from food and bovine sources in North America and was pan-susceptible. Results of this study indicate that most S. Kentucky isolates from non-human sources in Ireland are closely related ST314 and only a few isolates are antimicrobial-resistant. This study also demonstrates the presence of multidrug-resistant ST198 in food sources in Ireland.     

Prevalence and characterization of multidrug resistance and variant Salmonella genomic island 1 in Salmonella isolates from cattle,poultry and humans in Iran

Salmonella enterica is a common food‐borne pathogen with occasional multidrug resistance (MDR). Salmonella genomic island (SGI1) is a horizontally transmissible genomic island, containing an MDR gene cluster. All Salmonella serotypes are public health concern, although there is an additional concern associated with those that harbour SGI1. In Iran, there are no data on the presence of SGI1 variants in Salmonella isolates. The present study was conducted to identify MDR‐ and SGI1‐carrying Salmonella strains isolated from various sources and to compare their genetic relatedness between human and animal sources. In total, 242 Salmonella isolates collected from chicken, cattle, and humans from 2008 through 2014 were studied. The isolates were tested for resistance to 14 antimicrobials via the disc diffusion method. They were also tested for the presence of SGI1 variants via PCR, and genetic relatedness was evaluated based on pulsed‐field gel electrophoresis (PFGE). Resistance to at least one antimicrobial agent was observed in 132 (54%) Salmonella isolates (n = 242), while more than 40% of the isolates showed MDR. Based on PCR analysis, eight variants of SGI1, including SGI1, SGI1‐B, SGI1‐C, SGI1‐D, SGI1‐F, SGI1‐I, SGI1‐J and SGI1‐O, were found in both human and animal isolates. Statistical analysis revealed no significant difference in the prevalence of SGI1 variants between human and animal isolates (p > 0.05). Macrorestriction PFGE analysis of the isolates with the same SGI1 variant and resistance patterns revealed genetic relatedness ranging from 70% to 100% among human and animal isolates. According to our review, this is the first documentation of SGI1 in Salmonella isolates in Iran. The presence of similar SGI1 variants in both humans and animals, along with their related PFGE patterns, suggests that food‐producing animals may be a source of MDR Salmonella isolates in Iran.     

Salmonella Serovars and Antimicrobial Resistance Profiles in Beef Cattle,Slaughterhouse Personnel and Slaughterhouse Environment in Ethiopia

The present study was undertaken to determine the occurrence, distribution and antimicrobial resistance profiles of Salmonella serovars in slaughter beef cattle, slaughterhouse environment and personnel engaged in flaying and evisceration during slaughtering process. A total of 800 samples (each sample type, n = 100) consisting of swabs from hides, slaughterhouse personnel hands at flaying and evisceration, rumen and caecal contents, mesenteric lymph nodes, carcasses and holding pens were collected. Of the total 100 beef cattle examined, 14% were Salmonella positive in caecal content and/or mesenteric lymph nodes. Of the various samples analysed, Salmonella was detected in 31% of hides, 19% of rumen contents, 8% of mesenteric lymph nodes, 6% of caecal contents, 2% of carcass swabs, 9% of palm swabs taken from the hands of personnel in the slaughterhouse during flaying (7%) and evisceration (2%), and in 12% of holding pen swabs. The Salmonella isolates (n = 87) belonged to eight different serovars of which S. Anatum (n = 54) and S. Newport (19) were the major serovars and both serovars were detected in all sample sources except in carcass swabs. Eighteen of the 87 (20.7%) Salmonella serovars consisting of Newport (n = 14), Anatum (n = 3) and Eastbourne (n = 1) were resistant to one or more antimicrobials. Among the antimicrobial resistant Salmonella serovars, S. Newport was multidrug resistant (15.6%) and exhibited resistance to streptomycin, sulphisoxazole and tetracycline.     

Comparative Phenotypic and Genotypic Analyses of Salmonella Rissen that Originated from Food Animals in Thailand and United States

Salmonella enterica serovar Rissen has been recognized as one of the most common serovar among humans and pork production systems in different parts of the world, especially Asia. In the United States, this serovar caused outbreaks but its epidemiologic significance remains unknown. The objectives of this study were to compare the phenotypic (antimicrobial susceptibility) and genotypic attributes of Salmonella Rissen isolated in Thailand (Thai) and the United States (US). All the Thai isolates (n = 30) were recovered from swine faecal samples. The US isolates (n = 35) were recovered from swine faecal samples (n = 29), cattle (n = 2), chicken (n = 2), dog (n = 1) and a ready‐to‐eat product (n = 1). The antimicrobial susceptibility of isolates was determined using the Kirby‐Bauer disk diffusion method with a panel of 12 antimicrobials. Pulse‐field gel electrophoresis (PFGE) was used to determine the genotypic diversity of isolates. All Thai isolates showed multidrug resistance (MDR) with the most frequent antibiotic resistance shown against ampicillin (100%), sulfisoxazole (96.7%), tetracycline (93.3%), streptomycin (90%) and chloramphenicol (30%). About half of the isolates of USA origin were pan‐susceptible and roughly 30% were resistant to only tetracycline (R‐type: Te). Salmonella Rissen isolated from Thailand and the USA in this study were found to be clonally unrelated. Genotypic analyses indicated that isolates were clustered primarily based on the geographic origin implying the limited clonality among the strains. Clonal relatedness among different host species within the same geography (USA) was found. We found genotypic similarity in Thai and US isolates in few instances but with no epidemiological link. Further studies to assess propensity for increased inter‐regional transmission and dissemination is warranted.     

Prevalence,Characterization and Antibiotic Resistance of Salmonella Isolates in Large Corvid Species of Europe and North America Between 2010 and 2013

It is well understood that Salmonella is carried by animals and in majority of cases as asymptomatic hosts. Surveillance efforts have focused on the role of agriculture and contamination points along the food chain as the main source of human infection; however, very little attention has been paid to the contribution of wildlife in the dissemination of Salmonella and what effect anthropogenic sources have on the circulation of antibiotic resistant Salmonella serovars in wildlife species. A purposive survey was taken of large corvids roosting yearly between November and March in Europe and North America. Two thousand and seven hundred and seventy‐eight corvid faecal specimens from 11 countries were submitted for Salmonella spp. culture testing. Presumptive positive isolates were further serotyped, susceptibility tested and analysed for antibiotic resistance genes. Overall, 1.40% (39/2778) (CI = 1.01, 1.90) of samples were positive for Salmonella spp. Salmonella Enteritidis was the most prevalent serovar followed by S. Infantis, S. Montevideo and S. Typhimurium. No significant difference (P > 0.05) was found in the proportion of Salmonella recovered in Europe versus North America. The most variability of serovars within a site was in Kansas, USA with five different serovars recovered. European sites were significantly more likely to yield Salmonella resistant to more than one antibiotic (OR 71.5, P < 0.001, CI = 3.77, 1358) than North American sites, where no resistance was found. Resistance to nalidixic acid, a quinolone, was recovered in nine isolates from four serovars in four different sites across Europe. Large corvids contribute to the transmission and dissemination of Salmonella and resistance genes between human and animal populations and across great distances. This information adds to the knowledge base of zoonotic pathogen prevalence and antibiotic resistance ecology in wild birds.     

Investigations of the Distribution and Persistence of Salmonella and Ciprofloxacin‐Resistant Escherichia coli in Turkey Hatcheries in the UK

This study aimed at gaining information on the presence of Salmonella in UK turkey hatcheries and possible epidemiological links between breeding farms, hatcheries and finishing farms. The presence of ciprofloxacin‐resistant E. coli in hatchery samples, as well as in faecal samples from farms, and trends in occurrence of resistance were also investigated. Over a 2 year‐period, four British turkey hatcheries were visited and intensively sampled for the presence of Salmonella and ciprofloxacin‐resistant E. coli. In two hatcheries, a link could be demonstrated between the presence of certain Salmonella serovars in the hatcheries and on breeding and finishing farms. Within the hatcheries, serovars linked to breeding farms were found more frequently in the poult processing and dispatch areas, whereas serovars identified as ‘resident hatchery contaminants’ were predominantly found inside the hatcher cabinets. Ciprofloxacin‐resistant isolates of S. Senftenberg were identified in one hatchery, which coincided with enrofloxacin treatment of some of the breeding flocks. Ciprofloxacin‐resistant E. coli was found in two hatcheries, and the majority of these isolates showed multidrug resistance.     

Characterization of Salmonella isolates from beef cattle, broiler chickens and human sources on Prince Edward Island

Non-typhoid Salmonella serovars remain a potential threat to human health, and beef cattle and broiler chickens are possible sources of these organisms on Prince Edward Island (PEI). In this study, the ceca of beef cattle belonging to fasted and non-fasted groups, and broiler chickens were examined for Salmonella at the time of slaughter. The characteristics of the isolates, including antimicrobial resistance patterns and virulence genes, were studied along with the isolates obtained from cases of human salmonellosis on PEI during the study period (1996–97). The prevalence of Salmonella in beef cattle was 4.6% (11/240). The rate was significantly higher in fasted cattle (7.46%), than in non-fasted cattle (0.94%). The prevalence rate in chickens was 32.5% (39/120). In beef cattle, Salmonella typhimurium phage type (PT) or definitive type (DT) 104 which was resistant to ampicillin, chloramphenicol, streptomycin, sulfisoxazole and tetracycline, was the most predominant type (64%). In chickens, S. heidelberg, with resistance to gentamicin, streptomycin and sulfisoxazole, predominated. Of 26 isolates from humans, the most common serovar was S. typhimurium, including a multidrug-resistant strain of DT104. Examination by PCR revealed presence of the virulence gene invA in all serovars, and the spvC gene in all S. typhimurium isolates, of both beef cattle and human origin. Among the other serovars the latter gene was found in 7 human isolates, but in none of the chicken or beef isolates. All but 3 of the spvC-positive isolates possessed a 90 kilobasepair (kbp) plasmid suggesting that the 3 isolates had the spvC gene on their chromosome. These findings were confirmed by plasmid DNA isolation using 3 different protocols and by sequence analysis of the spvC-PCR product.     

The occurrence of Salmonella,extended‐spectrum β‐lactamase producing Escherichia coli and carbapenem resistant non‐fermenting Gram‐negative bacteria in a backyard poultry flock environment

Increase in the number of small‐scale backyard poultry flocks in the USA has substantially increased human‐to‐live poultry contact, leading to increased public health risks of the transmission of multi‐drug resistant (MDR) zoonotic and food‐borne bacteria. The objective of this study was to detect the occurrence of Salmonella and MDR Gram‐negative bacteria (GNB) in the backyard poultry flock environment. A total of 34 backyard poultry flocks in Washington State (WA) were sampled. From each flock, one composite coop sample and three drag swabs from nest floor, waterer‐feeder, and a random site with visible faecal smearing, respectively, were collected. The samples were processed for isolation of Salmonella and other fermenting and non‐fermenting GNB under ceftiofur selection. Each isolate was identified to species level using MALDI‐TOFF and tested for resistance against 16 antibiotics belonging to eight antibiotic classes. Salmonella serovar 1,4,[5],12:i:‐ was isolated from one (3%) out of 34 flocks. Additionally, a total of 133 ceftiofur resistant (Cef^R) GNB including Escherichia coli (53), Acinetobacter spp. (45), Pseudomonas spp. (22), Achromobacter spp. (8), Bordetella trematum (1), Hafnia alvei (1), Ochrobactrum intermedium (1), Raoultella ornithinolytica (1), and Stenotrophomonas maltophilia (1) were isolated. Of these, 110 (82%) isolates displayed MDR. Each flock was found positive for the presence of one or more Cef^R GNB. Several MDR E. coli (n = 15) were identified as extended‐spectrum β‐lactamase (ESBL) positive. Carbapenem resistance was detected in non‐fermenting GNB including Acinetobacter spp. (n = 20), Pseudomonas spp. (n = 11) and Stenotrophomonas maltophila (n = 1). ESBL positive E. coli and carbapenem resistant non‐fermenting GNB are widespread in the backyard poultry flock environment in WA State. These GNB are known to cause opportunistic infections, especially in immunocompromised hosts. Better understanding of the ecology and epidemiology of these GNB in the backyard poultry flock settings is needed to identify potential risks of transmission to people in proximity.     

Epidemiological investigation of Leptospira spp. in a dairy farming enterprise after the occurrence of three human leptospirosis cases

An epidemiological investigation was conducted in an unvaccinated dairy farming enterprise in which three workers on one of the milking herds (Herd 1) were diagnosed with leptospirosis due to serovars Hardjo (H) (n = 2) and Pomona (P) (n = 1) between January and March 2015. Blood and urine samples were collected from milking cows in Herd 1 (N = 230) and Herd 2 (N = 400), rising one‐ (R1, N = 125) and rising two‐year‐old (R2, N = 130) replacement heifers, and four pigs associated with Herd 1, in March 2015. Sera were tested using the MAT for serovars H, P, Copenhageni (C), Ballum (B) and Tarassovi (T), and urine samples were tested by qPCR. Seventy‐five per cent of 109 cows in Herd 1 and 36% of 121 in Herd 2 were seropositive (≥48), predominantly to H and P, and 23% of 74 cows in Herd 1 and 1% of 90 cows in Herd 2 were qPCR positive. Fifty‐five per cent of 42 R2 heifers were seropositive to T. No R1 and 17% of 42 R2 heifers were qPCR positive. Subsequently, all cattle were vaccinated for H and P, and Herds 1 and 2 were given amoxicillin. After the booster vaccination, 7% of 91 in Herd 1, 2% of 82 in Herd 2 and 11% of 38 R1 heifers (sampled as R2) were PCR positive. After the amoxicillin treatment, no cows in Herd 1 and 5% of 62 cows in Herd 2 were urine PCR positive. Calves and pigs were seropositive to H, P, C and B. Vaccination and antibiotic treatment appeared effective in reducing the risk of exposure of workers to vaccine serovars. However, evidence of non‐vaccine serovars indicated that workers likely remain at risk of exposure to Leptospira.     

Multilocus Sequence Types of Environmental Campylobacter jejuni Isolates and their Similarities to those of Human,Poultry and Bovine C. jejuni Isolates

In this study, we investigated the multilocus sequence type (MLST) diversity and population genetics of Campylobacter jejuni isolates collected from the natural waters (n = 57), wild birds (n = 37) and zoo animals (n = 19) in southern Finland, the Helsinki area and the Helsinki Zoo, respectively. On average, we found C. jejuni in 20%, 10.4% or 11.5% of the samples collected from natural waters, wild birds and zoo animals, respectively. High ST diversity was detected in all three sources and 41.2% of the STs were novel, but the multi‐host adapted ST‐45 was the most common ST detected. The MLST data, supplemented with C. jejuni isolates from domestically acquired human infections (n = 454), poultry (n = 208) and bovines (n = 120), were utilized in a population structure study. The results indicate four groups of strains with varying ecological associations, demonstrating presence of genetically distinct lineages within each of the studied sources. We discovered that the greatest ST overlap occurs between human isolates and isolates from natural waters and poultry, which suggests that the latter two are the most important sources of C. jejuni among domestically acquired infections in Finland.     

Prevalence,antimicrobial resistance profile and comparison of selective plating media for the isolation of Salmonella in backyard chickens from Entre Rios,Argentina

This study was conducted to estimate the apparent prevalence of Salmonella spp. in birds kept under backyard system in Entre Ríos, Argentina, and determine the performance of two selective plating media used for Salmonella isolation, and the antimicrobial resistance of the isolated. Also, the association of farms characteristics with Salmonella presence was evaluated. A total of 657 backyard chickens and 15 gooses were sampled one time by cloacal swab, belonging to 51 and one family farms, respectively, and four counties in Entre Rios state from April 2014 to May 2015. Only four samples from backyard chickens belonged to three family farms from Uruguay County were positive to Salmonella spp., so the apparent prevalence was 0.6% for this kind of chicken. Four serovars were isolated (Salmonella ser. Lille, S. ser. Newport, S. ser. Enteritidis and S. ser. Rissen), which were susceptible to all antibiotics tested with the exception of erythromycin. For Hektoen enteric agar and brilliant green agar, relative specificity and positive predictive value were 1, and the relative sensitivity and negative predictive value did not show any difference between them. The agreement was very good between these two plating media. None of the variables studied could be selected to calculate the risk factors associated with Salmonella isolation because p > .15. Although the prevalence of Salmonella spp. is low in backyard birds in Entre Rios, the presence of S. ser. Enteritidis should not be discounted, because it is found in the county that concentrates a large population of intensive poultry production in the state.     

Prevalence and Antimicrobial Resistance Pattern of Salmonella Isolates from Apparently Healthy Slaughtered Cattle in Ethiopia

The prevalence and antimicrobial resistance pattern of Salmonella isolates was determined from apparently healthy slaughtered cattle at Debre Zeit (Ethiopia). A total of 323 cattle were examined for the presence of Salmonella in faeces, mesenteric lymph nodes, abdominal and diaphragmatic muscles. Salmonellae were cultured from 23 (7.1%) of the animals. Salmonellae were isolated from 2 (3.1%) and 3 (4.5%) of 65 pooled faecal and mesenteric lymph node samples, respectively. Nine (2.8%) abdominal muscle and 10 (3.1%) diaphragmatic muscle samples (n = 323 of each) were contaminated by Salmonella. About 60% of the serovars identified in the abdominal and diaphragmatic muscles were also detected from faeces and mesenteric lymph node samples. The five different serovars isolated were Salmonella mishmarhaemek (48%), S. typhimurium (20%), S. enteritidis (12%), S. guildford (12%) and S. dublin (48%). The antimicrobial resistance profiles of 25 of the Salmonella isolates with 17 antimicrobials showed that 52% (13/25) of them were resistant to three or more antimicrobials. Both strains of Salmonella (S. mishmarhaemek and S. typhimurium) showed multiple resistance to ampicillin, sulfamethoxazole and ticarcillin. This revised version was published online in July 2006 with corrections to the Cover Date.     

Prevalence of Multi-Drug-Resistant Salmonella in Equids Maintained by Low Income Individuals and on Designated Equine Farms in India

We examined 872 equids (445 maintained by low-income individuals and 427 maintained on nine designated equine farms) and, using previously described methods for bacteria, isolated Salmonella from fecal samples of 59 (6.77%) animals. Of the 646 horses, 183 donkeys, and 43 mules that had feces cultured for Salmonella, 42 (6.5%), 7 (3.8%), and 10 (23.3%), respectively, were excreting Salmonella strains in feces. Six horse mares were excreting Salmonella enterica of two different serovars simultaneously. A total of 65 Salmonella enterica isolates belonged to 13 serovars, namely S. paratyphi B var Java (14), S. I. 4, 5, 12, 27: r, i: 1, 5 (11), S. Drogana (8), S. Newport (7), S. Saintpaul (5), S. Lagos (4), S. Typhimurium (5), S. Kottbus (3), S. Bovismorbificans (3), S. Dumfries (2), S. Tshiongwe (1) S. Weltevreden (monophasic) (1), and S. enterica ssp salamae (1). With Salmonella-specific polymerase chain reaction (PCR) using hisJ gene primers, 107 (12.3) fecal samples yielded a specific amplicon of 496 bp. On using PCR, prevalence of Salmonella in donkeys, horses, and mules was 4.9%, 10.8%, and 65.1%, respectively. With both methods of Salmonella detection in feces, prevalence was significantly higher in female than in male donkeys and horses. Salmonella shedding in feces was significantly higher in equids maintained by low-income people than those at designated equine farms. Almost all Salmonella isolates (63 of 65) had multiple-drug-resistance (MDR, resistance to three or more drugs). Salmonella isolates were commonly resistant to sulfamethoxazole (90.8%), tetracycline (70.8%), doxycycline (67.7%), furazolidone (66.2%), and colistin (55.4%). A few isolates had resistance to trimethoprim (3.1%), ciprofloxacin (3.1%), ceftriaxone (3.1%), ceftazidime (3.1%), cefoperazone (3.1%), chloramphenicol (4.6%), cefotaxime (6.2%), gentamicin (9.2%), ampicillin + cloxacillin (9.2%), cotrimoxazole (13.8%), kanamycin (13.8%), amoxicillin + clavulanic acid (16.9%), imipenem (16.9%), ampicillin (18.5%), amikacin (23.1%), neomycin (27.7%), nalidixic acid (33.8%), and streptomycin (36.9%). With the exception of 13 Salmonella isolates of S. Drogana (4), S. Newport (4), S. I. 4, 5, 12, 27: r, i: 1, 5 (4) and S. Kottbus (1) serovars, all had one or more than one plasmid. Molecular weight of plasmids ranged between 3 kDa and >87 kDa. One heavy plasmid (≥87 kda) was present in all the 52 plasmid-positive strains. Presence of plasmid could not be correlated with MDR in Salmonella isolates from equids.     

Genome analysis of methicillin‐resistant Staphylococcus aureus isolated from pigs: Detection of the clonal lineage ST398 in Cameroon and South Africa

Food animals are considered reservoirs of methicillin‐resistant Staphylococcus aureus (MRSA) and are implicated in their zoonotic transmission in the farm‐to‐plate continuum. LA‐MRSA has been reported as a zoonotic agent that has the potential to spread to humans and may cause infections in at‐risk groups. In this study, whole genome sequencing was used to describe the genetic environment (resistance mechanisms, virulence factors and mobile genetic elements) and investigate the genetic lineages of MRSA isolates from pigs in Cameroonian and South African abattoirs. During March–October 2016, 288 nasal and rectal pooled samples from 432 pigs as well as nasal and hand swabs from 82 humans were collected. Genomic DNA was sequenced using an Illumina MiSeq platform. Generated reads were de novo‐assembled using the Qiagen CLC Genomics Workbench and SPAdes. The assembled contigs were annotated, and antibiotic resistance genes, virulence factors, plasmids, SCCmec and phage elements were identified with ResFinder, Virulence Finder, PlasmidFinder, SCCmec Finder and PHAST, respectively. Core genome single nucleotide analysis was undertaken to assess clonal relatedness among isolates. A lower MRSA prevalence was observed in pigs in Cameroon (n = 1/13; 0.07%) compared with South Africa (n = 4/22; 18.18%), and none of the workers were colonized by MRSA. Genome analysis identified various antibiotic resistance genes along with six virulence factors in all isolates. All MRSA isolates belonged to the clonal lineage ST398 (spa‐type t011) and harboured the type Vc SCCmec and several plasmids. Our study shows that the livestock‐associated MRSA clonal lineage ST398 is already present in both Cameroon and South Africa and is probably underestimated in the absence of molecular epidemiological studies. It reveals the serious food safety and public health threat associated with this animal strain and underscores the need for interventions to contain this resistant clone.     

A case–case study comparing the individual risk factors and symptomatology of Salmonella Heidelberg and Salmonella Typhimurium in Ontario in 2015, following implementation of the Ontario Investigation Tools

Salmonella Heidelberg and Salmonella Typhimurium are among the most common serotypes responsible for human salmonellosis in Ontario. Introduction of the Ontario Investigation Tools (OIT) in 2014 allowed for standardized case investigation and reporting. This study compared the risk factors and symptomatology for sporadic S. Heidelberg and S. Typhimurium cases reported in Ontario in 2015, following implementation of the OIT. Multilevel logistic regression models were applied to assess associations between serotype and individual‐level demographic characteristics, exposures and symptoms for sporadic confirmed cases of S. Heidelberg and S. Typhimurium in Ontario in 2015. There were 476 sporadic cases of S. Typhimurium (n = 278) and S. Heidelberg (n = 198) reported in Ontario in 2015. There were significant associations between the odds of the isolate from a case being one of these serotypes, and travel, consumption of sprouts (any type), contact with reptiles and development of malaise, fever or bloody diarrhoea. The S. Typhimurium and S. Heidelberg cases differed in both symptom presentation and risk factors for illness. Case–case comparisons of Salmonella serotypes have some advantages over case–control studies in that these are less susceptible to selection and recall bias while allowing for rapid comparison of cases to identify potential high‐risk exposures that are unique to one of the serotypes when compared to the other. Comparing cases of two different Salmonella serotypes can help to highlight risk factors that may be uniquely associated with one serotype, or more strongly associated with one serotype compared to another. This information may be useful for understanding relative source attribution between common serotypes of Salmonella.