Phylogenetic Analysis of Brucella melitensis Strains Isolated from Humans Using 16S rRNA Sequencing and Multiple Locus Variable Number of Tandem Repeats Analysis-16.

Background: Brucellosis is the most important public health problem worldwide, and the annual incidence of the disease in humans is 2.1 million. The Brucella genome is highly conserved, with over 90% similarity among species. The aim of this study was to perform species-level identification of Brucella spp. strains isolated from humans diagnosed with brucellosis and to further investigate the phylogenetic relationships using multiple locus variable number of tandem repeats analysis (MLVA)-16 and 16S rRNA sequencing analysis. Materials and Methods: Brucella spp. was isolated from the blood cultures of 54 patients who tested positive for brucellosis through serological examinations. Real-time PCR was used to identify the isolates in species, and the genus level of Brucella was confirmed with 16S rRNA. All isolates were subjected to phylogenetic analysis using variable number of tandem repeat analysis with multiple loci. Results: Subsequent analysis via real-time PCR confirmed these isolates to be of the Brucella melitensis species. The 16S rRNA sequence analysis showed 100% homogeneity among the isolates. MLVA revealed the formation of five different genotypic groups. While two groups were formed based on the 16S rRNA sequence analysis, five groups were formed in the MLVA. Conclusions: The study concluded that 16S rRNA sequence analysis alone did not provide sufficient discrimination for phylogenetic analysis but served as a supportive method for identification. MLVA exhibited higher phylogenetic power. The widespread isolation of B. melitensis from human brucellosis cases highlights the importance of controlling brucellosis in small ruminants to prevent human infections.

Molecular prevalence of Coxiella burnetii in cheese samples: Systematic review and meta-analysis.

BACKGROUND: Cheese is a popular dairy product consumed worldwide, and it has been implicated as a source of Coxiella burnetii infections. OBJECTIVES: The present study aimed to describe the molecular prevalence and source analysis of C. burnetii in cheese samples. METHODS: A systematic literature search was conducted using the Medline/PubMed, Science Direct, Web of Science, Scopus, and Google Scholar databases to identify studies reporting the molecular prevalence of C. burnetii in cheese samples. The pooled prevalence of C. burnetii in cheese samples was estimated using a random-effects model. RESULTS: A meta-analysis was conducted using the mean and standard deviation values obtained from 13 original studies. The overall molecular prevalence of C. burnetii in cheese was estimated to be 25.2% (95% confidence interval [CI]: 13.1%-39.7%). The I2 value of 96.3% (CI95% 94.9-97.3) suggested high heterogeneity, with a τ2 of 0.642 (CI95% -0.141 to 0.881), and an χ2 statistic of 323.77 (p < 0.0001). CONCLUSIONS: In conclusion, our meta-analysis provides a thorough assessment of the molecular prevalence and source analysis of C. burnetii in cheese samples.

Detection of Coxiella burnetii and characterisation by multiple-locus variable-number tandem repeat analysis in bovine bulk tank milk samples.

Coxiella burnetii is the aetiological agent of Q fever, which is highly prevalent in Turkiye, but information on the genetic profiles of the bacterium is limited. This study aimed to investigate the presence of C. burnetii in bovine bulk tank milk (BTM) samples by polymerase chain reaction (PCR) and to investigate the genotypes by means of multiple-locus variable-number tandem repeat analysis (MLVA). A total of 25 markets that sold raw cow's milk were analysed by conventional PCR analysis. An MLVA analysis was performed at six loci, namely MS23, MS24, MS27, MS28, MS33, and MS34, to determine the genotypic variations of C. burnetii found in the positive DNA samples. The DNA of C. burnetii was detected in 16% of the BTM samples. The C. burnetii strains identified in the bovine milk samples collected in this study were found to belong to the same genotypic group as those detected in the bovine milk samples gathered in Greece. As a result, both the presence and genotyping studies of C. burnetii on the BTM samples in Turkiye will contribute to the determination of the geographical distribution of the agent.

Investigation of virulence factors, phylogenetic grouping, multiple-locus variable number tandem repeat analysis, and antimicrobial susceptibility of E. coli isolated from aborted bovine fetal tissue.

Escherichia coli is an important microorganism for cattle breeding. The aim of this study was to investigate the presence of phylogenetic groups, virulence factors, genotyping with multi-locus variable tandem repeat analysis (MLVA), and susceptibility to commonly used antimicrobial agents in E. coli strains isolated from aborted bovine fetal samples. In this study, phylogrouping and various virulence genes were analyzed by PCR in E. coli strains isolated from 637 bovine fetal tissue samples. Consequently, E. coli was isolated and identified in 24 samples in culture. Of the 24 isolates identified as positive, 12.5% were defined as group A, 83.3% as B1, and 4.2% as group B2. Of the E. coli isolates, virulence factor fimH was identified in eight (33.3%), traT in 15 (62.5%), ompT in five (20.8%), CNF1 in one (4.16%), and CNF2 in six (25%). Seven genotypic groups were determined as a result of the analysis with the MLVA 10 method. According to the antimicrobial susceptibility test results, high resistance was determined against amoxicillin/clavulanic acid and oxytetracycline. In conclusion, strains of E. coli containing CNF1, CNF2, fimH, traT, and ompT virulence factors can be associated with bovine abortions. It is noteworthy that the dominant phylogenetic group B1 has been observed in cases of cattle abortions.

Genotyping of Brucella isolates from animals and humans by Multiple-Locus Variable-number Tandem Repeat Analysis (MLVA).

This study investigates country-wide genotype variations through the genotyping of Brucella strains isolated from domestic ruminants and humans. The Brucella spp. isolated from samples taken from animals and humans were first identified as B. abortus and B. melitensis by real-time PCR, and the MLVA-16 approach was then used for the genotyping of the identified isolates. For the study, 416 Brucella spp. were isolated from aborted fetus samples examined between 2018 and 2021, and 74 Brucella spp. from infected humans. Of the 74 human isolates analyzed, 1.3% were identified as B. abortus and 98.7% (73/74) as B. melitensis. The MLVA-16 typing method revealed 30 clonal groups for B. abortus and 37 clonal groups for B. melitensis from which the dominant genotypes and similarities with human isolates in Türkiye were determined.

Molecular epidemiology of Coxiella burnetii detected in humans and domestic ruminants in Turkey.

Q fever is a zoonotic disease that is known to be widespread throughout the world by many researches since its discovery in 1935 and it is important in terms of animal and public health. Coxiella burnetii, which is the etiological agent of the disease, is an obligate intracellular pathogen. While the disease generally manifests itself with abortion in animals, disease manifests as atypical pneumonia or granulomatous hepatitis in the acute form and as endocarditis in the chronic form in humans. Its presence in Turkey has been shown with a large number of studies. The aim of this study was to show the genotypic relationship with MLVA analysis of C. burnetii samples found in cattle, sheep and goat samples in Erzurum and Samsun Veterinary Control Institutes and blood samples collected from humans with atypical pneumonia findings. In the study, MLVA analyses of 100 positive samples from 50 cows, 41 sheep and 9 goats from Northeast Anatolia and Black Sea regions and C. burnetii positive samples found in 6 individuals with atypical pneumonia were performed. As a result of the study, it was found that 106 C. burnetii samples had belong to 16 genotype groups. It was found that genotype XVI was the most prevalent among these groups and it was seen in both regions. In addition to this, genotype IX profile was the second largest group with 83.3% (5/6) of human samples. In this study, the genotypes common in the regions were determined and a data source was created for possible outbreaks.