Expression of virulence factor genes in co-infections with Trueperella pyogenes isolates and other bacterial pathogens; an in vivo study

Trueperella pyogenes is an opportunistic bacterial pathogen causing several infectious diseases, including metritis, mastitis and abscesses in domestic animals such as dairy cattle. Several virulence proteins are released by T. pyogenes strains contributing to the pathogenic and causing disease potential of this pathogen. So far, many aspects of T. pyogenes pathogenesis are unknown. In this study, expression levels of plo, fimA, nanH and cbpA genes encoding pyolysin, fimbriae, neuraminidase and collagen-binding protein, respectively in T. pyogenes isolated from totally 15 metritis, mastitis and cutaneous abscesses convenience samples in response to co-culture with other pathogens including E. coli, St. dysgalactiae, S. aureus, F. necrophorum and L. plantarum strains in mice study model have been investigated. We found that expression levels of plo, fimA, nanH and cbpA genes in T. pyogenes isolates in response to co-culture with F. necrophorum and E. coli were significantly increased; however, no significant changes was seen in the level of expression of these genes in the isolates in response to co-culture with St. dysgalactiae and S. aureus. Notably, expression of all virulence factor genes was suppressed in T. pyogenes in response to co-culture with L. plantarum. We observed that L. plantarum might be used to prevent infectious diseases caused by T. pyogenes

A quantitative prevalence of Escherichia coli O157 in different food samples using real-time qPCR method

Escherichia coli serogroup O157 is the main causative agent of several intestinal and extra-intestinal foodborne diseases in humans through consumption of low-dose contaminated foods such as milk, beef, and vegetables. To date, studies regarding the quantitative prevalence of E. coli O157 in foods are so limited. Therefore, this study aimed to evaluate the quantitative prevalence rate of E. coli serogroup O157 in raw milk (n = 144), vegetable salad (n = 174), and minced beef samples (n = 108) using the real-time qPCR SYBR green melting curve method targeting the rfbA gene. First, we evaluated the method and found a sensitive and specific qPCR assay with 1 log of CFU/ml detection limit to detect E. coli O157 (Tm = 80.3 ± 0.1°C). About 2.77%, 10.18%, and 9.19% of raw milk, minced beef, and vegetable salad samples, respectively, were contaminated with E. coli O157. Minced beef and vegetable salad samples were significantly more contaminated than raw milk samples. Population average of E. coli O157 in raw milk, minced beef, and vegetable salad samples were 2.22 ± 0.57, 3.30 ± 0.40, and 1.65 ± 0.44 log CFU/ml or gr, respectively. Significantly higher levels of population of E. coli O157 were observed in minced beef samples. Minced beef can be regarded as the main food in the transmission of this foodborne pathogen. Routine quantitative rapid monitoring is strongly suggested to be carried out to prevent foodborne diseases caused by E. coli O157

Oral abscess caused by chryseobacterium indologenes in ball python (Python regius) ::a case report

Chryseobacterium indologenes is an opportunistic pathogen isolated from human infections and, rarely, from some aquatic animals. A 3-year-old male ball python (Python regius) was admitted to the veterinary clinic by a pet owner because of acute respiratory and swallowing failure. During physical examinations, oral secretions and abscesses were observed in the mouth cavity and throat of the animal. After microbiological analysis including isolation, identification, and 16s rRNA sequencing, C. indologenes was detected as the main cause of the oral abscess in this case. Phylogenetic relatedness analysis showed a close relationship between this isolate and other strains isolated from human infections. Antimicrobial susceptibility testing revealed that the isolate was multi-drug resistant. However, it was very sensitive to minocycline, ceftazidime, and tetracycline. The patient was treated by antibiotic therapy and completely recovered after two weeks. To the best of our knowledge, this is the first incidence of C. indologenes in an oral abscess in a ball python. As a result we would consider this organism as an opportunistic animal pathogen with zoonotic potentiality

Cutaneous mycobacteriosis caused by Mycobacterium avium subspecies avium in a dog: Clinical findings, histopathological and molecular diagnosis and treatment

Abstract A 3‐year‐old castrated male Jack Russell Terrier with a history of cholestatic hepatitis was presented with a non‐painful and non‐pruritic, subcutaneous nodule following vaccine injection. The dog was otherwise healthy. The nodule was surgically removed. Upon gross inspection, a firm, glistening and loculated tissue was seen within the panniculus. Histopathologically, extensive pyogranulomatous panniculitis was diagnosed, composed of variably sized lipocyst surrounded by neutrophils and an outer zone of foamy epithelioid macrophages, neutrophils and occasional Langhans form giant cells. Although histopathologic findings were most compatible with mycobacteriosis, other infectious and sterile conditions were considered differential diagnoses. Gram, acid‐fast and periodic acid Schiff staining were negative. However, PCR analyses for mycobacterial rpoB gene and sequencing revealed infection with Mycobacterium avium subsp. avium. Empirical treatment was avoided through the 3‐week course of histopathological and molecular investigations. Considering that there were no newly formed cutaneous masses, marbofloxacin (2 mg/kg, PO q24h) was administered for 2 weeks, and clarithromycin was avoided due to its potential cholestatic hepatitis adverse effects. Eight months following surgery, there are no new or relapsing cutaneous lesions or disease‐specific complications. This suggests that marbofloxacin monotherapy may be sufficient in immunocompetent dogs with early detection of localized cutaneous mycobacteriosis lacking lymph node or organ involvement

Investigation of antimicrobial susceptibility and virulence factor genes in Trueperella pyogenes isolated from clinical mastitis cases of dairy cows

Trueperella pyogenes is an opportunistic pathogen causing important diseases including mastitis and metritis in domestic animals such as dairy cows leading to prominent economic losses in food production industry. The aim of this study was to investigate bacterial species, antimicrobial susceptibility, and presence of virulence factor genes and genotyping of T. pyogenes isolates associated with summer mastitis cases from 22 different farms around Tehran, Iran. Fifty-five percent of dairy cows with clinical mastitis symptoms was infected by T. pyogenesis indicated that this pathogen is the most important contributor to clinical mastitis in dairy cows in the present study. A significant correlation was illustrated between presence of virulence factor genes of isolated pathogen, biochemical patterns, and the utter infected types. Multidrug resistance susceptibility observed between isolates indicated the important need for prudent use of antimicrobials in treatment of mastitis caused by T. pyogenes and increased concerning of consumer health associated with recent problems of antimicrobial resistance. The categorization of isolates was implemented into seven different clonal related types by COX-PCR at 80% of similarity cutoff with significance relationship to clonal types, CAMP test result and sampling time and biochemical profile. Regarding to the results obtained at the present study, T. pyogenes can be considered as an important typically cause of purulent and acute form of clinical bovine mastitis and loss of dairy productivity. Further studies with more sample size and high-throughput omic methods in various sampling time and areas are suggested for study of this pathogen precisely. KEYWORDS antimicrobial susceptibility, dairy cow, Trueperella pyogenes, virulence factor gen

A quantitative prevalence of Escherichia coli O157 in different food samples using real‐time qPCR method

Escherichia coli serogroup O157 is the main causative agent of several intestinal and extra-intestinal foodborne diseases in humans through consumption of low-dose contaminated foods such as milk, beef, and vegetables. To date, studies regarding the quantitative prevalence of E. coli O157 in foods are so limited. Therefore, this study aimed to evaluate the quantitative prevalence rate of E. coli serogroup O157 in raw milk (n = 144), vegetable salad (n = 174), and minced beef samples (n = 108) using the real-time qPCR SYBR green melting curve method targeting the rfbA gene. First, we evaluated the method and found a sensitive and specific qPCR assay with 1 log of CFU/ml detection limit to detect E. coli O157 (Tm = 80.3 ± 0.1°C). About 2.77%, 10.18%, and 9.19% of raw milk, minced beef, and vegetable salad samples, respectively, were contaminated with E. coli O157. Minced beef and vegetable salad samples were significantly more contaminated than raw milk samples. Population average of E. coli O157 in raw milk, minced beef, and vegetable salad samples were 2.22 ± 0.57, 3.30 ± 0.40, and 1.65 ± 0.44 log CFU/ml or gr, respectively. Significantly higher levels of population of E. coli O157 were observed in minced beef samples. Minced beef can be regarded as the main food in the transmission of this foodborne pathogen. Routine quantitative rapid monitoring is strongly suggested to be carried out to prevent foodborne diseases caused by E. coli O157

Repetitive sequences based on genotyping of Candida albicans isolates obtained from Iranian patients with human immunodeficiency virus

Objective(s): Candidiasis infection caused by Candida albicans has been known as a major problem in patients with immune disorders. The objective of this study was to genotype the C. albicans isolates obtained from oral cavity of patients with positive human immunodeficiency virus (HIV+) with or/and without oropharyngeal candidiasis (OPC). Materials and Methods:A total of 100 C. albicans isolates from Iranian HIV+patients were genotyped using specific PCR primers of the 25S rDNA and RPS genes. Results: The frequencies of genotypes A, B and C which were achieved using 25S rDNA , were 66, 24 and 10 percent, respectively. In addition, genotypes D and E were not found in this study. Each C. albicans genotype was further classified into four subtypes (types 2, 3, 2/3 and 3/4) by PCR amplification targeting RPS sequence. Conclusion:In general, genotype A3 constituted the majority of understudy clinical isolates obtained from oral cavity of Iranian HIV+ patients

Morganella morganii infection in hirudo medicinalis (Iran) ::a case report

Medicinal leeches (Hirudo medicinalis) are used in surgical and non-surgical manners. Morganella morganii is an opportunistic and zoonotic pathogenic bacterium causing serious clinical complications. In this study, we isolated, discovered and characterized M. morganii-infected H. medicinalis. We detected and identified M. morganii in all inflamed and swollen Hirudo medicinalis samples. The 16S rRNA sequence of the isolates confirmed all strains of M. morganii. All strains were sensitive to Ceftriaxone, Ceftiofur, Danofloxacin, Ciprofloxacin, Enrofloxacin, Oxytetracycline, and Meropenem and were resistant to Erythromycin, Amoxicillin, Ampicillin, Cefazolin, Colistin, Penicillin G, and Lincomycin. This pathogenic bacterium is a zoonotic pathogen, and monitoring the prevalence rate of this bacteria is strongly necessary for leeches used in human medical treatment and care. Finally, all infected leeches were treated successfully in this case report study

Isolated Bacteria from the Uteri of Camels with Different Reproductive Backgrounds: A Study on Sampling Methodology, Prevalence, and Clinical Significance

The objectives of this study were to comparatively identify the common bacterial isolates from the uteri of camels coming from different reproductive backgrounds after standardizing the sampling method and to investigate the association of clinically measurable parameters with uterine colonization by these isolates. The uterine samples from 856 dromedary camels yielded a total of 17 different bacterial species with a higher proportion of sub-fertile camel uteri being colonized by bacteria (66.6%) as compared to nulliparous, recently calved, and those with unknown reproductive history combined (44.2%; p p > 0.05). While certain strains were more likely to be obtained from the uteri of the sub-fertile group (p p > 0.05). In conclusion, a relatively high bacterial load can be identified from the uteri of both sub-fertile and normal dromedary camels, with a higher frequency among the former. The uterine ultrasonography and evaluation of the body condition score can help in identifying the camels in which uterus is contaminated by bacteria

Complete genome sequence of Trueperella pyogenes strain Arash114, isolated from the uterus of a water buffalo (Bubalus bubalis) in Iran

Objective: Trueperella pyogenes has been considered a major causative agent of metritis, abortion, and death in a broad range of domestic and wild animals, including cattle, swine, sheep, goats, camels, buffalo, deer, antelopes, reptiles, and birds. Data description: Here, we report the complete chromosome sequence of Trueperella pyogenes strain Arash114, isolated from the uterus of a water buffalo (Bubalus bubalis) died due to the infection caused by this pathogen. The genome assembly comprised 2,338,282 bp, with a 59.5% GC content. Annotation of the genome showed 46 tRNA genes, 6 rRNA, 1 CRISPR and 2059 coding sequences. Also, several genes coding for antimicrobial resistance such as tetW and virulence factor including plo, nanH, nanP, cbp and 4 fimbrial proteins were found. This study will advance our knowledge regarding the metabolism, virulence factors, antibiotic resistance and evolution of Arash114 strain and serve as an appropriate template for future researches. Keywords: Complete genome sequencing; Trueperella pyogenes; Uterus infection; Water buffalo