Avian Polyomavirus Among Psittacine Birds in Iran: Molecular Detection Rate and Associated Risk Factors.

Avian polyomavirus (APV) infection causes various health problems in psittacine species, including death. The present study was conducted to investigate the prevalence of APV among psittacine birds in Iran. We also aimed to evaluate the impact of age, sex, species, season, and origin of the birds on the prevalence of APV. This study investigated the presence of APV among 1050 individual birds from 7 psittacine species over a 1-year period in Iran, namely, green-cheeked parakeets (Pyrrhura molinae), rosy-faced lovebirds (Agapornis roseicollis), monk parakeets (Myiopsitta monachus), sun conures (Aratinga solstitialis), Senegal parrots (Poicephalus senegalus), cockatiels (Nymphicus hollandicus), and grey parrots (Psittacus erithacus). The overall prevalence of APV in all studied species was 25% (263/1050, 95% confidence interval [CI]: 22.5-27.8). Results of the study showed that age and the season of the year were 2 important determinant factors in the prevalence of APV in psittacine birds. Young psittacine birds <6 months old were 2.94 (95% CI: 1.19-7.27) times more likely to be infected with APV than birds >1 year old, and there was a significant interaction between season and species in the multivariate analysis. In the winter season, rosy-faced lovebirds and green-cheeked parakeets were 15.6 (95% CI: 4.20-57.95) and 4.76 (95% CI: 1.4-16.21) times more likely to be infected with APV than in other seasons, respectively. This is the first report on the detection rate of APV in psittacine birds in Iran.

Study on antibiotic resistance and phylogenetic comparison of avian-pathogenic Escherichia coli (APEC) and uropathogenic Escherichia coli (UPEC) isolates.

Avian pathogenic Escherichia coli (APEC) and uropathogenic E. coli (UPEC) can cause vast infections in humans and poultry. The present study was conducted to compare the isolates of the APEC and UPEC pathotypes on the basis phenotypic and genotypic features of antibiotic resistance and phylogenetic differences. Total number of 70 identified E. coli strains, including 35 APEC and 35 UPEC isolates, were isolated from avian colibacillosis and human urinary tract infection (UTI), and were subjected to the antimicrobial susceptibility testing, polymerase chain reaction (PCR) detection of the resistance genes, phylogenetic grouping and DNA fingerprinting with enterobacterial repetitive intergenic consensus PCR (ERIC - PCR) to survey the variability of the isolates. The most resistance rates among all E. coli isolates were, respectively, obtained for Ampicillin (84.20%) and sulfamethoxazole-trimethoprim (65.70%). The APEC and UPEC isolates showed the most susceptibility to imipenem and gentamycin, respectively. Among 70 APEC and UPEC isolates 34.20%, 32.80%, 20.00%, and 12.80% belonged to the A, B2, D, and B1 phylogenetic groups, respectively. Analysis of the DNA fingerprinting phylogenetic tree showed 10 specific clusters of APEC and UPEC isolates. According to the results, the most effective antibiotics and the phenotypic and genotypic predominant resistance patterns of the APEC and UPEC isolates were different. Moreover, APECs and UPECs showed various dominant phylogenetic groups. With all descriptions, the APEC isolates still are potential candidates for carrying important resistance genes and can be one of the possible strains related to human infections.

Antibiotic resistance and phylogenetic comparison of human, pet animals and raw milk Staphylococcus aureus isolates.

The present study was conducted to compare the S. aureus isolates from different sources in the basis of resistance phenotypic and genotypic features and phylogenetic differences. Total of 70 S. aureus isolates (including 25 human, 25 raw milk and 20 pet animal isolates) were subjected to the antimicrobial susceptibility testing, polymerase chain reaction (PCR) detection of the resistance genes and DNA fingerprinting using random amplification of polymorphic DNA-PCR (RAPD-PCR) to survey the variability of the isolates. Among 70 S. aureus, 55 (78.5%) isolates were MRSA. The isolates showed the highest antibiotic resistance to methicillin, ampicillin and penicillin (78.5%) and showed the lowest resistance to ciprofloxacin (12.8%). ErmB and tetM resistance genes were present in all isolates and the vanA gene was not detected in any of the isolates. Thirteen distinct clusters were identified in RAPD-PCR fingerprinting. Statistical analysis showed that the isolates without resistance to antibiotics were significantly in associated with raw milk origin (P < 0.05). According to the results of the study, S. aureus strains with pets and raw milk origin are significant sources of antibiotic-resistant isolates such as MRSA. They are also carriers of resistance genes that can be transmit to human isolates and cause drug resistance in human infections. Identifying the source of these infections is possible with a reliable genotyping method such as RAPD-PCR.

Antimicrobial resistances, and molecular typing of Campylobacter jejuni isolates, separated from food-producing animals and diarrhea patients in Iran.

The aims of this study were to regain new epidemiology information about frequency, drug resistance rates, and typing of Campylobacter jejuni (C. jejuni) isolates, obtained from some poultry and cattle farms, slaughterhouses, and people with diarrhea. In this regard, Minimal Inhibitory Concentration (MIC) of several antibiotics and the associated antibiotic resistance genes, including tetO, tetA, cmeB, and blaOXA-61 were evaluated. The isolates were also typed, using the Fla-RFLP method. Generally, between 233 food animal samples, 80 (34.33%) C. jejuni were isolated. Moreover, 20 out of 74 (27%) human specimens suspected to infectious diarrhea were C. jejuni positive. High frequencies of resistance to tetracycline (100%), ciprofloxacin (95%), and nalidixic acid (86%), and low frequencies of resistance to florfenicol (0%), erythromycin (5%), and gentamicin (8%) were observed. Furthermore, in the tetracycline-resistant isolates, the existences of tetO, tetA, and cmeB were 86%, 23%, and 48%, respectively. There was a significant correlation between the cluster types obtained from Fla-RFLP method and antibiotic resistance pattern. The results suggested that the genomic link between Campylobacter spp. should be always evaluated in each country to provide an insight about the Campylobacter spp., spread in the region, in order to implement the health-controlling programs efficiently.

Prevalence and antibiotic resistance pattern of bacteria isolated from urinary tract infections in Northern Iran.

BACKGROUND: This study aimed to investigate the bacteria associated with urinary tract infection (UTI) and antibiotic susceptibility pattern of the isolates during 2013-2015 in Northern Iran. MATERIALS AND METHODS: Overall 3798 patients with clinical symptoms of UTI were subjected as samples, and they were cultured and pure isolated bacteria were identified using biochemical tests and subjected to antibiogram assessment using disc diffusion method. RESULTS: Totally, 568 (14.96%) from 3798 patients had positive UTI. Four hundred and ninety-seven (87.5%) from 568 isolated bacteria were resistant to at least one antibiotic. Escherichia coli, Staphylococcus spp., and Pseudomonas spp. were the most prevalent bacteria. Isolated bacteria indicated the highest antibiotic resistance to methicillin (76.06%) and ampicillin (89.29%) and also revealed the most sensitivity to imipenem (99.1%) and amikacin (91.57%). Statistical analysis of the resistance pattern trend during 3 years indicated the insignificant increase (P > 0.05) in antibiotic resistance of the isolates. CONCLUSION: The results of this study revealed a great concern for emerging UTI-related multidrug-resistant strains of bacteria causing UTI in Iran.

Antimicrobial Resistance and Virulence-Associated Genes of Campylobacter spp. Isolated from Raw Milk, Fish, Poultry, and Red Meat.

This study was designed and conducted to evaluate the frequency, antimicrobial resistance, and presence of six virulence-associated genes among thermophilic Campylobacters isolated from raw milk, poultry (chicken, turkey, and duck), fish, cattle, and sheep meat. Out of 590 samples, which were recovered from different origins, 141 (23.9%) samples were positive for Campylobacters. Campylobacter spp. was isolated in 40.8% (106/260), 14% (28/200), and 8.7% (7/80) of poultry meat, red meat, and milk samples, respectively. Antimicrobial susceptibility test indicated a high frequency of resistance to ciprofloxacin, tetracycline, and nalidixic acid among the isolates. Furthermore, prevalence of waaC, ciaB, and pldA genes were 91.7%, 86.7%, and 80.8%, respectively; and, none of the isolates harbored both wlaN and cgtB genes, simultaneously. Moreover, there was a weak correlation between antibiotics resistance and presence of the pathogen genes. However, the existence of Campylobacter spp. isolates in food animal products, with high resistance to antibiotics and several virulence gene possessions, is alarming and increases the attention to the widespread use of antibiotics.

Prevalence and antibiotic resistance profile of thermophilic Campylobacter spp. of slaughtered cattle and sheep in Shiraz, Iran

Although poultry meat is considered as the main source for human Campylobacter infections, there is limited information about non-poultry sources. The present study was aimed to investigate the prevalence and the antibiotic resistance of thermophilic Campylobacter spp. in fecal samples of the cattle and sheep in Shiraz, Iran. A total of 302 fecal samples were obtained from clinically healthy, slaughtered cattle and sheep from Shiraz slaughterhouse. The animals were clinically healthy before being slaughtered. The samples were cultured according to the specific cultivation method under thermophilic conditions. The susceptibility of Campylobacter isolates were determined for 13 antimicrobial agents. All enriched samples and cultured isolates were targeted for polymerase chain reaction (PCR) detection of 16S rRNA and multiplex PCR for determining their species. Among 302 fecal samples, 65 (21.5%) and 205 (67.8%) samples were positive for the presence of Campylobacter species with the cultivation and PCR techniques, respectively. All 65 distinct isolates were susceptible to neomycin and colistin and the isolates showed high resistance to cephalotin (83.0%) and ciprofloxacin (67.7%). After the multiplex PCR, 78.5% of total positive samples showed the simultaneous presence of Campylobacter jejuni and Campylobacter coli. In conclusion, the results emphasized that non-poultry farms are important as a possible source of Campylobacter infections.

Inhibitory effect of Zataria multiflora Boiss. essential oil, alone and in combination with monolaurin, on Listeria monocytogenes.

Listeria monocytogenes is one of the major causes of infections in developing countries. In this study, chemical composition and anti-listerial effect of the essential oil of Zataria multiflora Boiss. alone and in combination with monolaurin were evaluated at different pH values (5, 6, and 7) and temperatures (5 ˚C and 30 ˚C). Chemical composition of Zataria multiflora Boiss. essential oil was evaluated by gas chromatography-mass spectrometry (GC-MS) analysis. Minimum inhibitory concentration (MIC) of the essential oil and monolaurin were determined using microbroth dilution method and the interactions of essential oil and monolaurin were determined by the evaluation of fractional inhibitory concentrations (FIC) index. Carvacrol (63.20%) and thymol (15.10%) were found as the main components of the essential oil. The MIC values of the oil and monolaurin at pH 7 and 30 ˚C were measured as 312.50 µg mL(-1) and 125.00 µg mL(-1), respectively. Combination of monolaurin and Z. multiflora essential oil were found to act synergistically (FIC index < 0.5) against L. monocytogenes under different pH and temperature conditions. Decrease in the pH and temperature values have increased the anti-listerial activity of monolaurin and the essential oil. The lowest MIC value of monolaurin and essential oil was observed at pH 5 and 5 ˚C. According to our results, the oil alone or in combination with monolaurin at low pH and temperature conditions showed a promising inhibitory effect on L. monocytogenes.

Occurrence and antimicrobial resistance of emergent Arcobacter spp. isolated from cattle and sheep in Iran.

This study is conducted to determine the occurrence and antimicrobial resistance of Arcobacter spp. isolated from clinically healthy food animals. A total of 308 samples from cattle (200) and sheep (108) were collected from Shiraz slaughterhouse, southern Iran to investigate the presence of the important Arcobacter spp. using cultivation and Polymerase Chain Reaction (PCR) methods. Antimicrobial susceptibility of Arcobacter isolates was determined for 18 antibiotics using disk diffusion method. Among 308 samples, 27 (8.7%) and 44 (14.28%) were positive for the presence of Arcobacter species with cultivation and PCR procedures, respectively. The predominant species was A. butzleri in both cattle (58.33%) and sheep (55%). In addition, concurrent incidence of the species was observed in 25% of the positive samples. All Arcobacter isolates were resistant to rifampicin, vancomycin, ceftriaxone, trimethoprim and cephalothin. The isolates showed high susceptibility to tetracycline, oxytetracycline, erythromycin, ciprofloxacin, kanamycin, amikacin, gentamicin and enrofloxacin. No significant difference among cattle and sheep isolates in resistance pattern was observed. The results indicate that cattle and sheep are significant intestinal carriers for Arcobacter spp. Moreover, tetracycline and aminoglycosides showed great effects on Arcobacter species in antibiogram test and can be used for treatment of human Arcobacter infections.

Genetic Characterization of Campylobacter Jejuni and C. coli Isolated From Broilers Using flaA PCR-Restriction Fragment Length Polymorphism Method in Shiraz, Southern Iran.

BACKGROUND: Thermophilic campylobacters, particularly Campylobacter jejuni and C. coli are the main agents of human campylobacteriosis. Campylobacter contaminated chicken products is the most important source of foodborne gastroenteritis. Evaluation of genetic diversity among Campylobacter population is critical for understanding the epidemiology of this bacterium and developing effective control strategies against Campylobacter infections and other related disorders. OBJECTIVES: The aim of this study was to investigate the polymorphism of thermophilic Campylobacter isolated from broiler fecal samples in Shiraz, southern Iran. MATERIALS AND METHODS: Ninety Campylobacter isolates were recovered from broiler feces using enrichment process followed by cultivation method. The isolates were species typing on the basis of polymerase chain reaction (PCR) detection of 16SrRNA and multiplex PCR for determining two thermophilic species. To evaluate strain diversity of thermophilic Campylobacter isolates, flaA PCR-Restriction Fragment Length Polymorphism (RFLP) was performed using DdeI restriction enzyme. RESULTS: All 90 Campylobacter isolates confirmed by m-PCR were successfully typed using flaA-PCR-RFLP. Eleven different types were defined according to flaA-typing method and the RFLP patterns were located at three separate clusters in RFLP image analysis dendrogram. CONCLUSIONS: Campylobacter jejuni isolates significantly showed more variety than C. coli isolates. A relatively low genetic diversity existed among C. jejuni and C. coli isolated from broilers in Shiraz, southern Iran. In our knowledge, this was the first report of genetic diversity among broiler originated human pathogen thermophilic campylobacters in Shiraz, southern Iran.

Tetracycline Resistance Genes in Campylobacter jejuni and C. coli Isolated From Poultry Carcasses.

BACKGROUND: Campylobacter is one of the leading bacterial species causing foodborne illnesses in humans. Antimicrobial agents have been extensively used for treatment of Campylobacter infections; but in the recent years, both animal and human isolates of this bacterium have shown resistance to several antibiotics such as tetracycline. OBJECTIVES: The aim of this study was to investigate the presence of genetic determinants of tetracycline resistance in Campylobacter spp. recovered from poultry carcasses in Shiraz, Iran. MATERIALS AND METHODS: Eighty-three thermophilic Campylobacter spp. Isolates were first identified based on multiplex polymerase chain reaction (PCR) and then screened for presence of tetracycline resistance genes (tet (A), tet (B), tet (O) and te (S)) by PCR. RESULTS: The overall prevalence of Campylobacter jejuni and C. coli among the examined isolates was 51.8% and 48.2%, respectively. Tetracycline resistance genes of tet (B) and tet (S) were not seen among these Campylobacter spp. Isolates, whereas the most common tet gene identified was tet (O), found in 83.1% (69/83) of all the isolates. The tet (O) gene sequence comparison between C. jejuni and C. coli showed 100% similarity and these sequences (JX853721and JX853722) were also identical to the homologous sequences of other strains of Campylobacter spp. existing in the GenBank databases. In addition, tet (A) was found in 18% (15/83) of Campylobacter spp. isolates. To our knowledge, this represents the first report of tet (A) in Campylobacter spp. There was 100% homology between the sequences of tet (A) from this study (JX891463 and JX891464) and the tet (A) sequences mentioned for other bacteria in the GenBank databases. CONCLUSIONS: The high prevalence of tet (O) resistance gene along with new detection of tet (A) resistance gene in Campylobacter spp. isolated from poultry carcasses revealed an extensive tetracycline resistance among Campylobacter isolates from poultry in Iran. It emphasized the need for cautious use of tetracycline in poultry production to decrease the extension of tetracycline-resistant Campylobacter spp.

Correlation between distribution of muscle weakness, electrophysiological findings and CTG expansion in myotonic dystrophy.

Myotonic dystrophy type 1 (DM-1) is a multi-system disorder affecting the muscles, brain, cardiovascular system, endocrine system, eyes and skin. Diagnosis is made by clinical, electrodiagnostic and genetic studies. This study aimed to determine the correlation between CTG expansion and distribution of muscle weakness and clinical and electrophysiological findings. Genetically confirmed DM-1 patients presenting to Shariati Hospital between 2005 and 2011 were included in this study. Clinical, electrodiagnostic and genetic testing was performed and the correlation between CTG expansion and distribution of muscle weakness and clinical and electromyographic findings was studied. Thirty-three genetically confirmed DM-1 patients were enrolled. Myotonia, bifacial weakness and distal upper limb weakness were seen in all patients. Diabetes mellitus was found in one patient (3%), cardiac disturbance in eight (24.2%), cataracts in eight (24.2%), hypogonadism in five (15.2%), frontal baldness in 13 (39.4%), temporalis wasting in 14 (42.4%), temporomandibular joint disorder in seven (21.2%) and mental retardation in eight (24.2%). The mean number of CTG repeats, measured by Southern blot, was 8780 (range 500-15,833). A negative correlation was found between CTG expansion and age of onset. Temporalis wasting and mental retardation were positively correlated with CTG expansion. No relationship was found between weakness distribution, electromyographic findings, other systemic features and CTG expansion. In this study of DM-1 in Iran, we found a correlation between CTG expansion and age of onset, temporalis wasting and mental disability. No correlation between CTG expansion and electrodiagnostic and other clinical findings were detected.

Occurrence of six virulence-associated genes in Arcobacter species isolated from various sources in Shiraz, Southern Iran.

In humans, arcobacters are associated with watery diarrhea and septicemia. Although, recently, more cases of diarrhea have been caused by Arcobacter species, very little is known about its pathogenesis. Therefore, the aim of this study was to evaluate the presence of six putative Arcobacter virulence genes (cadF, ciaB, cj1349, mviN, pldA, and tlyA), in a set of 113 Arcobacter butzleri, 40 Arcobacter cryaerophilus, and 15 Arcobacter skirrowii isolates that were recovered from various origins. The isolates were confirmed on the basis of polymerase chain reaction (PCR) amplification of genus and species specific PCR for determining three species. For confirmed isolates, PCR was carried out for the presence of virulence genes using specific primers. All A. butzleri isolates carried all six genes. For A. cryaerophilus and A. skirrowii, the cadF gene was detected just in 55 and 53.3%, ciaB in 97.5 and 86.6%, cj1349 in 45 and 60%, mviN in 90 and 80%, pldA in 32.5 and 13.3%, and tlyA in 37.5 and 40%, respectively. For A. cryaerophilus and A. skirrowii, the genes ciaB and mviN were significantly more prevalent than other virulence markers (P ≤ 0.05). The findings revealed that many of the important Arcobacter strains (86%) have these putative virulence genes which can be potential pathogenic properties for humans.

Distribution of nine virulence-associated genes in Campylobacter jejuni and C. coli isolated from broiler feces in Shiraz, Southern Iran.

To investigate the prevalence of nine virulence and toxin genes of Campylobacter, a total of 90 Campylobacter strains including 48 C. jejuni and 42 C. coli were recovered from chicken feces by cultivation methods. The isolates were identified on the basis of polymerase chain reaction (PCR) detection of 16SrRNA and multiplex PCR for determining two species. For confirmed strains, PCR was carried out for the presence of virulence genes using specific primers. Data were analyzed by SPSS software, version 12.0.1. The cadF gene and three genes associated with cytolethal distending toxin were present in 100% of isolates. Plasmid virB11 gene was not found in any of the Campylobacter isolates, and the prevalence of pldA, wlaN, iamA, and cgtB genes were 92.22%, 82.22%, 81.11%, and 22.22%, respectively. The findings revealed that the distribution of the majority of these genes were not dissimilar among Campylobacter species. The results emphasized that many of the pathogenic C. jejuni and C. coli may have these genes, and the Campylobacter strains with poultry origin have pathogenic potential properties for humans.

Association of Three Plasmid-Encoded spv Genes Among Different Salmonella Serotypes Isolated from Different Origins.

The virulence plasmid associated Salmonella plasmid virulence (spv) locus is strongly concomitant with strains that cause non typhoid bacteremia. The spv region contains three genes required for the virulence, the positive transcriptional regulator spvR and two structural genes spvB and spvC. The purpose of this study was to investigate the presence of these three genes among salmonella serotypes isolated from different sources. A collection of 60 salmonella serotypes from different sources were used. Polymerase chain reaction was carried out for the presence of these genes using specific primers. The prevalence of spvB, spvC, and spvR genes were 26 (43.3 %), 44 (73.3 %), and 28 (46.6 %), respectively. The findings revealed that the distribution of these genes was dissimilar among these serotypes. Many of the human pathogenic salmonella strains which can be transmitted by animals may have these genes and can be very injurious for public health.