Occurrence, virulence, and resistance genes in Salmonella enterica isolated from an integrated poultry company in Jordan.

Salmonella is considered one of the most common foodborne pathogens worldwide. The annual number of hospitalizations and deaths related to zoonotic salmonellosis, which is transmitted from animals to humans and infects poultry and meat, is expected to be significant. Hence, the primary aims of this research were to isolate and characterize Salmonella species obtained from an integrated poultry company and identify some virulence, and antimicrobial resistance, with a specific concern about colistin resistance genes. A total of 635 samples collected from various sources in an integrated company in Jordan were screened for Salmonella species accompanying their virulence and antimicrobial resistance genes. Samples were collected from parent stock house drag swabs, broiler farms, premix, cecum at the slaughterhouse level, prechilling and postchilling stages, and the final product. Salmonella species were detected in 3% (6/200) of investigated parent stock house drag swabs, 13.8% (11/80) from cloacal swabs from broiler farms, 16.9% (11/65) from boiler farms premix, 24.4% (11/45) from the cecum at slaughterhouse level, 16.4% (9/55) from the prechilling stage, 37.8% (17/45) from the postchilling stage and 53.3% (24/45) from the final product stage. No isolates were detected in feed mills (0/20), parents' premix (0/40), or hatcheries (0/40). Salmonella isolates were resistant to ciprofloxacin (91.0%), nalidixic acid (86.5%), doxycycline (83.1%), tetracycline (83.1%), sulphamethoxazole-trimethoprim (79.8%) and ampicillin (76.4%). Serotyping shows that S. Infantis was the predominant serovar, with 56.2%. Based on the minimum inhibitory concentration (MIC) test, 39.3% (35/89) of the isolates were resistant to colistin; however, no mcr genes were detected. Among antimicrobial-resistant genes, blaTEM was the most prevalent (88.8%). Furthermore, the spvC, ompA, and ompF virulence genes showed the highest percentages (97.8%, 97.8%, and 96.6%, respectively). In conclusion, Salmonella isolates were found at various stages in the integrated company. S. Infantis was the most prevalent serotype. No mcr genes were detected. Cross-contamination between poultry production stages highlights the importance of good hygiene practices. Furthermore, the presence of virulence genes and the patterns of antimicrobial resistance present significant challenges for public health.

Risk factors associated with mcr-1 colistin-resistance gene in Escherichia coli broiler samples in northern Jordan.

OBJECTIVES: The purpose of this study was to determine the prevalence of colistin-resistant Escherichia coli carrying mcr-1, and to identify risk factors associated with mcr gene-mediated resistance. METHODS: In total, 385 cloacal samples were collected from 125 broiler farms and a questionnaire containing information about each farm was designed and filled. RESULTS: Most of the antibiotics used in the disk diffusion method were highly resistant in all samples, with tetracycline and penicillin showing 100% and 99.7% resistance, respectively. Additionally, avian pathogenic E. coli (APEC) virulence genes frequency and percentage of APEC were identified, including sitA,iucC, and astA at 77%, 70.5%, and 62% respectively. In total, 214 of 360 isolates were positive for APEC (59.4%). Based on the minimum inhibitory (MIC) test, 58% of the isolates (n = 209 of 360) were resistant to colistin, with 39.7% displaying the mcr-1 gene. The statistical analysis of risk factors that influence colistin resistance prevalence revealed several significant factors, including commercial feed, farm management, sanitization, and antibiotic use. Irregular health checks for workers, non-dipping of feet before entering poultry houses, and the use of commercial poultry feeds all contributed to higher levels of colistin resistance as measured by MIC. On the other hand, doxycycline and commercial feed was 4 and 3.2 times more likely to occur based on the final logistic model of the mcr-1 gene, respectively. CONCLUSION: Our results suggest that better biosecurity protocols should be implemented in poultry farms to reduce antibiotic-resistant bacteria. Additionally, antibiotics should be carefully monitored and used only when necessary.

Multi-drug Resistant Acinetobacter baumannii: Phenotypic and Genotypic Resistance Profiles and the Associated Risk Factors in Teaching Hospital in Jordan.

BACKGROUND: This study aimed to determine the prevalence of Antimicrobial Resistance Genes (ARGs), with a focus on colistin resistance in clinical A. baumannii, as well as the risk factors associated with A. baumannii infection in Jordanian patients. METHODS: In total, 150 A. baumannii isolates were obtained from patients at a teaching hospital. The isolates were tested for antimicrobial susceptibility using disc diffusion and microdilution methods. PCR amplification was used to detect ARGs, and statistical analysis was conducted to evaluate the influence of identified risk factors on the ARGs acquisition. RESULTS: More than 90% of A. baumannii isolates were resistant to monobactam, carbapenem, cephalosporins, Fluoroquinolones, penicillin, and ß-lactam agents. Moreover, 20.6% of the isolates (n = 31) were colistin-resistant. Several ARGs were also detected in A. baumannii isolates. Univariate analysis indicated that risk factors and the carriage of ARGs were significantly associated P ≤ (0.05) with gender, invasive devices, immunodeficiency, systemic diseases, tumors, and covid-19. Logistic regression analysis indicated seven risk factors, and three protective factors were associated with the ARGs (armA, strA, and strB) P ≤ (0.05). In contrast, tetB and TEM were associated with 2 risk factors each P ≤ (0.05). CONCLUSION: Our study indicates a high prevalence of MDR A. baumannii infections in ICU patients, as well as describing the case of colistin-resistant A. baumannii for the first time in Jordan. Additionally, the risk factors associated with ARGs-producing A. baumannii infections among ICU patients suggest a rapid emergence and spread of MDR A. baumannii without adequate surveillance and control measures.

Emergence of mcr-1 gene and carbapenemase-encoding genes among colistin-resistant Klebsiella pneumoniae clinical isolates in Jordan.

BACKGROUND: Antimicrobial resistance (AMR) is a global threat that requires serious attention, particularly when it is developed against colistin, which is considered one of the 'last-resort' antibiotics for curing an infection. This study aimed to investigate the AMR profile of the Klebsiella (K.) pneumoniae clinical isolates and to obtain the comprehensive characteristics of the carbapenemases among the carbapenem-resistant K. pneumoniae (CR-KP) when isolated. In addition, to detect the colistin resistance and investigate the MCR genes in the clinical K. pneumoniae isolates for the first time in Jordan. METHODS: A total of 179 K. pneumoniae isolates were cultured and they were confirmed using the VITEK 2 system and PCR. The antibiotic susceptibilities, extended-spectrum beta-lactamase (ESßL), multidrug-resistant (MDR), and CR-KP were determined by using the VITEK 2 system, disc diffusion, and the minimum inhibitory concentration (MIC) test. PCR was performed to detect the MCR and carbapenemase genes. RESULTS: The rates of ESßL, MDR, and CR-KP were 48 %, 62 %, and 12.8 %, respectively. High colistin resistance of 49.7 % (89/179) was found. Only one MCR-1 (1.1 %) out of the 89 colistin resistance isolates was detected. Many of the isolates harbored the ESßL genes. In particular, the carbapenem genes were detected in 26 isolates, with 46 % KPC enzyme genes (12/26), 23 % IMP genes (6/26), 19 % OXA-48 genes (5/26), 11.5 % NDM-1 genes (3/26) but no VIM gene was found. The statistical analyses revealed a significant association between colistin resistance and MDR (P ≤ 0.05, Chi-square test). An association between colistin resistance and the Piperacillin, Ceftazidime, Cefpodoxime, Imipenem, Aztreonam, and Tobramycin resistance was noted. CONCLUSION: The study's findings demonstrated the presence of the MCR-1 gene in the K. pneumoniae clinical isolates for the first time in Jordan and indicated that the KPC and IMP encoded carbapenemases were the most prevalent K. pneumoniae carbapenemases in Jordan patients.

Longitudinal and abattoir-based surveillance of MERS-CoV in camels in Jordan, 2018-2020.

To generate baseline information to help better understand the antibody kinetics and nasal shedding dynamics of MERS-CoV in camels in Jordan, a longitudinal surveillance study was conducted in two phases; phase 1 was between December, 2018 and January, 2019 and phase 2 between August and December 2020. In each phase, two camel herds were studied. These herds were located in Al-azraq and in Al-ramtha area and were named Al-azraq and Al-ramtha herds, respectively. The same camel herd of Al-zarqa area was sampled in both phases while two different camel herds, one in each phase, were sampled in Al-ramtha area. Blood and nasal swabs were collected from same selected animals in all visits to each herd in both phases. Additionally, nasal swabs and retropharyngeal lymph node tissue samples were collected from sixty-one camels slaughtered at Al-ramtha abattoir during phase 2 to enhance virus isolation opportunities and phylogenetic analysis. All sampled animals from Al-azraq camel herd were either borderline or seropositive on spike 1 based ELISA assay and negative on quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) in both phases. In Al-ramtha camel herds, an unsteady pattern prevailed in animals' seropositivity in both phases and viral RNA was detected in all animals in the end of phase 1 and in one animal during phase 2. For the seroconversion, anti-MERS-CoV spike 1 antibodies were detected in two animals in phase 1 in the first collection only. While, in phase 2, intermittent seroconversion pattern was observed in several samples over time of collections that ended with all animals became seropositive in the last collection (after nineteen days from viral RNA detection). In addition, viral RNA was detected in nasal swabs of 3 slaughtered camels. Phylogenetic analysis of a partial fragment of spike 1 gene sequences of all MERS-CoV isolates clustered together with clade B of MERS-CoV. This cluster contains all MERS-CoV sequences obtained either from camels or human sources in the Arabian Peninsula indicating the continuous circulation of this clade also in Jordan.

Identification of risk factors associated with antimicrobial resistance in equine fecal Escherichia coli isolates.

Antimicrobial resistance is a growing global problem that will need a multinational collaborative effort to overcome this serious challenge. The aim of the study is to investigate the potential risk factors associated with the prevalence and distribution of antimicrobial-resistance genes (ARGs) of Escherichia coli isolates obtained from equine fecal samples. One hundred and eighteen horses from different geographical locations and management systems were enrolled in the study and a questionnaire containing information about each individual horse was designed and filled. The enrolled horses belonged to 2 main categories (Hospitalized horses (n = 31), and Non hospitalized horses (n = 87)). In total, 103 E. coli isolates were collected from the 118 horse fecal samples. Genes that are responsible for resistance to ß-lactams, tetracyclines, aminoglycosides, and trimethoprim were detected using uniplex and multiplex polymerase chain reaction (PCR). The prevalence of antimicrobial resistance was significantly higher in hospitalized horses compared to non-hospitalized ones (p ≤ .05), particularly against (trimethoprim, doxycycline, oxytetracycline, and amoxicillin-clavulanic acid). The most prevalent antimicrobial-resistant genes were aminoglycoside resistant genes (strA, strB, and aadA) in percentages; 89%, 85%, and 84%, respectively. Statistical analysis revealed a significant association between risk factors and occurrence of ARGs (p ≤ .05). Significant risk factors include the last treatment and history of antimicrobial administration, breed of horses, use of horses, type of diet fed for horses, practice management and history of last illness. Tetracycline-resistance gene (tetA) was 23 times more likely to be found in the Arabian and local breeds of horses compared to English and warmblood breed. TetA is also 8 times more likely to be found in horses that were fed a natural diet compared to other horses that were fed manufactured/ processed feed. In conclusion, E. coli bacterium can harbor high resistance to different classes of antimicrobials which increases the risk of potential uncontrolled transmission of the multi-drug resistant E. coli bacterium to veterinarians and horse handlers, as well as to the equine population itself.

Seroprevalence and associated risk factors of Coxiella burnetii (Q fever) in goats and sheep in northern Jordan.

This cross-sectional study aimed to study animal, farm, and within-farm seroprevalence of C. burnetii and to identify associated risk factors in goat and sheep farm in northern Jordan. Questionnaire was developed to collect information about risk factors and farms management practices. Blood samples from 730, ≥ 1-year-old females (goat n = 250; sheep n = 480) were randomly collected from 20 goat herds and 40 sheep flocks. IDEXX ELISA Kit was used to detect C. burnetii antibodies. The overall goat and sheep seroprevalence level was 32.5% (237/730) and was significantly higher in goats (43.3%, 108/250; 95% CI 37-49.6) than sheep (27%, 129/480; 95% CI 29.1-36.2) (χ2 test, p ≤ 0.001). Eighty percent (16/20) of goat herds and 60% (24/40) of sheep flocks had at least one seropositive animal (p ≥ 0.05). The average within goat herds and sheep flock seroprevalence were 36.4% (ranged: 0-91%) and 23.4% (ranged: 0-82%), respectively. Multivariate logistic regression model revealed that seroprevalence increased 1.79 times in goat herds compared with sheep flocks, 3.2 times more in farms containing ≥ 100 animals, and 1.7 times higher in farms with their animals that were ≥ 2 years of age than in farms with their animals that are < 2 years of age. In addition, seroprevalence significantly increased 1.52 times in farms loaning bucks or rams during breeding season and 1.63 times in farms containing cats on premises (p ≤ 0.05). Farm biosecurity measures are essential to prevent introduction and minimize transmission of C. burnetii infection to humans and animals.

Identification of Escherichia coli from broiler chickens in Jordan, their antimicrobial resistance, gene characterization and the associated risk factors.

BACKGROUND: Avian pathogenic Escherichia coli (APEC) is the principle cause of colibacillosis affecting poultry. The main challenge to the poultry industry is antimicrobial resistance and the emergence of multidrug resistant bacteria that threaten the safety of the food chain. Risk factors associated with emergence of antimicrobial resistance among avian pathogenic E. coli were correlated with the inappropriate use of antimicrobials along with inadequate hygienic practices, which encourages the selection pressure of antimicrobial resistant APEC. The aim of this study was to isolate, identify, serogroup and genotype APEC from broilers, assess their antibiotic resistance profile, expressed genes and the associated risk factors. RESULTS: APEC was isolated from the visceral organs of sick chickens with a prevalence of 53.4%. The most prevalent serotypes were O1, O2, O25 and O78, in percentage of 14.8, 12.6, 4.4 and 23.7%, respectively. Virulence Associated Genes; SitA, iss, iucD, iucC, astA, tsh cvi and irp2 were detected in rate of 97.4, 93.3, 75, 74, 71, 46.5, 39 and 34%, respectively and 186 (69.2%) isolates possess > 5-10 genes. The highest resistance was found against sulphamethoxazole-trimethoprim, florfenicol, amoxicillin, doxycycline and spectinomycin in percentage; 95.5, 93.7, 93.3, 92.2 and 92.2%, respectively. Sixty-eight percent of APEC isolates were found to have at least 5 out of 8 antimicrobial resistant genes. The most predominant genes were Int1 97%, tetA 78.4%, bla TEM 72.9%, Sul1 72.4%, Sul2 70.2%. Two risk factors were found to be associated with the presence of multi-drug resistant APEC in broiler chickens, with a P value ≤0.05; the use of ground water as source of drinking water and farms located in proximity to other farms. CONCLUSIONS: This study characterized the VAGs of avian pathogenic E. coli and establish their antimicrobial resistance patterns. The widespread of antimicrobial resistance of APEC isolates and detection of ARGs highlighted the need to monitor the spread of ARGs in poultry farms and the environment in Jordan. Use of ground water and closely located farms were significant risk factors associated with the presence of MDR APEC in broiler chickens in Jordan.

Genetic characterisation of Taenia multiceps cysts from ruminants in Greece.

This study was designed to genetically characterise the larval stage (coenurus) of Taenia multiceps from ruminants in Greece, utilising DNA regions within the cytochrome c oxidase subunit 1 (partial cox1) and NADH dehydrogenase 1 (pnad1) mitochondrial (mt) genes, respectively. A molecular-phylogenetic approach was used to analyse the pcox1 and pnad1 amplicons derived from genomic DNA samples from individual cysts (n=105) from cattle (n=3), goats (n=5) and sheep (n=97). Results revealed five and six distinct electrophoretic profiles for pcox1 and pnad1, respectively, using single-strand conformation polymorphism. Direct sequencing of selected amplicons representing each of these profiles defined five haplotypes each for pcox1 and pnad1, among all 105 isolates. Phylogenetic analysis of individual sequence data for each locus, including a range of well-defined reference sequences, inferred that all isolates of T. multiceps cysts from ruminants in Greece clustered with previously published sequences from different continents. The present study provides a foundation for future large-scale studies on the epidemiology of T. multiceps in ruminants as well as dogs in Greece.

Prevalence of Staphylococcus aureus in Imported Fish and Correlations between Antibiotic Resistance and Enterotoxigenicity.

A total of 156 Staphylococcus aureus isolates were obtained from 330 imported fresh fish samples from three countries. Selective media were used for the isolation of S. aureus, and the isolates were confirmed by PCR. The isolates were tested for mecA gene, antibiotic resistance, and enterotoxin genes (sea, seb, sec, sed, see, seg, seh, and sei). Most isolates carried sea, seg, and sei genes, and seg-sei was the most frequent enterotoxin profile. About 88.5% of the S. aureus exhibited resistance to at least one antibiotic. High resistance to penicillin and ampicillin; low resistance to tetracycline, erythromycin, rifampin, and clindamycin; and very low resistance to cefotaxime, amoxicillin-clavulanic acid, gentamicin, and ciprofloxacin were exhibited by S. aureus from the three countries. In addition, some antibiotic resistance exhibited a strong correlation (P ≤ 0.01) with enterotoxigenicity in S. aureus. The study concluded that the large amount of globally traded fish increases the possibility of intercontinental transmission of enterotoxigenic and multidrug-resistant S. aureus through fish and highlights the potential influence of local fish handling and processing on consumer health worldwide. The introduction of periodic training in food safety and hygiene is essential to increase fish handlers' awareness of good hygienic practices in handling fish. These findings also enrich the ongoing debate about the risk of methicillin- and multidrug-resistant S. aureus as a foodborne pathogen compared with drug-susceptible S. aureus.

Toxoplasmosis-Related Knowledge and Preventive Practices among Undergraduate Female Students in Jordan.

Foodborne toxoplasmosis is a leading cause of foodborne deaths and hospitalization worldwide. The level of exposure to Toxoplasma gondii is influenced by culture and eating habits. There is a scarcity of data about women's knowledge and perception of this disease. The aim of this study was to determine toxoplasmosis knowledge and preventive practices of young childbearing age women in Jordan. A descriptive cross-sectional study recruited a random sample of 1,390 undergraduate university female students and was stratified based on place of residency. About half of students (51.1%) reported having "ever" heard or read about toxoplasmosis, and almost all students (98.6%) had never been tested for toxoplasmosis. Overall, there was a lack of awareness about toxoplasmosis, its risk factors, symptoms, and timing of infection, and preventive practices. High percentages of females reported a high level of hygienic practices related to hand washing after gardening, changing cat litter, and handling raw meat. However, 16.7% of students reported eating raw meat, 26.5% usually eat traditional herbs, and 17.2% drink untreated spring water. This study establishes a baseline for the awareness levels about toxoplasmosis among young women in Jordan. These findings highlight the urgent need for toxoplasmosis awareness and preventive education for childbearing females. An effective education and outreach program should cover important topics concerning risk factors, high-risk foods, and preventive measures against toxoplasmosis.

Epidemiological Observations on Cryptosporidiosis in Diarrheic Goat Kids in Greece.

This study aimed at investigating the occurrence of Cryptosporidium spp. in diarrheic goat kids in Greece and the risk factors associated with cryptosporidiosis. Altogether, 292 diarrheic 4-15-day-old goat kids from 54 dairy goat herds of Northern Greece were examined. Oocysts of Cryptosporidium spp. were detected in 223 of 292 (76.4%) goat kids and the intensity of infection was scored as "high" in 142 samples, "moderate" in 45 samples, and "low" in 36 samples. Larger herds (>200 animals) had higher infection rates than smaller ones, although this difference was not statistically significant. Significantly higher infection rates were observed in herds during late kidding season (1 January to 30 April) compared to the early one (1 September to 31 December). These results suggest that cryptosporidiosis is very common in diarrheic goat kids in Greece, especially in large herds during the late parturition season.

Reduction of the abortion rate due to Toxoplasma in 3 goat herds following administration of sulfadimidine.

The efficacy of sulfadimidine (4 doses of 33 mg/kg body weight, IM, q48h) against Toxoplasma abortion was assessed in 3 dairy goat herds suffering from Toxoplasma abortions during the 4th month of gestation. This protocol was very effective for the control of Toxoplasma abortions (P < 0.01).

Réduction des taux d'avortements causés parToxoplasmadans 3 troupeaux de chèvres après l'administration de sulfadimidine. L'efficacité de la sulfadimidine (4 doses de 33 mg/kg poids corporel, IM, q48h) contre les avortements causés par Toxoplasma a été évaluée dans 3 troupeaux de chèvres laitières souffrant d'avortements causés par Toxoplasma durant le quatrième mois de gestation. Ce protocole a été très efficace pour le contrôle des avortements causés par Toxoplasma (P < 0,01).(Traduit par Isabelle Vallières).

Comparison of two techniques for diagnosis of cryptosporidiosis in diarrhoeic goat kids and lambs in Cyprus.

This study was conducted in the Larnaca area of Cyprus and included 28 goat and 15 sheep flocks suffering from neonatal diarrhoea (>20%). Faecal samples from diarrhoeic animals revealed that 25 of the 28 goat and 12 of the 15 sheep flocks were positive for Cryptosporidium. The ELISA was more accurate in the diagnosis of cryptosporidiosis compared to the Ziehl-Neelsen staining technique (P < 0.05). Flock size and the period of kidding/lambing were found to be the main risk factors implicated in the occurrence of neonatal goat kid/lamb cryptosporidiosis.

Ovine and caprine toxoplasmosis (Toxoplasma gondii) in aborted animals in Jordanian goat and sheep flocks.

Two hundred and fifty-five biological samples (106 aborted foetal tissue samples and 149 blood samples from aborted sheep and goats) were collected from 188 animals during the lambing season from September 2009 to April 2010 from the Mafraq region of Jordan. The sampled animals belonged to 93 goat and sheep flocks that had cases of abortion. A total of 169 (66.3%) biological samples were collected from sheep and 86 (33.7%) from goats. Seventy-six (29.8%) biological samples (45 blood and 31 tissue samples) were positive for Toxoplasma gondii by PCR assay. The positive samples were obtained from 43 sheep and 23 goats. The overall toxoplasma-specific prevalence rate was 35.1% (66/188). Forty flocks (43%) had at least one T. gondii PCR-positive animal. The risk factors related to flock health status and farm management that are hypothesized to be associated with T. gondii PCR positivity were also assessed using multiple logistic regressions. The presence of cats (OR = 4.74), a large flock size (OR = 2.76) and the method of disposing the aborted foetuses (OR = 3.77) were all statistically significant (P<0.05) risk factors that were positively associated with toxoplasma positivity in goat and sheep flocks.

Epidemiology of ovine brucellosis in Awassi sheep in Northern Jordan.

We used a combined cross-sectional and longitudinal design to estimate seroprevalence of Brucella antibodies in Awassi sheep and the incidence of abortion due to brucellosis during one lambing season, and to test risk factors. The Brucella organisms isolated from aborted fetuses and vaginal swabs were characterized as Brucella melitensis biotype 3. Seventy Awassi sheep flocks were selected randomly from Northern Jordan. Sixty two of the 70 flocks were used in the cross-sectional study and 8 flocks were monitored for three consecutive months to estimate the incidence of abortion. Questionnaire data and 602 serum samples were collected and analyzed. Thirty five flocks (56%) were brucellosis-seropositive by the Rose Bengal plate-agglutination test (RBT) and 28 (45%) by enzyme-linked immunosorbent assay (ELISA). The crude seroprevalence of brucellosis at the individual-animal level was 14.3% by RBT, 7.2% by ELISA and 2.2% using both tests in series. The flock-specific, animal-level abortion risk ranged between 2.5 and 50% (median=22.6%). The flock brucellosis-status was used as the outcome variable in a multivariable logistic regression. Grazing at communal pasture increased odds, but usage of disinfectants, previous vaccination for brucellosis, and tap water were protective. The animal-level incidence of abortion was 20% and the specific incidence risk of abortion due to brucellosis was 13%.