Occurrence, Risk Factors, Serotypes, and Antimicrobial Resistance of Salmonella Strains Isolated from Imported Fertile Hatching Eggs, Hatcheries, and Broiler Farms in Trinidad and Tobago.

ABSTRACT: This cross-sectional study was conducted to determine the occurrence, risk factors, and characteristics of Salmonella isolates recovered from imported fertile broiler hatching eggs, hatcheries, and broiler farms in Trinidad and Tobago. Standard methods were used to isolate and characterize Salmonella isolates from two broiler hatcheries and 27 broiler farms in the country. The frequency of isolation of Salmonella was 0.0% for imported fertile hatching eggs (0 of 45 pools of 10 eggs each, i.e., 450 eggs), 7.6% for hatcheries (12 of 158 samples), and 2.8% for broiler farms (24 of 866 samples) (P = 0.006). Stillborn chicks at hatcheries had the highest prevalence of Salmonella (7 of 28 samples, 28.0%), whereas on broiler farms the cloacal swabs had the highest prevalence of Salmonella (15 of 675 samples, 2.2%). None of the 15 farm management and production practices investigated were significantly associated (P > 0.05) with the isolation of Salmonella. The predominant Salmonella serotypes were Kentucky (83.3%) and Infantis (62.5%) among hatchery and farm isolates, respectively. The disk diffusion method revealed frequencies of antimicrobial resistance (i.e., resistance to one or more agents) of 44.0% (11 of 25 isolates) and 87.5% (35 of 40 isolates) at hatcheries and broiler farms, respectively (P = 0.0002). Antimicrobial resistance among hatchery isolates was highest (28.0%) to doxycycline and kanamycin and was very high (>65%) among farm isolates to sulfamethoxazole-trimethoprim, gentamicin, ceftriaxone, kanamycin, and doxycycline. Multidrug resistance (MDR; i.e., resistance to antimicrobial agents from three or more classes) was exhibited by 4.0 and 85.7% of Salmonella isolates recovered from several environmental and animal sources at the hatcheries and farms, respectively (P < 0.0001). The high level of antimicrobial resistance and the presence of MDR among Salmonella isolates from broiler farms highlight the therapeutic implications and the potential for MDR strains to enter the food chain.

Characterization of Salmonella Isolates Recovered from Stages of the Processing Lines at Four Broiler Processing Plants in Trinidad and Tobago.

This cross-sectional study determined the prevalence, characteristics, and risk factors for contamination of chicken with Salmonella at four operating broiler processing plants in Trinidad. Standard methods were used to isolate and characterize the Salmonella isolates. The overall prevalence of Salmonella at the four processing plants was 27.0% (107/396). The whole carcass enrichment (WCE) method yielded a statistically significantly (p = 0.0014) higher frequency of isolation (53.9%; 97/180) than the whole carcass rinse (35.0%; 63/180) and neck skin methods (42.2%; 38/90). S. enterica serotypes Enteritidis, Javiana, and Infantis were the predominant serotypes isolated accounting for 20.8%, 16.7% and 12.5%, respectively, of the serotyped isolates. Risk factors included the use of over 100 contract farmers (OR 4.4), pre-chiller (OR 2.3), addition of chlorine to chiller (OR 3.2), slaughtering sick broilers (OR 4.4), and flocks with >50% mortality. Multi-drug resistance was detected in 12.3% (14/114) of the isolates of Salmonella. Resistance was high to kanamycin (85.7%) and doxycycline (74.6%) but low to amoxicillin-clavulanic acid (2.4%) and sulphamethoxazole-trimethoprim (0.8%). The occurrence of resistant Salmonella in chickens processed at commercial broiler processing plants has implications for salmonellosis and therapeutic failure in consumers of improperly cooked contaminated chickens from these plants in the country.

Prevalence of methicillin resistant Staphylococcus aureus (MRSA) in pigs and workers at abattoirs in Trinidad and Tobago.

INTRODUCTION: Methicillin resistant Staphylococcus aureus (MRSA), a major cause of zoonotic infections, has emerged globally in livestock, particularly pigs. People with occupational contact with food producing animals are at high risk of colonization. The aim of this study was to determine the prevalence of MRSA in pigs and abattoir workers throughout Trinidad and Tobago as well as their resistance to other antimicrobial agents. METHODOLOGY: Nasal and skin behind the ear swabs from pigs and nasal swabs from humans were enriched in Mueller Hinton broth with 6.5% sodium chloride, followed by phenol red mannitol broth with 75 mg/L aztreonam and 5 mg/L ceftizoxime. The enriched sample was then plated on both CHROMagar MRSA and Brilliance MRSA. All incubation was at 37ºC for approximately 24 h. Suspect MRSA isolates were confirmed as MRSA using the Penicillin-Binding Protein (PBP2a) test kit and polymerase chain reaction (PCR) to detect the mecA gene. Resistance of the S. aureus and MRSA isolates to 16 antimicrobial agents was determined using the disc diffusion method. RESULTS: Of the 929 pigs and 44 humans sampled, MRSA strains were isolated at a frequency of 0.9% (8/929) and 2.3% (1/44) respectively. All isolates exhibited resistance to one or more of the 16 antimicrobial agents. CONCLUSIONS: The study demonstrated that pigs and workers at slaughter houses in Trinidad and Tobago harbour multidrug resistance S. aureus and MRSA. This is of public health significance as occupational exposure of humans can lead to an increased risk of infection and therapeutic failure.

Prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in broilers and workers at 'pluck shops' in Trinidad.

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) is a cause of zoonotic infections in many countries. People with occupational contact with food animal production are at risk of colonization. The aim of this study was to determine the prevalence of MRSA and their frequency of resistance to other antimicrobial agents from broilers and workers at the 'pluck shops' in Trinidad. For isolation of MRSA, choanal, cloacal and pharyngeal swabs taken from broilers and nasal swabs from humans were enriched then plated on CHROMagar MRSA and Brilliance MRSA. MRSA was confirmed using the PBP2a test kit, resistance to oxacillin and cefoxitin and polymerase chain reaction (PCR) for the mecA gene. Antimicrobial resistance of the MRSA isolates to 16 antimicrobial agents was determined using the disc diffusion method. Of the 287 broilers and 47 humans sampled, MRSA was isolated at a frequency of 2 (0.7%) and 0 (0.0%) respectively. All the MRSA isolates exhibited resistance to one or more of the 16 antimicrobial agents. The study demonstrated that broilers at 'pluck shops' in Trinidad harbor MRSA. This is the first isolation of MRSA from poultry in Trinidad, West Indies, and this finding is of public health significance since occupational exposure of humans can lead to increased risk of acquiring MRSA infections.

Antimicrobial Resistance of Salmonella Isolates Recovered from Chickens Sold at Retail Outlets in Trinidad.

This study determined the frequency of resistance of 135 isolates of Salmonella, including 15 serotypes recovered from chickens purchased from retail outlets (cottage processors and supermarkets) across Trinidad. Resistance to 16 antimicrobial agents was determined by using the disk diffusion method. Resistance among the isolates was related to the type of retail outlet, location of outlets, type of sample, and isolate serotype. Overall, all isolates exhibited resistance to one or more of the 16 antimicrobial agents tested. All isolates were sensitive to cefoxitin and norfloxacin, with the overall frequency of resistance ranging from 1.1% (sulfamethoxazole-trimethoprim) to 100.0% (ceftiofur and doxycycline). The frequency of resistance to tetracycline, ampicillin, ceftriaxone, amoxycillin-clavulanic acid, and chloramphenicol was significantly ( P < 0.05) higher in isolates recovered from cottage processor outlets compared with those from supermarkets. The frequency of resistance to antimicrobial agents was significantly different only to kanamycin ( P = 0.046) and enrofloxacin ( P = 0.000) across seven counties in Trinidad). Regarding sample presentation (whole versus parts), the frequency of resistance was only significantly higher to gentamicin ( P = 0.039) for chicken part isolates from cottage processor and to only tetracycline ( P = 0.034) for isolates from whole carcasses from supermarkets. All the 135 Salmonella isolates exhibited multidrug resistance patterns. The high frequency of resistance to seven antimicrobial agents (erythromycin, streptomycin, ceftiofur, doxycycline, kanamycin, tetracycline, and ampicillin), some used in the poultry industry, coupled with the occurrence of multidrug resistance, may have potential therapeutic implications for broiler farmers in Trinidad.

Prevalence and serotypes of Salmonella spp. on chickens sold at retail outlets in Trinidad.

BACKGROUND: This cross-sectional study determined the prevalence of Salmonella spp. and their serotypes on dressed chicken sold at retail outlets in Trinidad. The study also investigated the risk factors for contamination of dressed carcasses by Salmonella spp. at cottage poultry processor outlets where chickens are slaughtered and processed for sale. METHODS: A total of 133 dressed, whole chickens and 87 chicken parts from 44 cottage poultry processors and 36 dressed, whole chickens and 194 chicken parts from 46 supermarket outlets were randomly collected throughout the country. Isolation and identification of Salmonella spp. were performed using standard bacteriological techniques. Serotyping was performed by a regional reference laboratory. RESULTS: The prevalence of Salmonella spp. in chicken carcasses sampled from cottage poultry processors and supermarkets was 20.5% and 8.3% respectively (p <0.001). The frequency of isolation of Salmonella spp. at cottage poultry processors was 22.4%, 23.0%, 7.1%, and 10.0% for non-chilled whole chicken, non-chilled chicken parts, chilled whole chicken and chilled chicken parts respectively. Fresh, non-chilled chicken (22.6%) yielded a higher frequency of isolation of Salmonella spp. than chilled chickens (8.3%). For supermarket samples, the frequency of isolation of Salmonella spp. was 19.0%, 8.1%, 0.0% and 7.6% for chilled whole chickens, chill chicken parts, frozen whole chicken and frozen chicken parts respectively. The swab method of sampling yielded a statistically significantly (p = 0.029) higher frequency (3.2%) of Salmonella spp. than the rinse method (1.6%). The predominant serotypes isolated were Kentucky (30.9%) and Javiana (22.7%). Use of chilled water-bath to cool carcasses was the only risk factor significantly (p = 0.044) associated with isolation of Salmonella spp. CONCLUSION: Raw chicken carcasses purchased from cottage poultry processors pose a significantly higher risk of contamination with Salmonella spp. than those sold at supermarkets.

Serological evidence of hantavirus infection in farm and abattoir workers in Trinidad--a preliminary study.

Hantaviruses are established causative agents of hemorrhagic fevers and renal diseases amongst other clinical manifestations in humans, with most diagnosis based on serological assays. The disease, which is rodent-borne, has been reported in numerous countries worldwide but information about the disease is scanty in the Caribbean. The objective of this investigation is to determine the frequency of exposure to hantaviruses in a selected apparently healthy human population associated with abattoirs and livestock farms in Trinidad using a hantavirus immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA). Of a total of 236 individuals tested, 27 (11.4%) were seropositive for hantavirus infection. Amongst abattoir workers the frequency of infection was 9.4% (6 of 64) compared with seropositivity rate of 12.4% (18 of 145) and 11.1% (3 of 27) amongst livestock farm workers and office workers and other individuals with minimal animal contact respectively. The differences were, however, not statistically significant (p > .05; χ(2) test). Age, gender, and race did not significantly affect the infection rate by hantavirus in the workers studied. This is considered the first documented evidence of hantavirus infection in Trinidad and Tobago. It is imperative for local physicians to consider hantavirus as a differential diagnosis in patients with hemorrhagic fever and renal diseases, since there may be a number of undiagnosed cases of hantavirus disease in the human population in the country.

Frequency of detection of immunoglobulins of Toxoplasma gondii, Leptospira spp., and Brucella abortus in livestock/farm and abattoir workers in Trinidad.

Toxoplasma gondi, Leptospira spp., and Brucella abortus are all established parasitic and bacterial zoonoses that manifest themselves in several forms of human diseases. They have been associated with occupational exposures, particularly amongst workers associated with livestock farms. The primary objectives of this study were to determine the seroprevalence of T. gondii immunoglobulin M (IgM) immunoglobulins (serum antibodies), Leptospira IgM immunoglobulins, and B. abortus IgG immunoglobulins, suggestive of acute or chronic infections, in livestock/farm and abattoir workers in Trinidad, and to relate to risk factors. Blood samples were collected from 394 consenting livestock/farm workers and 99 abattoir workers across the island of Trinidad. Serological status was determined by enzyme immunoassay (EIA) for T. gondii IgM, enzyme-linked immunosorbent assay (ELISA) for Leptospira IgM immunoglobulins, and buffered plate agglutination test (BPAT) and competitive ELISA for B. abortus IgG immunoglobulins. Of 394 apparently healthy livestock/farm workers sampled, 150 (38.1%) were seropositive for T. gondii IgM immunoglobulins, compared with 44 (44.4%) of 99 abattoir workers (p > .05; χ(2) test). Five (1.3%) of 371 and 1 (1.0%) of 99 livestock/farm and abattoir workers respectively were positive for Leptospira IgM immunoglobulins. All samples from livestock/farm workers and abattoir workers were negative for B. abortus immunoglobulins. None of the risk factors investigated was statistically significantly (p > .05; χ(2) test) associated with T. gondii and Leptospira spp. infections.

Frequency of seropositivity for Coxiella burnetii immunoglobulins in livestock and abattoir workers in Trinidad.

Coxiella burnetii is a zoonotic, rickettsial pathogen which causes mild and severe diseases often referred to as Q-fever in humans, particularly those occupationally exposed. This study determined the seropositivity for Coxiella burnetii IgM immunoglobulins using the enzyme immunoassay (EIA) in livestock and abattoir workers in Trinidad and related to selected personal characteristics to seroprevalence. Overall, of the 455 humans whose serum samples were tested, 20 (4.4%) were seropositive for C. burnetii IgM immunoglobulin, comprising 13 (4.6%) out of 283 livestock workers, 4 (4.7%) out of 85 abattoir workers and 3 (3.4%) out of 87 office workers (P>0.05; Chi 2). The age, sex and race of workers were not significantly associated with the occurrence of acute Q-fever (P>0.05; Chi 2). This is considered the first documentation of Q-fever in the human population in Trinidad. It is difficult to assess the impact of the disease in the country since the disease is not routinely tested for in the local hospitals or diagnostic laboratories.

Frequency of seropositivity for Coxiella burnetii immunoglobulins in livestock and abattoir workers in Trinidad

Coxiella burnetii is a zoonotic, rickettsial pathogen which causes mild and severe diseases often referred to as Q-fever in humans, particularly those occupationally exposed. This study determined the seropositivity for Coxiella burnetii IgM immunoglobulins using the enzyme immunoassay (EIA) in livestock and abattoir workers in Trinidad and related to selected personal characteristics to seroprevalence. Overall, of the 455 humans whose serum samples were tested, 20 (4.4%) were seropositive for C. burnetii IgM immunoglobulin, comprising 13 (4.6%) out of 283 livestock workers, 4 (4.7%) out of 85 abattoir workers and 3 (3.4%) out of 87 office workers (P>0.05; ö2).The age, sex and race of workers were not significantly associated with the occurrence of acute Q-fever (P>0.05; ö2). This is considered the first documentation of Q-fever in the human population in Trinidad. It is difficult to assess the impact of the disease in the country since the disease is not routinely tested for in the local hospitals or diagnostic laboratories.

Investigation for possible source(s) of contamination of ready-to-eat meat products with Listeria spp. and other pathogens in a meat processing plant in Trinidad.

In 2003, there was a recall of three processed (chicken franks, spice ham and turkey ham ready-to-eat (RTE) meat products by a large processing plant in Trinidad as a result of contamination by Listeria monocytogenes. The study was conducted to investigate the possible source(s) of Listeria contamination of recalled RTE meat products and to determine the prevalence of Listeria spp., Salmonella spp., Escherichia coli and Campylobacter spp. in the products and air within the plant. Raw and processed meat products, as well as food contact surfaces were also tested for Salmonella spp., Listeria spp. and Campylobacter spp. initially after thorough clean-up and close-down of the plant. Faecal and effluent samples from the piggery, in close proximity to the plant, were tested for the presence of Salmonella spp., Listeria spp. and Campylobacter spp. Air samples and food contact surfaces were negative for the tested organisms. Ten (58.8%) of the 17 effluent samples and 4 (11.8%) of the 34 faecal samples were positive for Campylobacter coli. Of the 11 raw meat products tested, 10 (90.9%) were positive for E. coli and Listeria spp. either singly or in combination. Of the 32 processed RTE products tested, 11 (34.4%) were positive for E. coli, Salmonella spp., Listeria spp. and Campylobacter spp. in combination or singly. Eleven (61.1%) of 18 processed products contained unacceptable levels of aerobic bacteria using international standards. Four months later, following the implementation of recommended cleaning, sanitizing and hygienic practices at the plant, pre- and post-processed products were sampled and Listeria spp. were identified in 4 (80.0%) of the 5 raw products and in 1 of the 5 (20.0%) finished products. Two (40.0%) of the finished products contained unacceptable microbial levels. It was concluded that the close proximity of the piggery to the processing plant was not the probable source of Listeria contamination of the recalled meat products. The data suggested that improved sanitary practices on food contact surfaces and during handling of products, reduced the risk of Listeria spp. and other pathogens studied. The problem at the plant can therefore, be inferred to be due to lapses in good sanitary practices, inadequate heat treatments or the presence of pathogens particularly Listeria in biofilms on different surfaces continuously or occasionally contaminating finished products.

Investigation for possible source(s) of contamination of ready-to-eat meat products with Listeria spp. and other pathogens in a meat processing plant in Trinidad

In 2003, there was a recall of three processed (chicken franks, spice ham and turkey ham ready-to-eat (RTE) meat products by a large processing plant in Trinidad as a result of contamination by Listeria monocytogenes. The study was conducted to investigate the possible source(s) of Listeria contamination of recalled RTE meat products and to determine the prevalence of Listeria spp., Salmonella spp., Escherichia coli and Campylobacter spp. in the products and air within the plant. Raw and processed meat products, as well as food contact surfaces were also tested for Salmonella spp., Listeria spp. and Campylobacter spp. initially after thorough clean-up and close-down of the plant. Faecal and effluent samples from the piggery, in close proximity to the plant, were tested for the presence of Salmonella spp., Listeria spp. and Campylobacter spp. Air samples and food contact surfaces were negative for the tested organisms. Ten (58.8 per cent) of the 17 effluent samples and 4 (11.8 per cent) of the 34 faecal samples were positive for Campylobacter coli. Of the 11 raw meat products tested, 10 (90.9 per cent) were positive for E. coli and Listeria spp. either singly or in combination. Of the 32 processed RTE products tested, 11 (34.4 per cent) were positive for E. coli, Salmonella spp., Listeria spp. and Campylobacter spp. in combination or singly. Eleven (61.1 per cent) of 18 processed products contained unacceptable levels of aerobic bacteria using international standards. Four months later, following the implementation of recommended cleaning, sanitizing and hygienic practices at the plant, pre- and post-processed products were sampled and Listeria spp. were identified in 4 (80.0 per cent) of the 5 raw products and in 1 of the 5 (20.0 per cent) finished products. It was concluded that the close proximity of the piggery to the processing plant was not the probable source of Listeria contamination of the recalled meat products.