Molecular detection and phylogenetic analysis of Borrelia Spp. In blood samples of cats and dogs by the nested-PCR method in West Azerbaijan Province, Iran.

The objective of this study was to investigate the presence and genetic attributes of Borrelia spp. in cats and dogs from the West Azerbaijan Province, located in the northwest of Iran. A total of 250 blood samples from cats and 300 blood samples from dogs were collected, and information regarding their age, sex, breed, ownership status, sampling time and region was recorded. The identification of positive samples was accomplished through nested-PCR and sequencing, with subsequent analysis of the gene sequences conducted using BioEdit software. The gene sequences for Borrelia spp. in this study showed 100% similarity to reference sequences in the GenBank® database. Phylogenetic trees were built using MEGA11. The outcomes indicated that among 250 blood samples from cats, 48 (19.2%) tested positive for Borrelia spp. gene, with a CI from 14.8 to 24.53% for cats. Similarly, out of 300 blood samples from dogs, 45 (15%) tested positive for the Borrelia spp. gene, with a CI from 11.4 to 19.48% for dogs.

Molecular detection and phylogenetic analysis of Borrelia spp. from sheep and goats blood samples in West Azerbaijan province, Iran.

Borrelia species are spirochetes transmitted by ticks that are important in human and animals. In most countries, there is still no molecular epidemiology of borreliosis in ruminants. This study was aimed to evaluate the existence of Borrelia spp. DNA in the blood samples of small ruminants using polymerase chain reaction (PCR) method in West Azerbaijan Province, Iran. To detect Borrelia spp. DNA, about 1,018 ruminants (456 goats and 562 sheep) blood samples were examined from different bioclimatic regions in West Azerbaijan province, Iran. The DNA extracting and PCR were conducted. In sheep, the following prevalence rates were respectively obtained for the 16S rRNA, 5S - 23S rRNA and ospA genes: 3.55% (20/562), 2.13% (12/562) and 0.88% (5/562). And so, the prevalence rates of the genes in goats were 0.87% (4/456) for 5S - 23S rRNA gene, 1.75% (8/456) for 16S rRNA gene and 0.65% (3/456) for ospA gene. The prevalence of Borrelia spp. was significantly different in small ruminants based on the farms and localities. The sheep and goats in humid areas (north of West Azerbaijan) were infected statistically more than those in sub-humid areas (south of West Azerbaijan). It is demonstrated that host species like sheep and goats may have a key role in natural Lyme disease cycles and other borreliosis diseases in Iran.

Genomic detection and phylogenetic analysis of Bartonella quintana in pet cats from Urmia City, Northwest Iran.

The aim of this study was to investigate the presence and genetic characteristics of Bartonella quintana in pet cats from Urmia City, located in the northwest of Iran. Blood samples were collected from 200 cats, and their age, gender, and breed were noted. Nested-PCR and sequencing were used to identify B. quintana in positive samples, and the ftsZ gene sequences were analyzed using BioEdit software. The gene sequence obtained in this study exhibited 100.00 % similarity to reference sequences in the GenBank® database, and a phylogenetic tree was constructed using MEGA11. The results revealed that 15 % of the cats (30 out of 200 blood samples) tested positive for the B. quintana gene, with a 95 % confidence interval of 10.71 % to 20.61 %.

Molecular Detection of Borrelia spp. in Ticks of Sheep and Goats by Nested PCR Method in West Azerbaijan Province, Iran.

Background: Ticks are an important vector among arthropods associated with serious medical and veterinary problems. In this research, we investigate Borrelia species in ticks isolated from the surface of livestock (sheep and goat) in different regions of West Azerbaijan province. Materials and Methods: Polymerase chain reaction (PCR) was performed using specific primers targeting Borrelia spp. genes. Borrelia spp. was identified through PCR. The positive PCR products were sent to Pishgam Company for sequencing. Sequenced data were analyzed, and phylogenetic analysis was performed using maximum likelihood method in MEGA V.10. Results: The detection rate of Borrelia spp. for 16srRNA gene 69 (n = 542; 12.7%; 95%Cl: 10.1%-15.8%), 42 (n = 542; 7.7%; 95%Cl: 5.78%-10.1%) positive on 5S-23SrRNA gene and ospA gene 4 (n = 542; 0.74%; 95%Cl: 0.29%-1.88%). Conclusion: These results are the first report in Iran to identify Borrelia spp. These results of the study showed that the Borrelia spp. in hard ticks detected by PCR and it was negative to soft ticks. it is important in public health implications at the studied areas.

Genomic detection of Coxiellaburnetii based on plasmid genes in horses.

Q fever is a worldwide zoonosis caused by an obligate intra-cellular pathogen called Coxiella burnetii affecting a broad range of animal hosts including horses. Most of the isolates found carry plasmids which genetic studies of C. burnetii strains suggest a critical role in C. burnetii survival. The correlation between an isolated plasmid type and the chronic or acute nature of the disease has always been controversial. This study was conducted to investigate the prevalence of C. burnetii QpH1 and QpDG plasmids in horses and assess the potential role of these species as reservoirs of infection and transmission. Nested-polymerase chain reaction (PCR) assays were performed on 320 blood serum samples drawn from horses in West Azerbaijan province, Iran, in 2020. In total, 26 (8.13%) Q fever-positive samples based on containing the IS1111 gene were tested by nested-PCR approach to amplify QpH1 and QpDG plasmid segments. The QpH1 and QpRS plasmid-specific sequences were identified in 19 (73.07%) and none in the serum samples, respectively. According to the present study, the age of the animal can be considered as an important risk factor for the prevalence of C. burnetii; but, the season, sex, and breed of the horse had no effect on the prevalence of disease. The results indicate that nested-PCR method could be suitable for routine diagnosis, to gather new information about the shedding of C. burnetii, and to improve the knowledge of contamination routes.

Alum and metoclopramide synergistically enhance cellular and humoral immunity after immunization with heat-killed Salmonella typhimurium vaccine.

Typically, the killed form of microorganisms in combination with alum does not produce strong cellular immune responses. A recent investigation has indicated the role of dopamine D2 receptor antagonists like metoclopramide in reducing the polarization of immune responses toward Th2 immunity. This study was performed to evaluate the effects of a combination of alum and metoclopramide on the induction of cellular and humoral immunity in response to a heat-killed preparation ofSalmonella typhimurium(HKST). Wistar rats were immunized with the HKST vaccine alone or in combination with alum, metoclopramide, or the alum-metoclopramide mixture twice with a two-week interval. Fourteen days after the last vaccination, immune responses against S. typhimurium and the protective potential of the vaccines were assessed. The combination of alum and metoclopramide as an adjuvant augmented the potential of the HKST vaccine to enhance lymphocyte proliferation, delayed-type hypersensitivity reaction, and antibody titer. These results were concurrent with the polarization of immune response towards the Th1 response and improving protective immunity against S. typhimurium. Overall, the combination of alum and metoclopramide as an adjuvant synergistically enhanced cellular and humoral immunity after immunization with the HKST vaccine.

Phylogenetic analysis of Escherichia coli isolated from broilers with colibacillosis based on gyrA gene sequences.

Escherichia coli isolates from chickens with colibacillosis were assigned to phylogenetic groups based on multiplex polymerase chain reaction (PCR) and antibacterial resistance of E. coli belonging to these groups was examined. Furthermore, the gyrA gene of isolates was sequenced and a phylogenetic tree was generated. A total of 84 E. coli isolates were grouped using multiplex PCR of TSPE4.C2, chuA, yjaA, and gadA molecular markers. Four phylogenetic groups were identified with strains divided as follows: 16 in group A (19.05%), 17 in group B1 (20.24%), 23 in group B2 (27.38%), and 28 in group D (33.33%). Escherichia coli isolates belonging to phylogenetic groups B2 and D were resistant to Soltrim and Flumequine unlike the majority of E. coli isolates that belonged to groups A and B1, and which were susceptible to these antibiotics. The phylogenetic results based on gyrA gene sequences from multiplex PCR revealed that E. coli phylogenetic grouping was in accordance with the clusters obtained in the phylogenetic tree. In conclusion, the comparative sequence analysis of gyrA sequences provides a firm framework for an accurate classification of E. coli and related taxa and may constitute a pertinent phylogenetic marker for E. coli.

Les isolats d'Escherichia coli provenant de poulets avec colibacillose ont été assignés à des groupes phylogénétiques sur la base d'une réaction d'amplification en chaine par la polymérase multiplex (ACP) et la résistance antimicrobienne des E. coli appartenant à ces groupes a été examinée. De plus, le gène gyrA des isolats a été séquencé et un arbre phylogénétique a été généré. Un total de 84 isolats a été groupé à l'aide de l'ACP multiplex utilisant les marqueurs moléculaires TSPE4.C2, chuA, yjaA et gadA. Quatre groupes phylogénétiques ont été identifiés et les souches réparties comme suit: 16 dans le groupe A (19,05 %), 17 dans le groupe B1 (20,24 %), 23 dans le groupe B2 (27,38 %), et 28 dans le groupe D (33,33 %). Les isolats d'E. coli appartenant aux groups phylogénétiques B2 et D étaient résistants au Soltrim et à la fluméquine contrairement à la majorité des isolats d'E. coli appartenant aux groupes A et B1, qui étaient sensibles à ces antibiotiques. Les résultats phylogénétiques basés sur les séquences du gène gyrA provenant de l'ACP multiplex ont révélé que le regroupement phylogénétique des E. coli était en accord avec les groupes obtenus dans l'arbre phylogénétique. En conclusion, l'analyse comparative des séquences gyrA fourni un patron solide pour une classification précise des E. coli et taxons associés et pourrait constituer un marqueur phylogénétique pertinent pour les isolats d'E. coli.(Traduit par Docteur Serge Messier).

Prevalence and molecular characterization of staphylococci isolated from sheep with subclinical mastitis in West-Azerbaijan province, Iran.

This study was conducted to investigate the prevalence of subclinical mastitis caused by Staphylococcus spp. in ewes in West-Azerbaijan province of Iran. Molecular characterization of isolated Staphylococcus spp. from diseased ewes were performed using polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) and DNA sequencing of glyceraldehyde-3-phosphate dehydrogenase (gap) gene. Also, antibiotic resistance of staphylococcal isolates against different antibiotics was investigated. A total number of 900 milk samples from 450 native ewes in their mid-lactation period were examined by the California mastitis test (CMT). The CMT positive samples were cultured and bacteria were isolated from 86 (9.50%) glands and 74 (16.40%) ewes. The prevalence of subclinical mastitis in the examined ewes was 16.40%. Microbiological analysis of milk samples revealed that 27 out of 74 sheep with subclinical mastitis were infected with Staphylococcus spp. Amplification of gap gene of 27 Staphylococcus isolates generated a single amplicon of 933 bp in size confirming that isolates were belonged to Staphylococcus genus. Digestion of PCR products by AluI endonuclease generated different RFLP patterns for each species. Nucleotide sequencing of gap gene followed by phylogenetic analysis showed that the most dominant Staphylococcus species were S. epidermidis, S. xylosus and S. chromogenes. Staphylococcal isolates showed the highest resistance to penicillin and ampicillin. In conclusion, Staphylococcus species, except for the southern parts of the province, play an important role in the development of subclinical mastitis in sheep in West-Azerbaijan province of Iran. Also, chloramphenicol, ciprofloxacin and neomycin are the most effective antibiotics for treatment of this disease.

In vitro antibacterial activities of ethanol extract of iranian propolis (EEIP) against fish pathogenic bacteria (Aeromonas hydrophila, Yersinia ruckeri & Streptococcus iniae)

The ''in vitro'' antibacterial activity of ethanol extract of propolis (EEIP) from Urmia, Iran was investigated against three prevalent species of fish bacterial pathogens including: Aeromonas hydrophila LMG 3770, Yersinia ruckeri LMG 3279 and Streptococcus iniae LMG 14520. In this study two standard susceptibility testing techniques (Micro-broth dilution method and Agar-well diffusion method) were used to evaluation of the antibacterial activity of EEIP against the mentioned micro-organisms. Also the chemical composition of propolis was determined by the method of Gas chromatography-mass spectrometry (GC-MS). Twenty-six compounds were identified by gas chromatography-mass spectrometry analysis. Results showed Chemical composition of EEIP contained significant amounts of flavonoids, Sesquiterpenes - mainly Eudesmol and Caryophyllene oxide - aromatic acid, and low amounts of aldehydes and triterpens. Furthermore the ethanol extract of propolis inhibited the growth of all examined micro-organisms with the highest antimicrobial activity against Gram-positive bacteria Streptococcus iniae. Ethanol did not influence the antimicrobial effect of EEIP. These antibacterial properties would warrant further studies on the clinical applications of propolis in aquaculture field.

In vitro antibacterial activities of ethanol extract of iranian propolis (EEIP) against fish pathogenic bacteria (Aeromonas hydrophila, Yersinia ruckeri & Streptococcus iniae).

The "in vitro" antibacterial activity of ethanol extract of propolis (EEIP) from Urmia, Iran was investigated against three prevalent species of fish bacterial pathogens including: Aeromonas hydrophila LMG 3770, Yersinia ruckeri LMG 3279 and Streptococcus iniae LMG 14520. In this study two standard susceptibility testing techniques (Micro-broth dilution method and Agar-well diffusion method) were used to evaluation of the antibacterial activity of EEIP against the mentioned micro-organisms. Also the chemical composition of propolis was determined by the method of Gas chromatography-mass spectrometry (GC-MS). Twenty-six compounds were identified by gas chromatography-mass spectrometry analysis. Results showed Chemical composition of EEIP contained significant amounts of flavonoids, Sesquiterpenes - mainly Eudesmol and Caryophyllene oxide - aromatic acid, and low amounts of aldehydes and triterpens. Furthermore the ethanol extract of propolis inhibited the growth of all examined micro-organisms with the highest antimicrobial activity against Gram-positive bacteria Streptococcus iniae. Ethanol did not influence the antimicrobial effect of EEIP. These antibacterial properties would warrant further studies on the clinical applications of propolis in aquaculture field.

Antibacterial and antifungal activity of Iranian propolis against Staphylococcus aureus and Candida albicans.

Propolis samples from West North region of Iran were studied for their antibacterial (against Staphylococcus aureus) and antifungal (against Candida albicans) activities. In this article, yield of extracts and their pH values were measured. Antibacterial and antifungal activities of Ethanol-Extracted Propolis (EEP) were investigated by Petri dish bioassay method. Dilutions of EPP in agar with serial concentrations ranging from 0/04 to 10% (W/V) were prepared and antimicrobial activities were determined as Minimal Inhibitory Concentrations (MIC). All samples were active against the fungal and bacterial test strains. MIC values for different propolis samples against Staphylococcus aureus were, respectively 4, 3 and 1.5% (W/V) and against Candida albicans were, respectively 2, 4 and 3% (W/V).