Molecular detection and identification of hemotropic Mycoplasma species in dogs and their ectoparasites in Iran.

Hemotropic Mycoplasma species are vector-borne bacteria that attach and grow on the surface of erythrocytes in various mammals, yet reports of canine hemoplasmosis in Iran are scarce. The aim of this study was molecular detection and identification of hemoplasmas in the blood of dogs (n = 370) from five provinces of Iran and ectoparasites infesting them including Ctenocephalides canis and Pulex irritans fleas, Rhipicephalus sanguineus sensu lato ticks, Heterodoxus spiniger lice and Hippobosca longipennis keds. Hemotropic Mycoplasma spp. pathogens were detected using genus-specific conventional PCRs, and subsequently identified using species-specific PCRs for Mycoplasma haemocanis (Mhc), and Candidatus Mycoplasma haematoparvum (CMhp). Sanger sequencing was then performed to confirm the species. Correlation of infection and risk factors (geographical area, keeping condition, body condition, sex, age, ectoparasite infestation) were analyzed. In total, 210 dogs (56.7%) were tested PCR-positive for hemotropic Mycoplasma spp. Species-specific PCR and sequencing revealed infection with Mhc in 17.8%, with CMhp in 7.02% and co-infection in 31.9% of dogs. Flea infestation, poor body condition, and being older than 3-years-old correlated with hemoplasmosis. In ectoparasites, DNA of hemoplasmas were detected only in fleas i.e. Mhc in P. irritans, CMhp in P. irritans and C. canis, and co-infection in C. canis. To our knowledge, this is the first large-scale molecular epidemiology study of canine hemoplasmosis in Iran. Considering the high prevalence of canine hemoplasmosis all over the country including potentially zoonotic CMhp, effective ectoparasite control strategies, regular examination of dogs, successful chemoprophylaxis and public awareness strategies are advocated.

Molecular detection and identification of Bartonella species in cats from Hamedan and Kermanshah, Western Iran.

Bartonella species are emerging vector-borne zoonotic pathogens which infect a wide range of domestic and wild animals as well as humans. Cats are the primary reservoir hosts for several zoonotic Bartonella spp., the most common being B. henselae the causative agent of cat scratch disease. Despite the important role of cats in the epidemiology of bartonellosis, there is limited information about the prevalence and species infecting cats in Iran. The aim of present study was molecular detection and identification of Bartonella species in cats from two western provinces Hamedan and Kermanshah. From December 2018 to February 2021, 87 cats (n = 26 from Hamedan, n = 61 from Kermanshah) were examined clinically, their bodies were searched for collection of ectoparasites, and cephalic or saphenous blood specimens were collected. Genomic DNA was extracted from blood specimens and conventional PCRs targeting the rpoB, and ITS regions of Bartonella spp. were performed. Positive samples were sequenced and analysed phylogenetically. Bartonella DNA was detected in 11/87 cats (12.64 %). Sequencing results revealed the presence of B. henselae in cats from Hamedan, and B. clarridgeiae and B. henselae in cats from Kermanshah. A statistical association between cat origin and the prevalence of Bartonella spp. was observed in the studied population. This study confirms for the first time the circulation of Bartonella spp. in cats in two western Iranian provinces. Prevention strategies e.g. ectoparasites control, and regular examination of pet and urban cats are suggested for minimising Bartonella infection in cats and subsequently in humans.

Antibiotic resistance and virulence factor gene profile of A. hydrophila isolated from carp (Cyprinidae) suspected with hemorrhagic septicemia in Gilan, Iran.

The present study was conducted to determine the antibacterial resistance profile of Aeromonas hydrophila (n = 42) isolated from the 100 hemorrhagic septicemia-suspected carp in Gilan, Iran. The prevalence of class 1 and 2 integrons, antibiotic resistance genes (ARG) and virulence factor genes (VFG) among these isolates was investigated using PCR. Also, the possible association between the presence of VFGs and the antibiotic resistance profile of isolates was assessed. The majority of A. hydrophila isolates (83·33%) exhibited multi-drug resistance (MDR) profile, and all isolates were resistant to clindamycin, while all isolates were susceptible to amikacin. intI1 and intI2 gene was found in 26·2 and 4·8% isolates, respectively. This is the first report of the presence of the intI2 gene in A. hydrophila isolates in Iran. The blaTEM (40·5%) and tetA (33·3%) genes were found as the predominant ARGs. The most frequently detected VFGs were lip and ahh1(90·5%), while the examined isolates carrying at least three VFGs and the most prevalent VFGs profile was ast+, act+, alt+, ahhl+, aerA+, ahyB+ and lip+. The results of this study indicate a positive association between the presence of VFGs and antibiotic resistance, and most MDR A. hydrophila isolates showed high frequencies of VFGs.

High Prevalence of Multidrug Resistance and Biofilm-Formation Ability Among Avian Escherichia coli Isolated from Broilers in Iran.

The present study was conducted to determine the antimicrobial resistance pattern and biofilm-formation ability in 100 Avian-Pathogenic Escherichia coli (APEC) isolated from colibacillosis-suspected broilers and 100 Avian Fecal E. coli (AFEC) isolates from healthy broilers in Hamedan, Iran. All isolates were screened by polymerase chain reaction for antimicrobial resistance genes, class 1 and 2 integrons, and biofilm-associated genes. Besides, we assessed the possible relationship between biofilm-formation ability antibiotic resistance patterns, genetic background, and the pathogenicity of APEC strains. 81% of APEC and 73% of AFEC isolates showed multidrug resistance (MDR) phenotype; in addition, 45% of the APEC and 21% of the AFEC strains showed biofilm-formation ability. This is the first report of the biofilm formation ability in E. coli isolated from broilers in Iran. The most prevalent antibiotic resistance gene in APEC strains was tetA (68%), followed by sul1 (63%), dfrA1-like (51%), and blaTEM (30%), whereas in AFEC strains, the frequencies of the antibiotic resistance genes were tetA (63%), sul1 (58%), dfrA1-like (49%), and blaTEM (22%). Out of 81 MDR APEC isolates, 53 (65.4%) and 38 (46.91%) isolates were positive for intI1 and intI2 genes, respectively. In the AFEC strains intI1 and intI2 genes were presented in 57 and 33 isolates, respectively. All APEC isolates belonging to phylogenetic groups B1, B2, and C were MDR. The results of the present study indicate that isolates with biofilm-forming ability show more MDR properties and probably have more pathogenicity to broilers.

Molecular Detection of Zoonotic Pathogens in the Blood and Tissues of Camels (Camelus dromedarius) in Central Desert of Iran.

Dromedary camels (Camelus dromedarius) play a major economic role in many countries in Africa and Asia. Although they are resistant to harsh environmental conditions, they are susceptible to a wide range of zoonotic agents. This study aimed to provide an overview on the prevalence of selected zoonotic pathogens in blood and tissues of camels in central Iran. Blood, liver, portal lymph node, and brain were collected from 100 apparently healthy camels at a slaughterhouse in Qom city to assess the presence of DNA of Brucella spp., Trypanosoma spp., Coxiellaburnetii, and Bartonella spp. PCR products were sequenced bidirectionally and phylogenetic analyses were performed. Eleven percent of camels tested positive for Brucellaabortus (3%) and Trypanosomaevansi (8%). Coxiellaburnetii and Bartonella spp. DNA was not detected. Our data demonstrate that camels from Iran contribute to the epidemiology of some zoonotic pathogens. Performing proper control strategies, such as vaccination of camels and humans in contact with them, test-and-slaughter policy, and education of the general population is necessary for minimizing the risk of zoonotic infection.

Serogrouping, phylotyping, and virulence genotyping of commensal and avian pathogenic Escherichia coli isolated from broilers in Hamedan, Iran.

In the present study, 100 Avian-Pathogenic Escherichia coli (APEC) isolates from colibacillosis-suspected broilers and 100 Avian Faecal Escherichia coli (AFEC) isolates from healthy broilers in Iran were examined by PCR for confirmation of their serogroups and phylogenetic background, and their association with ten virulence-associated genes (VAG) including fimC, iutA, chuA, sitA, iss, cvaA/B, hylA, stx1, stx2, and yjaA. Serogroups O78, O1, O2 and O18 were the prominent strains including 54 % of the APEC and 23 % of the AFEC strains. At phylotyping, the majority of APEC strains belonged to phylogenetic group E (22 %) while for the AFEC strains, half of the isolates were not assigned to any group but the predominant phylogroup was E (27 %). Virulence genotyping, revealed that the predominant VAGs were iutA (97 %), fimC (87 %) and iss (84 %) among APEC strains, and fimC (95 %), iss (93 %) and sitA (87 %) in AFEC strains. This is the first time that phylogroup E is described as predominant phylogroup among APEC strains also, this is the first report on the presence of the stx1 gene in APEC strains isolated from broilers in Iran. The results of the present study indicate that VAGs are more prevalent in APEC strains belonging to O2 and O78 serogroups, also phylogroups E and D have more frequency of VAGs than other phylogroups. Therefore, the APEC strains belonging to O2 and O78 serogroups and phylogroups E and D probably have more pathogenicity to broilers.

High Prevalence of Bartonella sp. in Dogs from Hamadan, Iran.

Bartonellae are emerging vector-borne pathogens infecting various domestic and wild mammals. Blood samples were collected from 66 dogs at two locations near Hamedan, Iran. Twenty dogs were rescued stray dogs and 46 dogs were from a breeding colony, with many of them infested with fleas, ticks, or lice. Serology was performed using an indirect immunofluorescent antibody test for Bartonella henselae, Bartonella clarridgeiae, and Bartonella vinsonii subsp. berkhoffii. Seroprevalence was 74.2% (range: 65.2-95%). Bartonella DNA amplification and sequencing identified B. vinsonii subsp. berkhoffii type III in seven dogs, including five rescued dogs. Two dogs were infected with Bartonella rochalimae and three dogs with Candidatus B. merieuxii, including two of the stray dogs coinfected with Bartonella vinsonii subsp. berkhoffii. Rescued stray dogs were 10 times (odds ratio (OR) = 10.13, 95% CI: 1.24-82.7; P = 0.03) more likely to be seropositive and eight times (OR = 8.82, 95% CI: 2.68-29.11; P = 0.0004) more likely to be flea-infested than breeding dogs, confirming that arthropod infestation is a major risk factor for these infections.

Biological function of a polysaccharide degrading enzyme in the periplasm.

Carbohydrate polymers are industrially and medically important. For instance, a polysaccharide, alginate (from seaweed), is widely used in food, textile and pharmaceutical industries. Certain bacteria also produce alginate through membrane spanning multi-protein complexes. Using Pseudomonas aeruginosa as a model organism, we investigated the biological function of an alginate degrading enzyme, AlgL, in alginate production and biofilm formation. We showed that AlgL negatively impacts alginate production through its enzymatic activity. We also demonstrated that deletion of AlgL does not interfere with polymer length control, epimerization degree or stability of the biosynthesis complex, arguing that AlgL is a free periplasmic protein dispensable for alginate production. This was further supported by our protein-stability and interaction experiments. Interestingly, over-production of AlgL interfered with polymer length control, suggesting that AlgL could be loosely associated with the biosynthesis complex. In addition, chromosomal expression of algL enhanced alginate O-acetylation; both attachment and dispersal stages of the bacterial biofilm lifecycle were sensitive to the level of O-acetylation. Since this modification also protects the pathogen against host defences and enhances other virulence factors, chromosomal expression of algL could be important for the pathogenicity of this organism. Overall, this work improves our understanding of bacterial alginate production and provides new knowledge for alginate production and disease control.