Akram R. Alaboudi

Occurrence and Antimicrobial Susceptibility of Listeria monocytogenes Isolated from Brined White Cheese in Jordan

Listeria monocytogenes is a serious foodborne pathogen that has been isolated from different dairy food products. Several foodborne outbreaks of listeriosis have been associated with consumption of cheese. The aims of this study were to determine the occurrence of L. monocytogenes and Listeria spp. in brined white cheese (BWC) sold in Jordan, and to determine the susceptibility of isolated L. monocytogenes to antimicrobials. Three hundred and fifty samples of 5 different types of BWC (akkawi, boiled, halloumi, pasteurized, and shellal) were collected from a local market in Jordan. The ISO (11290-1) procedure was followed for isolation and identification of Listeria spp. from cheese samples and a polymerase chain reaction (PCR) technique was used for confirmation of L. monocytogenes isolates. The VITEK2 automated system was used for testing antimicrobial susceptibility of L. monocytogenes isolates. The overall prevalence of Listeria spp. in cheese sample was 27.1%. L. monocytogenes was isolated from 39 (11.1%) samples. Other isolated species were L. grayi (6.9%), L. innocua (2%), L. ivanovii (4%), L. seeligeri (2%), and L. welshimeri (0.3%). The pH values and salt concentrations of L. monocytogenes positive cheese samples ranged from 5.10 to 6.32 and 5.64 to 13.16, respectively. L. monocytogenes isolates were sensitive or intermediate susceptible to imipenem, gentamicin, linezolid, teicoplanin, vancomycin, fusidic acid, trimethoprim/sulfamethoxazole, benzylpenicillin, ciprofloxacin, erythromycin, tetracycline, and rifampicin, but resistant to fosfomycin, oxacillin, and clindamycin.

Prevalence and antimicrobial susceptibility of Escherichia coli O157:H7 on beef cattle slaughtered in Amman abattoir

Cattle are the main asymptomatic reservoir of Escherichia coli O157:H7 which can cause illness to human. The objectives of the study were to measure the prevalence of E. coli O157:H7 on cattle slaughtered in Amman abattoir, detect virulence factors in the isolates, determine antibacterial resistance of the isolates, and know how the isolates are different or similar when compared to characterized isolates from developed countries. A total of 540 samples (feces, hide, and carcass) were tested for E. coli O157:H7 using the method of ISO 16654:(E). Conventional and multiplex PCR assays were used for serotype confirmation and virulence factor detection, respectively. Fifty E. coli O157:H7 isolates were identified and virulence factors eaeA and hlyA were present in all of the isolates. 60%, 12%, and 22% of the isolates harbored stx(1), stx(2), and stx(1) and stx(2), respectively. The prevalence rates of enterotoxigenic E. coli O157:H7 (n=47) were 8.3%, 10%, and 7.8% in feces, hides and carcasses, respectively. The antimicrobial profiles of the isolates showed an extensive resistance to erythromycin, neomycin and vancomycin and high sensitivity to ampicillin, ciprofloxacin, gentamicin, kanamycin and tetracycline.

Prevalence and antimicrobial susceptibility of Campylobacter spp. in live and dressed chicken in Jordan.

A total of 140 broiler flocks presented for slaughtering at Amman slaughterhouse were tested for Campylobacter spp. via collection of cloacal swabs from live birds, feathered skin samples at prescalding, and skin samples at postscalding (62°C or 57°C scalding temperature), postevisceration, and postchilling. The results indicated that 40% of the flocks tested by cloacal swabs, 34% at prescalding, 32% at post 57°C scalding, and 32% postevisceration were harboring Campylobacter jejuni. None of the skin samples collected from dressed birds at postscalding (62°C) or postwashing-chilling steps (regardless of scalding temperature) revealed the presence of C. jejuni. Thirty eight isolates were tested for susceptibility to ten antimicrobials by using the microbroth dilution method. Almost 50% of the isolates were multidrug resistant to 9 or 10 out of the ten tested antimicrobials. The other half of tested isolates were sensitive to erythromycin, tetracycline, doxycyclin, chlortetracycline, ciprofloxacin, enorfloxacin, gentamycin, tilmicosin, amoxicillin, and trimethoprim.

Detection, Identification, and Prevalence of Pathogenic Vibrio parahaemolyticus in Fish and Coastal Environment in Jordan.

Vibrio parahaemolyticus is widely distributed in the marine environments and considered the leading cause of human gastroenteritis in Asian countries. A total of 150 marketed fish and 50 water and sediment samples from the Gulf of Aqaba were examined for the prevalence of pathogenic strains of V. parahaemolyticus. A total of 132 typical isolates obtained from the primary selective medium (thiosulfate-citrate bile salt sucrose agar) and showed positive biochemical properties were subjected to confirmation by polymerase chain reaction targeting the gyrB and toxR genes. These genes were confirmed at rates of 82% (108 isolates) and 72% (95 isolates), respectively. The toxR positive isolates were tested for the presence of thermolabile hemolysin (tlh), thermostable direct hemolysin (tdh), and tdh-related hemolysin (trh) virulence genes. Accordingly, the prevalence rates of pathogenic V. parahaemolyticus were 4%, 8%, and 12% in sediment, water, and fish samples, respectively. The 16S rRNA amplification and sequences were conducted for confirmation of the isolates and showing the relatedness among these isolates. The results showed that both 16S rRNA and toxR assays had same sensitivity and tested isolates had high nucleotide similarity irrespective of their sources.

Feasibility of Using Gamma Irradiation for Inactivation of Starvation-, Heat-, and Cold-Stressed Salmonella in Tahini

Salmonella continues to be the leading cause of acute gastroenteritis and recently has been involved in infections related to edible seeds and their products, including tahini. This study investigated the (i) effectiveness of using gamma irradiation to inactivate starvation- and heat- or cold-stressed Salmonella in tahini, (ii) effect of storage on the sensitivity of stressed Salmonella to irradiation, and (iii) effect of irradiation on the chemical and physical characteristics of tahini. Tahini samples were inoculated with a cocktail of unstressed or stressed (starvation and heat or cold stress) Salmonella isolates and then exposed after storage at 21°C for 0, 7, and 30 days to gamma irradiation for up to 2.0 kGy. Additionally, the effect of irradiation on the color, peroxide, p-anisidine, and acid values of tahini were assessed. The initial level of unstressed and starvation- and heat-stressed Salmonella in tahini decreased by ca. 4.6 log CFU/g after exposure to 2.0 kGy, while cold-stressed cultures decreased by 4.5 log after exposure to 0.6 kGy. Irradiation doses of 1.0 kGy after 7 days of storage or 0.75 kGy after 30 days of storage decreased the populations of the unstressed and starvation- and heatstressed Salmonella by ca. 3.4 or 2.6 log, respectively. The D10-value of the unstressed Salmonella was 0.43 kGy. Starvation and heat stresses showed no significant effect (P > 0.05) on the calculated D10-value, whereas cold stress significantly (P < 0.05) decreased the D10-value to 0.14 kGy. Preirradiation storage for 7 and 30 days significantly decreased the D10-value to 0.31 and 0.28 kGy, respectively. An irradiation dose of 2.0 kGy did not significantly affect the color, peroxide, p-anisidine, and acid values of tahini when compared with nonirradiated samples. Therefore, this study lays the foundation for using irradiation as an effective means for minimizing the risk of Salmonella in tahini without compromising its quality.

Survival and growth of Salmonella Typhimurium, Escherichia coli O157:H7 and Staphylococcus aureus in eggplant dip during storage

Eggplant dip is an internationally popular appetizer, prepared in some instances under uncertain hygienic conditions with inconsistent refrigeration. This study examined the effects of citric acid on the survival of pathogenic microorganisms (Salmonella Typhimurium, Escherichia coli O157:H7 and Staphylococcus aureus) and naturally present organisms (lactic acid bacteria, LAB, aerobic bacteria, APC and yeast and mold, YM) in eggplant dip during storage. Eggplant dip with 0, 0.2, 0.4, 0.6 or 0.8% citric acid was inoculated with S. Typhimurium, E. coli O157:H7 or S. aureus and stored at 4, 10 and 21 °C for ≤15 d. Throughout the study, the survival of the inoculated microorganisms was monitored, and LAB, APC, YM numbers and pH were determined. There was no significant (p>0.05) effect of citric acid on inoculated S. Typhimurium and E. coli O157:H7. Salmonella and E. coli O157:H7 survived >7d with little reduction in viability. Reduction of S. aureus viability increased with citric acid concentration and reached>3.0 log10 CFU/g by 15 d at 4 °C. Citric acid had no effect (p>0.05) on the background YM during storage at 4, 10 and 21 °C or LAB stored at 4 and 10 °C, while at 21 °C, 0.6 and 0.8% citric acid significantly reduced LAB. Citric acid had no effect (p>0.05) on the APC in samples stored at 4 °C but it had significant effects on samples stored at 10 and 21 °C. Work reported showed that the use of citric acid at 0.4-0.8% can inhibit the growth of S. aureus in eggplant dip, but adequate refrigeration is essential to minimize risk from this and other pathogens in this product.

Escherichia coli O157:H7 Facilitates the Penetration of Staphylococcus aureus into Table Eggs

Fresh eggshells collected from a local farm were subjected to different levels of surface contamination with feces containing different levels (3 to 5 log₁₀) of Escherichia coli O157:H7 or Staphylococcus aureus and incubated at 3 different temperatures (10, 25, and 32 °C). The penetration rates of contaminating bacteria were followed throughout the incubation period by tracing bacterial presence in shell, shell membranes, albumen, and yolk. The study revealed the ability of both E. coli O157:H7 and enterotoxigenic S. aureus to grow on shell in feces, penetrate the shell, and move and multiply within egg contents at different rates and periods depending on bacterial type and incubation conditions. High temperatures (25 and 32 °C) increased penetration rate, whereas storage at 10 °C decreased significantly the rate of penetration. High levels of contamination with E. coli O157:H7 also shortened the time needed for the penetration process. Results showed that when eggshells were contaminated with both organisms simultaneously, the penetration of E. coli O157:H7 preceded that of S. aureus and facilitated the invasion of the latter bacteria.

Biotypes and enterotoxigenicity of staphylococci isolated from camel's meat in Jordan.

A total of 264 camel’s meat and nasal swab samples were collected for isolation and typing of Staphylococci from Irbid Governorate in northern Jordan. About 97 % and 85% of meat and nasal swabs samples showed typical colonies of Staphylococcus aureus on BairdParker agar respectively. Out of 243 presumptively identified isolates, only 74 and 64 were confirmed as S. aureus by Microbact system and PCR technique respectively. About 67% of the isolates were typable by Devriese’s scheme. Fifteen of those isolates (23%) were specifically allocated to human, bovine, ovine or abattoir where, 14% of these host specific isolates belonged to human biovar. The other 44% belonged to non-host specific biovars with majority of them were allocated to NHS1 biovar. When tested for the presence of toxin genes, 71.9% of S. aureus isolates had SE(s) genes with SEA being the most prominent at 91.3%. The study also showed that not only coagulase positive isolates contain toxin genes, coagulase negative isolates also possess toxin genes and thus are considered potential hazards in camel’s meat.

Decontamination and survival of Enterobacteriaceae on shredded iceberg lettuce during storage

Enterobacteriaceae family can contaminate fresh produce at any stage of production either at pre-harvest or post-harvest stages. The objectives of the current study were to i) identify Enterobacteriaceae species on iceberg lettuce, ii) compare the decontamination efficiency of water, sodium hypochlorite (free chlorine 200 ppm), peroxyacetic acid (PA 80 ppm; Kenocid 2100®) or their combinations and ionizing radiation against Enterobacteriaceae on shredded iceberg lettuce and iii) determine the survival of Enterobacteriaceae post-treatment storage of shredded iceberg lettuce at 4, 10 and 25 °C, for up to 7 days. Klebsiella pneumonia spp. pneumonia, Enterobacter cloacae, Klebsiella oxytoca, Pantoea spp., Leclercia adecarboxylata and Kluyvera ascorbate were identified on iceberg lettuce. No significant difference (P≥ 0.05) among Enterobacteriaceae survival after washing with water or sanitizing with sodium hypochlorite or Kenocid 2100® (reduction ≤ 0.6 log CFU/g) were found. Combined sanitizer treatments were more effective against Enterobacteriaceae than single washing/sanitizing treatments. Sanitization of iceberg lettuce with combined washing/sanitizing treatments reduced Enterobacteriaceae by 0.85-2.24 CFU/g. Post-treatment growth of Enterobacteriaceae during storage on samples sanitized with sodium hypochlorite and Kenocid 2100® was more than on samples washed with water. The D10-value of Enterobacteriaceae on shredded iceberg lettuce was 0.21 KGy. The reduction of Enterobacteriaceae populations on iceberg after gamma radiation (0.6 KGy) was 3 log CFU/g, however, Enterobacteriaceae counts increased post-irradiation storage by 4-5 log CFU/g. Therefore, washing shredded iceberg lettuce with combined sanitizing treatment (sodium hypochlorite/sodium hypochlorite, sodium hypochlorite/Kenocid 2100®, or Kenocid 2100®/Kenocid 2100®) for total time of 6 min or exposing it to gamma irradiation (0.6 KGy) can decrease the risk of Enterobacteriaceae (reduction ≥ 2 log). Post-washing storage of sliced iceberg lettuce (4, 10, 25 °C) could increase the risk of Enterobacteriaceae as their counts increased during storage even at low temperatures.

Antimicrobial Residues in Table Eggs

Veterinary drugs are used in laying hens for therapeutic reasons. Antimicrobial residues can accumulate in egg components when administered to laying hens or when laying hens are mistakenly given medicated feed, or when the diet of egg-laying strains of chickens is accidently contaminated at the feed mill due to prior use of antimicrobials with other feeds. Misuse of antimicrobials through improper licensing and unwatched withdrawal periods are commonly observed in developing countries. When these chemicals reach the hen’s blood, these antimicrobials are deposited into egg yolk and albumen depending on their water or lipid solubility and protein binding ability. Egg processing through heating or storage reduces the levels of these antimicrobials but not to levels considered safe for human consumption. Antimicrobials fed to poultry are an important public health issue because of possible pathogenic microbial resistance. Routine surveillance for antimicrobials in eggs should be implemented.