Slide Agglutination Test Research Articles

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Identification and characterization of Salmonella spp. from samples of broiler farms in selected districts of Bangladesh

Background and Aim:Salmonella spp. are an important group of pathogens responsible for human and animal diseases. This study aimed to estimate the prevalence and identify and characterize of Salmonella spp. isolated from broiler farms of Gazipur, Tangail, and Dhaka districts of Bangladesh. This study also evaluated the difference of Salmonella positivity status between two groups of farms, good practices adapted in broiler rearing at the project intervened farms, and non-project intervened traditional farms.Materials and Methods:A total of 352 samples including 128 cloacal swabs, 32 whole carcasses, 64 feed, 64 water, and 64 attendants’ hand rinses were collected through convenient sampling technique from 16 poultry food safety project of Food and Agricultural Organization of United Nations Bangladesh intervened farms and other 16 non-project intervened farms in the same location. Various cultural based techniques and biochemical methods were employed for the estimation of prevalence, isolation, and identification of Salmonella spp. which was further evaluated by polymerase chain reaction. Antimicrobial susceptibility test using disk diffusion methods and serogrouping by slide agglutination test was accomplished for additional characterization.Results:Among the samples, an overall prevalence of Salmonella spp. was 31.25% (110/352) (95% confidence interval [CI]=26.44-36.38%). However, the prevalence of Salmonella spp. was 24.43% (43/176) (95% CI=18.28-31.47) in project intervened farms and 38.07% (67/176) (95% CI=30.87-45.68%) in non-intervened farms. Among the 110 isolates, 31.82% (35/110) were fitted under serogroup B, and the rest of the isolates 75 (68.18%) under serogroup D. Of 110 isolates, 82.72%, 77.27%, 81.82%, and 79.09% were susceptible to ciprofloxacin, gentamycin, norfloxacin, and streptomycin, respectively. In addition, 81.82% and 80% isolates were resistant to erythromycin and tetracycline, respectively. Isolated Salmonella spp. presented moderate resistance to both amoxicillin and azithromycin. Alarmingly, 80.91% (89/110) isolates were shown to be multidrug-resistant Salmonella spp.Conclusion:The study has presented a significant variation of the prevalence of Salmonella spp. between project intervened and non-project intervened farms, and this indicates project intervened farms are comparatively safer than the non-intervened farms considering public health and food safety grounds. This research outcome also has highlighted a substantial proportion of poultry origin multidrug resistance Salmonella spp. is a potential source of public health hazards. In this regard, proper awareness creation and motivational activities on good agriculture practices in poultry rearing and maintaining good personal hygiene at the farmers’ level are warranted through participatory training.

Phenotypes, antibacterial-resistant profile, and virulence-associated genes of Salmonella serovars isolated from retail chicken meat in Egypt.

Aim:The present study was designed to investigate the occurrence and distribution of Salmonella serotypes in chicken meat samples, and to explore the susceptibility of the strains to antimicrobials, as well as their virulence-associated genes.Materials and Methods:Two-hundred retail chicken meat samples from different shops, as well as 25 stool specimens from retail shop workers, were included in the study. The collected samples were examined bacteriologically for the presence of salmonellae. Salmonella isolates were serotyped using a slide agglutination test for O and H antigens and were screened for the presence of five virulence genes (stn, pef, inv A , sop B , and avrA) using a uniplex polymerase chain reaction assay and for their susceptibility to 18 antimicrobial agents using the disk diffusion method.Results:Thirty-one Salmonella isolates belonging to 12 different serovars were identified. Salmonella Enteritidis and Salmonella Kentucky were the dominant serovars (22.6% each). Salmonella isolates displayed a high antibiotic resistance against erythromycin, sulfamethoxazole/trimethoprim, doxycycline, cephalexin, cefaclor, tetracycline, polymyxin B, cefuroxime, vancomycin, and streptomycin. All Salmonella isolates exhibited multidrug resistance (MDR) and demonstrated different virulence genes. The majority of Salmonella serovars (87.1%) harbored sopB gene, 54.8% carried avrA and pef genes, while all isolates carried invA and stn genes.Conclusion:The presence of virulent MDR Salmonellae in raw chicken meat could allow the possibility of transmission of these resistant serovars to humans. Therefore, strict hygienic measures should be followed on the whole poultry production chain to decrease the potential transmission of Salmonella infection from poultry meat to humans.

Seroprevalence of Anti-Toxoplasma gondii Antibodies and Associated Factors Among Pregnant Women Attending Antenatal Care at Debre Markos Referral Hospital, Northwest Ethiopia.

Background:Toxoplasma gondii is an obligate intracellular protozoan parasite that causes a zoonotic disease called toxoplasmosis. If the infection acquired during pregnancy is not detected and treated early, the parasite can be transmitted transplacentally to the fetus, resulting in congenital toxoplasmosis, which likely leads to serious consequences in the fetus. Toxoplasmosis constitutes a major public health problem particularly in low- and middle-income countries including Ethiopia. This study aimed to determine the seroprevalence and to assess the potential risk factors of toxoplasmosis among pregnant women attending antenatal care at Debre Markos Referral Hospital, Northwest Ethiopia.Methods:In this cross-sectional study, data on the sociodemographic and potential obstetric and behavioral risk factors were gathered through pretested structured questionnaires, and 3 mL of venous blood was also drawn from each of randomly selected 233 study subjects. The serum samples were separated from the blood samples and tested for anti-Toxoplasma antibody using Toxo-latex slide agglutination test. Logistic regression analysis was used to examine the association between risk factors considered and T gondii infection.Results:The overall prevalence of T gondii infection was 67.8%, indicating a high prevalence of toxoplasmosis in the study area. In multivariate analysis, keeping domestic cat at home was found to be the only explanatory variable of toxoplasmosis (adjusted odds ratio = 2.449, 95% confidence interval = 1.183-5.070, P = .016). All sociodemographic variables and most of the potential obstetric and behavioral risk factors were not statistically significant explanatory variables of T gondii infection.Conclusion:The prevalence of toxoplasmosis is high in the study area. Thus, pregnant women should be aware of the potential risk of the disease when keeping cats at home and management of their litter. Inclusion of serologic screening for T gondii infection at antenatal care is warranted for prevention of congenital toxoplasmosis.

A study to find the correlation between ABO blood group and pulmonary tuberculosis

Introduction: Studies regarding pulmonary tuberculosis (PT) and blood group correlations were reported from different parts of globe. But there was huge disparity. Hence a study was conducted to find the relation between PT and ABO Rh blood group. Materials and Methods: Individuals with signs and symptoms suggestive of PT were included in the study. Two spot sputum samples were collected, sputum smears were stained by ZN staining. Simultaneously, ABO blood grouping and Rh typing was done for all the participants by slide agglutination test. P value of <0.05 was considered statistically significant. All the smear positive cases were included in test group. Results: During the study period, total 421 participants were included in the study the smear positivity was 64 (14.6%). In the test group, 2.2% were A+, 0.45% were A-; statistically there was no significant difference; 4.3% were B+, 0.9% were B- statistically there was no significant difference. Whereas 1.1% were AB+ and 0.45% were AB-; statistically there was no significant difference and 3.6% were O+, 1.4% were O-; statistically there was no significant difference. Conclusion: Present study showed that there was no significant association between blood groups and PT. Studies with large sample size for long time is recommended.

Preparation of O-antigen from Yersinia ruckeri Serotype O1 and Use in the Slide Agglutination Test

In this study, it was aimed at early detection of yersiniosis caused by Yersinia ruckeri in rainbow trouts (Oncorhynchus mykiss) using agglutination test. O-antigen of Y. ruckeri isolated from diseased fishes was prepared. This prepared O-antigen was injected into the ear vein of the rabbit. Serum was removed from blood samples taken on 7th and 15th days after the last injection. Thus, the antibody of the Yersinia ruckeri was produced in the rabbit. Titers of the resulting antibodies were determined by microtitering in 96 well-based plates. It was used successfully in the slide agglutination tests by making necessary dilutions according to the titer ratios of the antibodies. We determined that the motile and Gr (-) bacteria isolated from diseased rainbow trout with yersiniozis symptom were Y. ruckeri according to the results of slide agglutination test without any biochemical test. The isolated bacteria were confirmed to be Y. ruckeri with the results of API 20E test kit and PCR test. The antibodies were stored at -20 and -80°C for use subsequent studies.

Meningococcal Meningitis: A Multicentric Hospital-based Study in Kathmandu, Nepal

Background: The global epidemiology of meningococcal disease varies markedly by region and over time. In Nepal, information on serogroup of meningococci is not available since the 1983 serogroup A epidemic in Kathmandu. Objective: To provide some fundamental data on the circulating serogroups of meningococci for potential meningococcal immunization programs in Nepal. Methods: This cross-sectional prospective study was conducted from January 2017 to December 2018 among 387 clinically suspected meningitis cases. Cerebrospinal fluid samples were collected by lumbar puncture technique at five referral hospitals of Kathmandu and processed by conventional cultural techniques. Neisseria meningitidis was identified by colony morphology, Gram staining and oxidase test. Serogrouping of meningococci was performed by slide agglutination test. Antibiotic susceptibility testing was done by the modified Kirby Bauer disc diffusion method. The data was entered into IBM SPSS Statistics 21 software and a p-value of &lt;0.05 was considered significant. Results: Thirty-two samples were positive by culture for a bacterial pathogen with 2.3% of meningococci. All except one meningococcal meningitis cases were aged below 15 years. All N.meningitidis isolates belonged to serogroup A and were susceptible to ceftriaxone, chloramphenicol, meropenem and minocycline; however, 22% isolates showed resistance to cotrimoxazole and 11% intermediate resistance to ciprofloxacin. Conclusion: The circulating serogroup of N. meningitidis in Kathmandu has not changed over the past 35 years. The prevalence of meningococcal meningitis in Kathmandu is low but might be underestimated due to the sole use of culture-based diagnostic methods. Detection of meningococci by alternative methods may be useful in the precise estimation of actual disease burden.

Self-made Salmonella Pullorum agglutination antigen development and its potential practical application

Pullorum disease caused by Salmonella Pullorum is one of the most important infectious diseases in the poultry industry worldwide, which leads to serious economic losses in many developing countries because of its high mortality rate in young chicks. The traditional slide agglutination test with low cost, fast reaction, and on-site detection has been widely used in the diagnosis of Pullorum disease. However, in practice, the test performance is with the disadvantages of false positive results and unstable detection results. In this paper, we developed self-made agglutination antigens prepared by local isolates in the poultry farm and compare the detection performance with commercial agglutination antigens (China Institute of Veterinary Drug Control) and Group D Salmonella ELISA kit (BioChek UK Ltd). The results of detecting 200 serum samples indicated that the consistency of commercial agglutination antigen detecting in 2 times was only 79.5%. Using the ELISA kit as the reference method, the commercial agglutination antigen detecting results of the Kappa test were only moderately consistent (0.58 ∼ 0.59). Meanwhile, positive and total coincidence rates of the self-made agglutination antigen test with more reliable repeat could reach 97.4 and 88%, respectively, and the result of Kappa test was highly consistent (0.75). The Receiver Operating Characteristic curve analysis clarified that the area under the receiver-operating-characteristic curve values of self-made and commercial agglutination antigen tests could reach 0.861 and 0.804, respectively. These results were coincident when detecting known positive serum from the infected chickens. It's worth mentioning that the visible positive reaction of self-made agglutination antigen test appeared faster and stronger than commercial antigen test. In conclusion, self-made Salmonella Pullorum agglutination antigen developed in this study was much better than commercial agglutination antigen and is expected to be a valuable tool in the diagnosis of the epidemiology of Salmonella Pullorum.

Estimating the Spatial Accessibility to Blood Group and Rhesus Type Point-of-Care Testing for Maternal Healthcare in Ghana.

Background: In Ghana, a blood group and rhesus type test is one of the essential recommended screening tests for women during antenatal care since blood transfusion is a key intervention for haemorrhage. We estimated the spatial accessibility to health facilities for blood group and type point-of-care (POC) testing in the Upper East Region (UER), Ghana. Methods: We assembled the attributes and spatial data of hospitals, clinics, and medical laboratories providing blood group and rhesus type POC testing in the UER. We also obtained the spatial data of all the 131 towns, and 94 health centres and community-based health planning and services (CHPS) compounds providing maternal healthcare in the region. We further obtained the topographical data of the region, and travel time estimated using an assumed tricycle speed of 20 km/h. We employed ArcGIS 10.5 to estimate the distance and travel time and locations with poor spatial access identified for priority improvement. Findings: In all, blood group and rhesus type POC testing was available in 18 health facilities comprising eight public hospitals and six health centres, one private hospital, and three medical laboratories used as referral points by neighbouring health centres and CHPS compounds without the service. Of the 94 health centres and CHPS compounds, 51.1% (48/94) and 66.4% (87/131) of the towns were within a 10 km range to a facility providing blood group and rhesus type testing service. The estimated mean distance to a health facility for blood group and rhesus POC testing was 8.9 ± 4.1 km, whilst the mean travel time was 17.8 ± 8.3 min. Builsa South district recorded the longest mean distance (25.6 ± 7.4 km), whilst Bongo district recorded the shortest (3.1 ± 1.9 km). The spatial autocorrelation results showed the health facilities providing blood group and rhesus type POC testing were randomly distributed in the region (Moran Index = 0.29; z-score = 1.37; p = 0.17). Conclusion: This study enabled the identification of district variations in spatial accessibility to blood group and rhesus type POC testing in the region for policy decisions. We urge the health authorities in Ghana to evaluate and implement recommended POC tests such as slide agglutination tests for blood group and rhesus type testing in resource-limited settings.

Development of a pooled antigen for use in the macroscopic slide agglutination test (MSAT) to detect Sejroe serogroup exposure in cattle

This study aimed to develop and evaluate a pooled antigen for use in the macroscopic slide agglutination test (MSAT) to detect cattle positive for the Sejroe serogroup. To this end, 193 bovine serum samples from different Pará State regions (Amazonia) were subjected to a reference microscopic agglutination test (MAT) for the serological diagnosis of leptospirosis using 11 serovars representing the Sejroe serogroup: Hardjo-prajitno; Hardjo-bovis; Sejroe; Wolffi; Guaricura (Bov.G.); Guaricura (M4/98); Ricardi; Gorgas; Recreo; Polonica and Medanensis. The three most prevalent serovars in the MAT were selected for the development of a pooled antigen for use in MSAT; subsequently, the 193 serum were assessed with the macroscopic slide agglutination test (MSAT) containing the developed antigen. The Kappa test was used to determine the general agreement between the MAT and MSAT results. As a result, of the 193 serum samples, 155 (80.3%) were reactive, and 38 (19.7%) were non-reactive in the MAT; Hardjo-prajitno, Wolffi and Medanensis were the three most prevalent serovars. Of the 193 serum samples tested in the MSAT using the developed pooled antigen, 114 were reactive (59.0%), and 79 (41.0%) were non-reactive; the Kappa coefficient was 0.52 (CI 95%, 0.40–0.63), indicating moderate agreement between the two tests. The MSAT with the pooled antigen including the most prevalent serovars detected bovines with the Sejroe serogroup exposure, mainly in animals with high titters in the MAT, and could be used to screen herds suspected of acute infection by this serogroup in Pará State.

Isolation, conventional and molecular characterization of Salmonella spp. from newly hatched broiler chicks

Salmonella is an important pathogen for poultry production as well as for human due to zoonotic importance. It has more than 2600 identified serovars despite of this identification and classification of Salmonella isolates into different serovars is critical for study of incidence and surveillance. This study investigates the epidemiology and molecular characterization of Salmonella isolates in broiler chicks during 1st week of life. A total of (n = 1000) samples including liver, intestine, yolk sac, spleen and heart blood were collected from El-Gharbia, El-Behera, Kafr-Elshikh, Alexandria, Marsamatroh Provinces in Egypt and tested through bacteriological, biochemical, serological and molecular examinations. Incidence of Salmonella was demonstrated on 75 positive samples from 1000 samples and the predominance of Salmonella that isolated from internal organs of newly hatched chicks was highest from yolk sacs (10%), liver and intestines (9%) followed by the spleen (7.5%) then heart blood (2%). Serotyping of the isolated strains using slide agglutination test revealed that 24 isolates belonging to S. enteritidis (1,9,12 g.m 1,7), while, 14 isolates belonging to S. virchow (6,7 r 1,2), in addition to, 12 isolates belonging to S. typhimurium (1,4,5,12.i.1,2) and 8 isolates belonging to S. kentucky (6,8.I,z). Enterobacterial Repetitive Intergenic Consensus (ERIC) PCR revealed that two S. enteriditis isolates were identical and one isolate differ by 40%, while two S. typhimurium isolates were identical by 80% and one isolate was similar by 20% to the other two isolates, in addition, two S. virchow isolates were identical by 80% and the two S. kentucky isolates were different.

A promising detection candidate for flagellated Salmonella spp.

Salmonella is a common and important pathogen for both human and animals. All Salmonella except Salmonella pullorum and Salmonella gallinarum have flagellum. Flagellin (FliC) is the main subunit protein forming the bacterial filament, which is present in large amounts on the surface of all flagellated Salmonella. After bioinformatics analysis, the most highly conserved region (locates position from 1 to 102 amino acid residue of FliC, we named it as FliC′) was selected, and corresponding recombinant FliC′ (rFliC′) protein was tailored as an immunogen to generate monoclonal antibodies (MAbs) against Salmonella flagellin. BALB/c mice were immunized with the purified recombinant protein rFliC′, which were prepared by prokaryotic expression system pET22b (+) expressing FliC′. After fusion of spleen cells from the immunized mice and SP2/0 cells, three hybridoma cells (1D6, 2G6 and 3E2) producing MAbs against targeted flagellate Salmonella FliC′ were generated and screened. The ability of MAb 3E2 to recognize and bind to Salmonella flagella was demonstrated by immunogold electron microscopy (IEM) method. Western blot (WB) analysis demonstrated that MAb 3E2 could specifically recognize flagellated Salmonella strains. Moreover, MAb 3E2 has a direct agglutination activity against Salmonella strains with visible agglutination reaction. To further verify this agglutination activity, a total of 52 flagellated Salmonella strains (23 serovars), 8 non-flagellate Salmonella strains (2 serovars) and 16 other non-Salmonella bacteria strains were used to evaluate the specificity of the MAb by direct Slide Agglutination Test (SAT). Results showed that MAb 3E2 reacted with all Salmonella strains possessing flagellum and had no cross-reaction with non-flagellate Salmonella strains or other non-Salmonella bacteria strains. Sequentially, the ability to detect the presence of Salmonella in raw samples of the MAb 3E2-based SAT method was evaluated. The conventional culture-based detection method was performed as the standard reference method for detection of Salmonella. Altogether, 369 samples collected from laying hens were tested, and the results indicated that the MAb 3E2-based SAT method could specifically detect Salmonella. Furthermore, the SAT results were obtained more quickly, as compared with the standard method. As a whole, the MAbs against the tailored conserved region of Salmonella flagellin were prepared in this study, and MAb 3E2-based SAT is a promising candidate for the flagellated Salmonella spp. rapid detection.

Antimicrobial Resistance Profile of Salmonella Typhimurium Isolated from Commercial Poultry and Poultry Farm Handlers in Nasarawa State, Nigeria

Aim: This study was designed to investigate the antimicrobial resistance profile of Salmonella Typhimurium isolated from commercial poultry and handlers in Nasarawa State, Nigeria.&#x0D; Study Design: Investigative.&#x0D; Place and Duration of Study: Microbiology Laboratory, Nasarawa State University, Keffi, Nigeria, from 1st November 2017 to 31th April 2018.&#x0D; Methodology: A total of 1500 samples (poultry droppings, poultry flesh, feeds, handlers’ faeces and hand swabs) were screened for the presence of Salmonella Typhimurium using pre-enrichment and selective enrichment culture media. Subculture of inoculated samples was done on Salmonella-Shigella agar and Xylose Lysine Deoxycholate agar. Presumptive Salmonella colonies were confirmedas serovar Typhimurium using both the conventional biochemical screening tests and Microgen Bio product GN identification system and slide agglutination test using polyvalent antisera. Antimicrobial susceptibility testing and interpretation were carried out as described by the Clinical Laboratory and Standards Institute guidelines.&#x0D; Results: Resistance was highest to Augmentin (98.1%) and lowest to Imipenem (1.0%). No resistance was observed in all the isolates from poultry handlers to Ceftriaxone, Ceftazidime, Gentamicin and Streptomycin; but all were resistant to Ampicillin and Augmentin. The resistance of isolates from poultry and handlers to all the antibiotics is significant (χ 2 = 13.037; P = 0.01). Most (86.7%, 92/106) resistant isolates belong to the multiple drug resistance class.The distribution of classes of resistance of isolates from poultry and handlers is significant (χ2 = 318; P = 0.00). MARI is greater than 0.2.&#x0D; Conclusion: Salmonella Typhimurium with increasing multidrug resistance to antibiotics especially the β-lactam antibiotics has emerged in poultry.

Conventional and molecular detection of vibrio cholerae isolated from environmental water with the prevalence of antibiotic resistance mechanisms.

Environmental water is an important source for Vibrio cholerae, which is autochthonous to the aquatic environment, monitoring this bacterium in water is important for control of cholera. Vibrio cholerae represents an enormous public health problem around the world, especially in developing countries. One hundred samples were collected and selected. The samples were filtered and transferred to slants containing 2ml of alkaline peptone water, then subcultured on selective medium Thiosulphate Citrate Bile Salt Sucrose agar. All presumptive isolates were confirmed by using a series of biochemical tests including Oxidase test, Simmon Citrate test, DNase test, Indole test, Klingler Iron Agar (KIA) test, MacConkey agar test and motility. Confirmed Vibrio cholera strains were then screening for slide agglutination test by using commercially antisera polyvalent and monovalent O1 and O139 for determining strain serotype. The DNA extracted from pure culture and Polymerase Chain Reaction assay was used for molecular detection of Vibrio cholerae, a specific primer which designed according to ctxA gene sequences. This primer was detection and amplifying 241 base pairs of the ctxA gene. The resistance to antibiotics by Vibrio cholerae was determining by using thirteen standardized disc diffusion including Amikacin, Ceftriaxone, Ceftazidime, Gentamycin, Tetracycline, Streptomycin, Tobramycin, Cephotaxime, Nalidixic Acid, Norfloxacin, Cephalothin, Rifampicin, Cefixime. From one hundred water samples were detected, fifty-six samples were motile and positive for biochemical tests. Fifteen isolates confirmed as Vibrio cholera by Polymerase Chain Reaction (PCR) assay with primers de­signed for ctxA and 241bp band was observed. These fifteen isolates showed agglutination with polyvalent and monovalent O1 antisera, and two strains represented Ogawa from other strains that showed Inaba. The fifteen isolates exhibited an identical response to each antibiotic examined. They showed sensitive to all antibiotics except Amikacin, Streptomycin, Cefixime, Norfloxacin, Cephalothin. the aim of this study was determined the accurate method for detection of Vibrio cholerae in environmental water. In the current study, we found that the molecular method using Polymerase Chain Reaction performance using the ctxA gene-specific primers for detection of Vibrio cholerae was faster and accurate and specific.

Isolation and identification of pathogenic bacteria from ready-to-eat fast foods in Al-Quwayiyah, Kingdom of Saudi Arabia.

Food-borne pathogens are becoming a globally formidable health problem and perceived as a major health concern in the Kingdom of Saudi Arabia (KSA). Contamination ensued through unclean raw food materials and particles, use of polluted water, unhygienic preparation processes and use of contaminated containers. Herein, the prevalence of food-borne pathogens in ready-to-eat (RTE) fast foods from fifteen different food eateries such as 7 restaurants, 6 cafeterias and 2 two college canteens in Al-Quwayiyah, Riyadh Region of Saudi Arabia was studied. Microbiological analysis of 155 fast food samples which included, Vegetable salad, Falafel, Kibtha and Shawarma. The isolates were detected using biochemical tests and API 20E and slide agglutination test were conducted for Salmonella spp. detection. Bacterial growth was found in all food samples tested. Moreover, the test also showed high levels of total aerobic count: vegetable salad 6.34+0.03, falafel 5.79 + 0.18, kibtha 5.06 +0.02 and shawarma 3.54 + 0.13. Organisms isolated include Salmonella spp. (15%), Escherichia coli (18%) and Staphylococcus aureus (7%). Salmonella is one of the most virulent pathogen implicated in food-borne disease outbreaks. There are numerous transmission routes for Salmonellosis , but the majority of the human infections are derived from consumption of contaminated poultry products. Consistently, Salmonella Enteritidis, Salmonella Typhimurium and Salmonella Heidelberg are the three most frequent serotypes recovered from humans each year. Serologically identified Salmonella serotypes from RTE fast food samples were Salmonella Typhimurium with 65%, the most predominant one compared to Salmonella Enteritidis that was 35%. The bacterial count of vegetable salad, falahfel, kibtha was statistically significant when compared with Shawarma ( p < 0.05). This result indicated that most of the ready-to-eat food samples examined in the study did not meet any bacteriological quality standard as recommended by The New South Wales (NSW) Food Authority to be <5.0 log10 CFU g−1 and, therefore, it poses potential risks to consumers. Ready- to- eat fast foods must be cooked and served to the consumers with all hygienic measures. Keywords: foodborne pathogen, microbial quality, Ready- to- eat fast foods, Al-Quwayiyah Afr. J. Food Agric. Nutr. Dev . 2019; 19(3): 14739-14751

Seroprevalence of Brucella canis in dogs and at-risk humans in Jordan

Brucella canis infection is a neglected zoonotic disease and its seroprevalence in dogs and at-risk humans has not been previously studied in several countries including Jordan. The main aim of this study was to determine the seroprevalence and identify risk factors of B. canis infection in police, breeding and stray dogs and in at-risk humans in Jordan. A total of 169 sera samples from apparently healthy dogs and 185 samples from apparently healthy people (85 from dog handlers and 100 from the general population) were tested in the study. Antibodies against B. canis were tested using the canine D-Tec&lt;sup&gt;®&lt;/sup&gt; CB Rapid Slide Agglutination Test (RSAT) kit with secondary 2-mercaptoethanol (ME-RSAT). Overall, 8.3% of the dog sera samples tested positive to antibodies against B. canis, and 37.8% of stray dogs tested positive. Seroprevalence was higher in male dogs than in females. Furthermore, none of the tested human samples was positive to antibodies against B. canis. There was a significant association between seropositivity and the type of dog. The study reports preliminary findings that suggest the presence of B. canis among stray dogs in Jordan. Thus, preventive measures should be taken to control the transmission of this pathogen from stray dogs to other dogs and humans as well.

Detection of shiga toxin strains of Escherichia coli non O157 in different soft cheese by polymerase chain reaction

The objectives of the current study were detection and characterization of the isolated Escherichia coli non O157 using PCR assay. A total of 90 cheese samples high salt soft cheese, Kareish cheese and Tallaga cheese were examined for E. coli non O157 using modified vancomycin- trypticase soy broth and Sorbitol MacConkey agar plates. Serodiagnosis of E. coli non-O157 has been done using slide agglutination test. Toxigenic E. coli non-O157 isolates were detected using PCR assay. Results postulated that the detection rate of non O157 using biochemical technique was 63.33% in Kariesh cheese, 20% in high salt soft cheese cheese, and 33.33% in Tallaga cheese. Ten different serotypes of E. coli non O157 have been distributed as following O1 (4.44%), O18 (1.11%), O20 (1.11%), O25 (1.11%), O26 (3.33%), O125 (1.11%), O126 (1.11%), O127 (1.11%) and untyped E. coli (18.89%). Regarding PCR results, serotypes group (O1) was positive for Stx2 gene, (O1 and O20) were positive for both Stx2 and hly genes, one serotype group O25 was positive for eaeA gene, one serotype group O26 was positive for hly gene while no detection for Stx1and STa genes. From the aforementioned data, attention must be paid to the problems of E. coli non O157 in foods. Consequently, more restriction and preventive measures should be taken in milk herds, milk production and dairy factories in respect to quality control sanitation and health care.

Seroprevalence of Brucella canis antibodies in dogs entering a Minnesota humane society, Minnesota, 2016–2017

BackgroundCanine brucellosis, caused by the bacterium Brucella canis, is a zoonotic and largely reproductive disease of dogs. The disease is a recognized problem in canine breeding populations, and the risk to individuals assisting with birthing is well described. Prior to 2015, all cases of canine brucellosis reported to the Minnesota Board of Animal Health were in dogs used for breeding. In 2015, canine brucellosis was identified in eight Minnesota rescue dogs, all originating from specific geographic areas in South Dakota. Our objective was to measure the seroprevalence of B. canis in stray and previously owned dogs entering a large Minnesota animal rescue organization to determine if our observations represented a localized or generalized disease issue among rescue dogs. MethodsA stratified random sample of stray and previously owned dogs entering the largest Minnesota animal rescue organization between November 1, 2016 and November 7, 2017, was tested for B. canis antibodies by the 2-Mercaptoethanol Rapid Slide Agglutination Test (2ME-RSAT) (Zoetis d-TEC® CB kit). Sample sizes for each strata were calculated using previously published seroprevalence estimates. Blood from selected dogs was collected, serum harvested, and transported to the Minnesota Veterinary Diagnostic Laboratory for testing. Positive samples in the 2ME-RSAT were shipped to Cornell University for confirmation by Agarose Gel Immunodiffusion (AGID) testing. Demographics, state and setting of origin, and health status were collected on study-dogs. ResultsOf the 10,654 dogs accepted by AHS during the study period, 943 (8.9%) were selected for testing. Most study dogs arrived from Oklahoma (28%), Alabama (18%), and Minnesota (12%). The median age of study dogs was 1.5 years; 303 (32%) were intact males and 294 (31%) were intact females. Most study dogs were strays (n = 716, 76%). Of the total, 22 (3.1%) stray and eight (3.5%) owner-surrendered dogs were presumptively positive by RSAT; one (0.11%) of the stray dogs was positive by 2ME-RSAT and confirmed by AGID. The positive dog was a healthy-appearing 1 year-old neutered male beagle from Texas. ConclusionsThe seroprevalence of canine brucellosis in dogs entering Minnesota for adoption from multiple states was low. Never-the-less, care must to be taken to consider all potential risks and outcomes of interstate and international dog trade, including the spread of infectious diseases such as canine brucellosis.

Molecular epidemiology and antimicrobial resistance of diarrheagic Escherichia coli among children with acute diarrhea

Objective To learn the distribution, epidemiology and antimicrobial susceptibility of diarrheagenic Escherichia coli (DEC) isolated from patients with acute diarrhea among children less than 5 years old. Methods Totally 684 stool samples collected from Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology between August 1, 2015 and September 30, 2016 were tested by culture and identified the common pathogens. PCR was applied to detect the virulence genes of DEC. Meanwhile, serotyping of enteropathogenic E. coli (EPEC) was performed by slide agglutination tests for all the isolates of EPEC. An antimicrobial sensitivity test was performed using the agar dilution method. Results A total of 149 (21.7%) enteric bacteria pathogens were isolated from 684 specimens. DEC was found in 54 cases, ranked 2nd among the pathogenic bacteria. DEC tended to occur in summer/autumn periods. EPEC was the most frequent DEC genotype, accounted for 50% (27/54). Among EPEC, atypical EPEC was dominant, accounted for 77.8% (21/27) and typical EPEC only accounted for 22.2% (6/27). Followed by enteroaggregative E.coli 20.4% (11/54), enterotoxigenic E.coli 14.8% (8/54), enteroinvasive E. coli and Shiga toxin-producing E.coli 3.7% (2/54), 7.4% (4/54) cases were co-infected with more than one DEC genotypes. About 17/18 of suspicious DEC isolates can get the same genotypes by commercial multiplex PCR kit and single PCR test. Among the 27 EPEC strains, only 11(40.7%) strains can be detected by the slide agglutination serotyping method. More than 50% (27/54) of DEC isolates were resistant to conventional first-line antibiotics (ampicillin, trimethoprim-sulfamethoxazole) and cefazolin, cefuroxime, cefotaxime, but relatively low resistance to cefoxitin, amikacin, piperacillin/tazobactam, imipenem and meropenem. However, there was still a 9.2% (5/54) resistance rate to carbapenems. Conclusions DEC strains exhibited a high frequency of resistance to many antibiotics. Empirical antimicrobial therapy of severe DEC infection faces the challenge from the high resistance to ampicillin, trimethoprim-sulfamethoxazole. Even worse, some strains were resistant to relatively efficient drugs imipenem and meropenem. It is necessary to strengthen the epidemiological survey and antimicrobial resistance of DEC. Key words: Diarrhea; Escherichia coli; Virulence; Escherichia coli infections; Drug resistance, bacterial; Epidemiologic studies

Development of a selective medium and antisera for Pseudomonas syringae pv. syringae from seeds of barley and wheat

Bacterial black node, a disease of barley and wheat, is caused by Pseudomonas syringae pv. syringae (synonym pv. japonica, hereafter Psj). Here a selective medium, serine-potassium tellurite-based Psj-selective agar (SPTPsjA) was developed to isolate and detect Psj. After 5–7 days on SPTPsjA at 25 °C, characteristic black Psj colonies formed with an efficiency equal to that on potato–peptone–glucose agar. Except for P. viridiflava, P. cichorii, and some strains of P. syringae group bacteria, SPTPsjA either inhibited growth of other phytopathogenic bacteria or enabled their discrimination. To facilitate diagnosis of Psj, we prepared three antisera against three strains; in a slide agglutination test, the antisera reacted to all tested strains of Psj and failed to react to P. viridiflava and P. cichorii although some strains of other P. syringae group bacteria were indistinguishable from Psj with SPTPsjA and antisera. When barley and wheat seeds from fields in which bacterial black node had occurred were placed on SPTPsjA in the dark at 25 °C for 7 days, black colonies grew on the medium around the seeds. Among these strains, only those responsive to the above antisera were pathogenic on barley. These results show that the combination of SPTPsjA and antisera can isolate Psj from seeds of barley and wheat.