A comprehensive study of Helicobacter pylori infection: molecular analysis, antibacterial susceptibility, and histopathological examination.

Helicobacter pylori is a pathogen associated with gastroduodenal diseases. This study aimed; (i) to investigate H. pylori presence by invasive tests in adult dyspeptic patients, (ii) to determine antibiotic susceptibility and genotypic characteristics of the H. pylori isolates, and (iii) to investigate the relationship between the H. pylori genotypes and the histopathological findings. In this cross-sectional study, gastric biopsy samples from 208 adult dyspeptic patients were used for culture, tissue Polymerase Chain Reaction (PCR), and histopathological analysis. Antibiotic susceptibility of the H. pylori isolates was analyzed by gradient method. Analysis of the virulence genes was performed by monoplex PCR. Genetic profiles (from A to H) were created based on the virulence genes presence. Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) was used for the genotyping of the H. pylori isolates. The mean age of the patients was 46 (± 15) years and 128 (61.5%) of them were female. H. pylori positivity was detected by culture, tissue PCR and histopathological examination in 59 (28.4%), 114 (54.8%) and 81 (38.9%) patients, respectively. The overall prevalence of H. pylori was found to be 63% (131/208). All H. pylori isolates were susceptible to tetracycline and amoxicillin. The resistance rates for metronidazole, clarithromycin, levofloxacin, and rifampicin were 67.2%, 27.9%, 34.4% and 13.11%, respectively. Multi drug resistance (MDR) was detected at the rate of 45.9% (28/61). While the most common virulence gene was cagA (93.44%), the least common was vacAm1 (23%). The predominant genetic profile was profile A (47.5%). ERIC-PCR results revealed a total of 26 different patterns. A high prevalence of H. pylori was detected in adult dyspeptic patients as in developing countries. It was observed significant genotypic heterogeneity and virulence gene diversity within the isolates. A considerable resistance rate detected against antibiotics such as clarithromycin, metronidazole, and levofloxacin, which are frequently used in the eradication of H. pylori, should be taken into consideration when creating regional empirical treatment regimens.

The canonical Brucella species-host dependency is changing, however, the antibiotic susceptibility profiles remain unchanged.

Brucellosis is a chronic disease caused by Brucella species with a wide range of hosts, from marine mammals to terrestrial species, but with strict host preferences. With the zoonotic character, the prevalence of human brucellosis cases is a reflection of animal infections. This study aimed to identify 192 Brucella isolates obtained from various sources by Bruce-ladder PCR and to determine their antibiotic susceptibilities by gradient diffusion method (E-test). As a result of the PCR, all human isolates (n = 57) were identified as B. melitensis. While 58 (82.9%) of the cattle isolates were identified as B. abortus, 59 (90.8%) of the sheep isolates were identified as B. melitensis. In addition, 12 (17.1%) of the cattle isolates and 6 (9.2%) of the sheep isolates were determined as B. melitensis and B. abortus, respectively. The primary host change behavior of B. melitensis was 1.9 times higher than that of B. abortus. While gentamicin and ciprofloxacin susceptibilities of Brucella isolates were 100%, tetracycline, doxycycline, streptomycin, trimethoprim/sulfamethoxazole and rifampicin susceptibilities were 99%, 99%, 97.4%, 91.7% and 83.9%, respectively. The lowest sensitivity of the isolates was determined against to cefoperazone as 26%. A triple-drug resistance was detected in 1 B. abortus isolate that included simultaneous resistance to cefoperazone, rifampicin, and trimethoprim/sulfamethoxazole. The high susceptibility profiles we found against to antibiotics such as tetracycline, doxycycline gentamicin and ciprofloxacin, used widely in treatment, are encouraging. However, the change in the canonical Brucella species-primary host preference suggests the need to reconsider eradication program, including updating vaccine formulations.

[Helicobacter pylori Positivity and Risk Analysis in Patients with Abdominal Pain Complaints]. / Abdominal Agri Yakinmali Hastalarda Helicobacter pylori Pozitifligi ve Risk Analizi.

Helicobacters have wide host diversity due to the their particular virulence and environmental factors and may cause infections in humans. As they live in and around the stomach the group is called as gastric helicobacters which particularly consists of Helicobacter pylori and Helicobacter heilmanni, Helicobacter felis, Helicobacter salomonis and many other species, as well. In this study, it was aimed to evaluate 195 patients (119 urban and 76 rural residents, 121 female and 74 male individuals between 18 and 93 years of age) in terms of gastric Helicobacter (H.pylori, H.felis and H.heilmanii) who have admitted to the Health Research and Application Center of Kafkas University Endoscopy Unit of the General Surgery Department with the complaints of abdominal pain. For this purpose, biopsy specimens obtained from various parts of the stomach (corpus and antrum) by endoscopy were analyzed with histopathological examination and PCR. Histopathological analysis sections were stained with May-Grunwald-Giemsa and spiral-shaped helicobacters attached to the surface of the epithelium were investigated. For the direct analysis of Helicobacter in biopsy samples, 16S rRNA gene based genus-specific and urease B gene based species-specific PCR methods were used. Out of the 195 cases that were histopathologically evaluated 163 (83.58%) were found to be positive for gastric Helicobacter, while five were suspected and 27 were negative. Helicobacter spp. DNA were detected in 107 (54.87%) samples, of these samples 91 were histopathologically positive, 13 were negative and three were suspicious samples. Eighty seven (44.61%) of the samples were identified as H.pylori by species-specific PCR. H.felis and H.heilmannii could not be detected in any of the samples; meanwhile genus-specific PCR positive 20 samples were not identified. In this study, 42.85% of the individuals living in urban area and 47.36% of those living in rural area were identified as H.pylori positive. 46.28% of women and 41.89% of men were positive for H.pylori. The age range of H.pylori positive individuals were as follows: 60% of the individuals were between 15-24 years, 60.27% of the individuals were between 25-44 years, 34.66% of the individuals were between 45-64 years and 29.72% of the individuals were 65 and over. 42.64% of the cat or dog owners were found as H.pylori positive whereas H.pylori was positive in 45.66% of the individuals who do not own animals. No significant relationship was found between these determinants and the prevalence of the disease (p> 0.05). However, the positivity of H.pylori was higher in the 25-44 active working age group due to the increased agent exposure (p<0.05). This study is the first study on the prevalence of H.pylori in humans and analysis of possible risk factors in the region and hoped to provide useful information for the researchers working in this field.

Protection of farm goats by combinations of recombinant peptides and formalin inactivated spores from a lethal Bacillus anthracis challenge under field conditions.

BACKGROUND: Bacillus (B.) anthracis, the causal agent of anthrax, is effectively controlled by the Sterne live spore vaccine (34F2) in animals. However, live spore vaccines are not suitable for simultaneous vaccination and antibiotic treatment of animals being at risk of infection in an outbreak situation. Non-living vaccines could close this gap. RESULTS: In this study a combination of recombinant protective antigen and recombinant Bacillus collagen-like antigen (rBclA) with or without formalin inactivated spores (FIS), targeted at raising an immune response against both the toxins and the spore of B. anthracis, was tested for immunogenicity and protectiveness in goats. Two groups of goats received from local farmers of the Kars region of Turkey were immunized thrice in three weeks intervals and challenged together with non-vaccinated controls with virulent B. anthracis, four weeks after last immunization. In spite of low or none measurable toxin neutralizing antibodies and a surprisingly low immune response to the rBclA, 80% of the goats receiving the complete vaccine were protected against a lethal challenge. Moreover, the course of antibody responses indicates that a two-step vaccination schedule could be sufficient for protection. CONCLUSION: The combination of recombinant protein antigens and FIS induces a protective immune response in goats. The non-living nature of this vaccine would allow for a concomitant antibiotic treatment and vaccination procedure. Further studies should clarify how this vaccine candidate performs in a post infection scenario controlled by antibiotics.

The role of staphylococci in subclinical mastitis of cows and lytic phage isolation against to Staphylococcus aureus.

AIM: This study was conducted to determine the role of Staphylococcus in the formation of subclinical mastitis in cows and to isolate the phage against isolated Staphylococcus aureus strains. MATERIALS AND METHODS: In this study, 400 milk cows were screened by California Mastitis Test (CMT) for subclinical mastitis and 235 udders of 96 cows, which were determined to be positive, were evaluated for Staphylococcus. Milk samples were evaluated using conventional and molecular methods. In addition, phage isolation studies were performed against S. aureus strains causing mastitis. RESULTS: At the result of cultural examination, of 235 milk samples that were found as positive for mastitis by CMT, a total of 117 (49.7%) Staphylococcus spp. were isolated as a distribution of 74 (63.24%) coagulase-positive staphylococci and 43 (36.75%) coagulase-negative staphylococci. Of these isolates, 76 (64.95%) were characterized as S. aureus both conventional and molecular techniques. Lytic bacteriophages against two S. aureus strains which were isolated from mastitic milk samples were obtained from wastewater samples. CONCLUSION: The results of this study show that a significant portion of subclinical mastitis was formed by staphylococci. In addition, phage isolation against S. aureus strains isolated can be considered as one of the steps to be applied in the prophylaxis and treatment of such infections.

The prevalence of tularemia in occupational groups that have contact with animals.

BACKGROUND/AIM: The aim of the current study was to investigate the presence of antibodies against Francisella tularensis in individuals in different occupations that have contact with animals in the Kars region of northeastern Turkey. MATERIALS AND METHODS: A total of 201 blood samples specifically including 103 farmers, 45 clinical veterinarians, 42 butchers, and 11 hunters were analyzed. The results of the study were reported in relation to some sociodemographic features (age, sex, occupation, and experience) of the volunteers. The presence of antibodies was determined by a microagglutination (MA) test. In addition, positive sera were confirmed using an ELISA kit. RESULTS: Fifteen (7.46%) individuals, including fourteen farmers and one clinical veterinarian, were found to be positive for F. tularensis by both MA and ELISA with a titer range of 1/10 to 1/160. The highest seroprevalence rate was observed in farmers (13.59%), followed by clinical veterinarians (2.22%). The occurrence of tularemia was found to increase with age. CONCLUSION: Though the main route of tularemia outbreaks is water-borne in Turkey, it was determined that people whose occupations bring them into contact with animals are at risk. Similar studies are recommended in order to further clarify the epidemiology of the disease in the northeast of Turkey.

The effect of prolonged storage on the virulence of isolates of Bacillus anthracis obtained from environmental and animal sources in the Kars Region of Turkey.

The stability of the plasmid-mediated virulence factors of Bacillus anthracis, a tripartite toxin located on pXO1 and an antiphagocytic capsule encoded by genes located on pXO2, following long-term storage was investigated. A collection of 159 isolates of B. anthracis were collected from the Kars region of Turkey between 2000 and 2013 and stored at -20°C in Brucella broth supplemented with 20% glycerine. A total of 142 isolates were recovered of which one failed to express a capsule upon primary culture. A further 35 isolates yielded a mixture of mucoid and non-mucoid colonies; the majority of which had lost the pXO2 plasmid as determined by PCR analysis. Results would suggest that pXO2 is more unstable than pXO1 and that this instability increases with the length of storage. It is possible that the pXO2-deficient isolates of B. anthracis described here could be developed into a vaccine to treat at risk animals in the Kars region as many animal vaccines are based upon pXO2 deficiency.

Investigation of bovine brucellosis in the Northeastern Turkey.

Bovine brucellosis, caused by Brucella abortus, is a significant problem for both public and animal health in Turkey. This study was conducted on the calving seasons between 2001 and 2006. A total of 626 serum samples of cattle obtained from 27 herds with a history of abortions was examined for Brucella antibodies by RBPT, SAT and ELISA. Of the cattle sera analysed, 221 (35.30%) and 206 (32.92%) and 247 (39.45%) were found to be positive by RBPT, SAT and ELISA, respectively. B. abortus was isolated from 48 (32.21%) of 149 lung samples and stomach contents of the aborted fetuses. Based on the biochemical tests and the agglutination tests with monospecific A and M antisera, only 3 of the isolates were found to be B. abortus biotype 1 and the remaining 45 were biotype 3. This study also revealed that the dominant biotype of B. abortus was biotype 3 in this region. The determination of the agents responsible for bovine brucellosis and serosurvey of this disease are expected to help better understanding of this zoonotic infection in this region and neighbouring countries.

Isolation of various Arcobacter species from domestic geese (Anser anser).

In this study, the prevalence and distribution of various Arcobacter spp. were investigated in samples taken from the cloacae of healthy domestic geese raised in Turkey. A membrane filtration technique with a non-selective blood agar was employed after enrichment in Arcobacter enrichment broth (AEB) to isolate a wide range of Arcobacter spp. In addition, the isolates were characterized phenotypically and identified at species level using a multiplex-PCR assay. A total of 90 cloacal swab samples taken from geese, collected on three farms (18, 25, 47 samples, respectively), were examined. Of the samples examined, 16 (18%) were found positive for Arcobacter. One Arcobacter species was isolated from each bird. Of the 16 Arcobacter isolates, 7 (44%), 7 (44%) and 2 (12.5%) were identified by m-PCR as A. cryaerophilus, A. skirrowii and A. butzleri, respectively. The present study indicates that domestic geese can harbour a variety of Arcobacter spp. in their cloacae. The presence of Arcobacter in geese may be of significance as reservoirs in their dissemination. Detailed research is needed for better understanding of the epidemiology and zoonotic potential of this emerging pathogen.

Outbreaks of tularemia in Turkey.

Tularemia, casued by Francisella tularensis, is a zoonotic disease presenting various clinical forms. In the present study, three outbreaks of tularemia occurred from January to March and September in 2004 (first and second) and January to March in 2005 (third) are reported from the north-eastern part of Turkey. All cases originated from the same geographical location. In total, 56 patients having complaints of fever, malaise, chills and shivering, painful sore throat with swollen tonsils and enlarged cervical lymph nodes were affected and the patients were different in all cases. Forty-four, 7 and 5 people were affected in the first, second and third outbreak, respectively. The sera from all patients were analysed for the presence of F. tularensis antibodies using a microagglutination assay. Overall, of the 56 sera analysed, 39 (33, 3 and 3 were from the first, second and third outbreak, respectively) showed antibody titres of 1/160 and/or more against F. tularensis. The current report suggests that tularemia exists in north-eastern part of Turkey. The clinical manifestation of the current cases were similar to those of oropharyngeal form of tularemia. It is considered that this region should be accepted as an endemic area for tularemia and kept under control for a long period.