Geese as reservoirs of human colon cancer-associated Streptococcus gallolyticus.

Recently, an increased number of reports have described pathogens of animal origin that cause a variety of infections and a rise in their transmission to humans. Streptococcus gallolyticus, a member of the Streptococcus bovis/Streptococcus equinus complex (SBSEC), is one of these pathogens and infects a wide range of hosts from mammals to poultry and has a broad functionality ranging from pathogenicity to food fermentation. As S. gallolyticus causes complications including bacteremia, infective endocarditis, and colorectal malignancy in humans, it is important to investigate its occurrence in various hosts, including geese, to prevent potential zoonotic transmissions. This study aimed to investigate the presence of S. gallolyticus in the droppings of clinically healthy and diarrheic geese, which were raised intensively and semi-intensively, by the in vitro culture method, characterize the isolates recovered by PCR and sequence-based molecular methods and determine their antibiotic susceptibility by the disk diffusion and gradient test methods. For this purpose, 150 samples of fresh goose droppings were used. Culture positivity for S. gallolyticus was determined as 8% (12/150). PCR analysis identified 54.55% (n = 6) of the isolates as S. gallolyticus subsp. gallolyticus and 45.45% (n = 5) as S. gallolyticus subsp. pasteurianus. Following the 16S rRNA sequence and ERIC-PCR analyses, S. gallolyticus subspecies exhibited identical cluster and band profiles that could be easily distinguished from each other and were clonally identified. High rates of susceptibility to florfenicol, penicillin, rifampicin, and vancomycin were detected among the isolates, regardless of the subspecies diversity. Both subspecies showed high levels of resistance to bacitracin, clindamycin, doxycycline, tetracycline, trimethoprim-sulfamethoxazole, and erythromycin and multiple MDR profiles, indicating their potential to become superbugs. This first report from Türkiye demonstrates the occurrence of the S. gallolyticus subspecies in geese. In view of the recent increase of geese production and the consumption of goose meat in Türkiye, the occurrence of S. gallolyticus in geese should not be ignored to prevent zoonotic transmission.

Molecular detection of bacterial zoonotic abortive agents from ruminants in Turkey.

Abortions in cattle and sheep are one of the major causes of economic losses worldwide. Brucella spp. are the most common infectious agent associated with these abortions. However, abortions caused by bacteria such as Listeria spp., Leptospira spp., Campylobacter spp. and Mycoplasma spp. are usually overlooked due to their sporadic nature and their status as non-priority abortion agents. In our study, we investigated the bacteria associated with abortion cases in cattle and sheep using PCR. For this purpose, we collected vaginal swab samples (n: 110) of aborted cattle and sheep, as well as stomach content samples (n: 69) of aborted calves and lambs from various cities in Turkey. The samples were analysed by bacteria-specific PCR to detect Campylobacter fetus, Leptospira spp., Listeria spp., Mycoplasma spp., and Yersinia spp. PCR analyses revealed that the investigated bacterial agents were present in 18.85% and 19.3% of the cattle and sheep samples, respectively, with an overall percentage of 18.99%. While the overall positivity rate for C. fetus, Leptospira spp., and Mycoplasma spp. was 2.79%, 10.06%, and 4.47%, respectively, the positivity rate for co-infection with Leptospira spp. and C. fetus was 1.68%. All samples were found to be negative for Yersinia spp. and Listeria spp. The high C. fetus positivity rate detected in sheep and in the stomach contents was statistically significant (p < 0.05). However, the difference in positivity rates between the cities, hosts, co-infections and causative agents was statistically insignificant (p > 0.05). This study provides preliminary data on the significant involvement of C. fetus, Leptospira spp. and Mycoplasma spp. in cattle and sheep abortions in Turkey indicating that they should not be overlooked in diagnosis. In addition, further research is needed to investigate the zoonotic potential of these pathogens for public health in Turkey.

Surface defects due to bacterial residue on shrimp shell.

The changes in the surface chemistry and morphological structure of chitin forms obtained from shrimp shells (ShpS) with and without microorganisms were evaluated. Total mesophilic aerobic bacteria (TMAB), estimated Pseudomonas spp. and Enterococcus spp. were counted in Shp-S by classical cultural counting on agar medium, where the counts were 6.56 ± 0.09, 6.30 ± 0.12, and 3.15 ± 0.03 CFU/g, respectively. Fourier Transform Infrared (FTIR) Spectroscopy and Scanning Electron Microscopy (SEM)/Energy dispersed X-ray (EDX) were used to assess the surface chemistry/functional groups and morphological structure for ChTfree (non-microorganism), and ChTmo (with microorganisms). ChTfree FTIR spectra presented a detailed chitin structure by OH, NH, and CO stretching vibrations, whereas specific peaks of chitin could not be detected in ChTmo. Major differences were also found in SEM analysis for ChTfree and ChTmo. ChTfree had a flat, prominent micropore, partially homogeneous structure, while ChTmo had a layered, heterogeneous, complex dense fibrous, and lost pores form. The degree of deacetylation was calculated for ChTfree and ChTmo according to FTIR and EDX data. The results suggest that the degree of deacetylation decreases in the presence of microorganisms, affecting the production of beneficial components negatively. The findings were also supported by the molecular docking model.

Human Exposure to Naturally Occurring Bacillus anthracis in the Kars Region of Eastern Türkiye.

Environmental contamination with Bacillus anthracis spores poses uncertain threats to human health. We undertook a study to determine whether inhabitants of the anthrax-endemic region of Kars in eastern Türkiye could develop immune responses to anthrax toxins without recognised clinical infection. We measured anti-PA and anti-LF IgG antibody concentrations by ELISA in serum from 279 volunteers, 105 of whom had previously diagnosed anthrax infection (100 cutaneous, 5 gastrointestinal). Of the 174 without history of infection, 72 had prior contact with anthrax-contaminated material. Individuals were classified according to demographic parameters, daily working environment, and residence type. All villages in this study had recorded previous animal or human anthrax cases. Stepwise regression analyses showed that prior clinical infection correlated strongly with concentrations at the upper end of the ranges observed for both antibodies. For anti-PA, being a butcher and duration of continuous exposure risk correlated with high concentrations, while being a veterinarian or shepherd, time since infection, and town residence correlated with low concentrations. For anti-LF, village residence correlated with high concentrations, while infection limited to fingers or thumbs correlated with low concentrations. Linear discriminant analysis identified antibody concentration profiles associated with known prior infection. Profiles least typical of prior infection were observed in urban dwellers with known previous infection and in veterinarians without history of infection. Four individuals without history of infection (two butchers, two rural dwellers) had profiles suggesting unrecognised prior infection. Healthy humans therefore appear able to tolerate low-level exposure to environmental B. anthracis spores without ill effect, but it remains to be determined whether this exposure is protective. These findings have implications for authorities tasked with reducing the risk posed to human health by spore-contaminated materials and environments.

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.

Tracking the footsteps of Francisella tularensis: Bacteriological and serological monitoring in epidemic areas in Ankara.

The study aimed to detect Francisella tularensis (F. tularensis) in water samples and to investigate the seroreactivity of sheep to tularemia in endemic areas where human tularemia cases have been reported in Ankara, Turkey. For the isolation of F. tularensis, 50 water samples were collected from rural areas of 5 regions of Ankara (Turkey) and selectively cultured on Francis medium supplemented with 8-9 % sheep blood and antibiotics (100 IU/ml penicillin G, 100 mg/L cycloheximide, 80,000 U/L polymixin B). No F. tularensis isolate was cultivated from the water samples. To determine the seroreactivity of sheep to tularemia, 1006 sheep blood samples were collected from the regions, where human tularemia is endemic. A microagglutination test (MAT) identified significant antibody titers, ranging from 1/20-1/640 in 181 (17.99 %) of the investigated sheep sera. Further investigation is required in order to evaluate and confirm a possible epidemiologic relationship between human outbreaks and probable role of sheep or other sources.

Tularemia: a re-emerging tick-borne infectious disease.

Tularemia is a bacterial disease of humans, wild, and domestic animals. Francisella tularensis, which is a Gram-negative coccobacillus-shaped bacterium, is the causative agent of tularemia. Recently, an increase in the number of human tularemia cases has been noticed in several countries around the world. It has been reported mostly from North America, several Scandinavian countries, and certain Asian countries. The disease spreads through vectors such as mosquitoes, horseflies, deer flies, and ticks. Humans can acquire the disease through direct contact of sick animals, consumption of infected animals, drinking or direct contact of contaminated water, and inhalation of bacteria-loaded aerosols. Low infectious dose, aerosol route of infection, and its ability to induce fatal disease make it a potential agent of biological warfare. Tularemia leads to several clinical forms, such as glandular, ulceroglandular, oculoglandular, oropharyngeal, respiratory, and typhoidal forms. The disease is diagnosed through the use of culture, serology, or molecular methods. Quinolones, tetracyclines, or aminoglycosides are frequently used in the treatment of tularemia. No licensed vaccine is available in the prophylaxis of tularemia and this is need of the time and high-priority research area. This review mostly focuses on general features, importance, current status, and preventive measures of this disease.

Effects of different flour blends with varying protein content and quality on dough and crust properties of "etliekmek", a pizza-like traditional food of Turkey.

Etliekmek, which somewhat resembles pizza in terms of crust and toppings, is a widely consumed traditional food in Turkey. It consists of a sheeted dough topped with a mixture of minced meat and some vegetables. In this study, the effects on dough processing and crust properties of etliekmek of five flour blends with varying protein content and quality were investigated. The blends (Blend-1 through Blend-5) consisted respectively of hard-endosperm Bezostaja and soft-endosperm Gerek-79 wheat flours at the ratios of 100:0, 75:25, 50:50, 25:75 and 0:100. In addition to pysicochemical properties of wheats and their flour blends, dough processing and etliekmek crust properties were measured through the instrumental and sensory approaches. It was determined that protein contents and qualities of the blends decreased from Blend-1 through Blend-5. The dough from Blend-1 was judged to be extremely elastic, which resisted to sheeting due to elastic recovery, whereas the dough from Blend-5 was scored to be somewhat weak and easily extensible. The most suitable dough for the processing of etliekmek crust, i.e., optimally elastic and properly extensible dough with appropriate sheetability, seemed to be 50:50 blend of Bezostaja and Gerek-79 flours (Blend-3). Blend-1 yielded etliekmek crust with thick, moist and excessively chewy texture, as opposed to the thinner, drier and rather crunchy crust texture from Blend-5. Based on the dimensional measurements and sensory evaluations, Blend-3 yielded the best etliekmek crust. The results demonstrate that an optimum balance of dough viscosity and elasticity, which are mostly governed by flour protein content and quality, is of vital importance to the production of high-quality etliekmek crust.

[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.

The identification of novel single nucleotide polymorphisms to assist in mapping the spread of Bacillus anthracis across the Southern Caucasus.

Anthrax is common as a zoonotic disease in the southern Caucasus area including parts of Turkey and Georgia. In this region, population genetics of the etiological agent Bacillus anthracis comprises, where known, the major canonical single nucleotide polymorphism (canSNP) groups A.Br.Aust94 and A.Br.008/009 of the pathogen's global phylogeny, respectively. Previously, isolates of B. anthracis from Turkey have been genotyped predominantly by multi locus variable number of tandem repeat analysis (MLVA) or canSNP typing. While whole genome sequencing is the future gold standard, it is currently still costly. For that reason we were interested in identifying novel SNPs which could assist in further distinguishing closely related isolates using low cost assay platforms. In this study we sequenced the genomes of seven B. anthracis strains collected from the Kars province of Eastern Anatolia in Turkey and discovered new SNPs which allowed us to assign these and other geographically related strains to three novel branches of the major A-branch canSNP-group (A.Br.) Aust94. These new branches were named Kafkas-Geo 1-3 and comprised isolates from the Kars region and the neighboring republic of Georgia suggesting a common ancestry. The novel SNPs identified in this study connect the population genetics of B. anthracis in the South Caucasus and Turkey and will likely assist efforts to map the spread of the pathogen across this region.

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.

Caenorhabditis elegans Predation on Bacillus anthracis: Decontamination of Spore Contaminated Soil with Germinants and Nematodes.

Remediation of Bacillus anthracis-contaminated soil is challenging and approaches to reduce overall spore levels in environmentally contaminated soil or after intentional release of the infectious disease agent in a safe, low-cost manner are needed. B. anthracis spores are highly resistant to biocides, but once germinated they become susceptible to traditional biocides or potentially even natural predators such as nematodes in the soil environment. Here, we describe a two-step approach to reducing B. anthracis spore load in soil during laboratory trials, whereby germinants and Caenorhabditis elegans nematodes are applied concurrently. While the application of germinants reduced B. anthracis spore load by up to four logs depending on soil type, the addition of nematodes achieved a further log reduction in spore count. These laboratory based results suggest that the combined use of nematodes and germinants could represent a promising approach for the remediation of B. anthracis spore contaminated soil. Originality-Significance Statement: This study demonstrates for the first time the successful use of environmentally friendly decontamination methods to inactivate Bacillus anthracis spores in soil using natural predators of the bacterium, nematode worms.

Virulence plasmid stability in environmentally occurring Bacillus anthracis from North East Turkey.

The Bacillus anthracis virulence plasmid pXO2, which encodes for a polypeptide capsule, can be lost during long term laboratory storage. To determine if pXO2 is lost in nature we screened B. anthracis isolates obtained from B. anthracis spores from contaminated animal burial sites in Turkey for their ability to express a capsule upon primary culture. A total of 672 B. anthracis colonies were examined of which ten produced a mixed mucoid (capsule +ve)/non-mucoid (capsule -ve) phenotype and a further one colony yielded non-mucoid colonies upon repeated culture. Screening by PCR using pXO2 specific primers revealed that seven of these isolates had eliminated the plasmid. Of the four colonies which were positive by PCR, one regained the ability to express a capsule upon repeated culture suggesting that the defect was reversible. This is an important observation as capsule expression is a principal marker of virulence and in the absence of PCR serves as a key diagnostic marker. The results of this preliminary study suggest that pXO2 is lost in nature and that further studies are need to determine the mechanisms by which this occurs.

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.

Efficacy of pomades containing different percentages of enilconazole in the treatment of bovine dermatophytosis.

BACKGROUND: Enilconazole is a broad-spectrum topical antimycotic agent used for the management of bovine dermatophytosis. HYPOTHESIS/OBJECTIVES: To investigate the efficacy of pomades containing different concentrations of enilconazole for the treatment of bovine dermatophytosis. METHODS: Dermatophytosis was confirmed in 120 cattle from farm in Gole region of Turkey. Animals were divided into six groups (n = 20 in each). Pomades containing 1%, 2%, 3%, 4% and 5% enilconazole were applied topically to individual lesions in groups I-V, respectively, once a day for 3 days. Group VI animals were used as a control group. Animals were monitored clinically once a week for a two month period. RESULTS: Cows treated with pomades containing 4% and 5% enilconazole recovered; adverse topical reactions occurred in 40% and 55% of animals, respectively. The success rate for cows treated with pomades containing 3% enilconazole was 95% and they recovered with no adverse reactions. Success rates for treatment were 25% and 50% for cows treated with pomades containing 1% and 2% enilconazole, respectively. No improvement was observed in the control group. CONCLUSIONS AND CLINICAL IMPORTANCE: Pomades containing 3% enilconazole are recommended for the treatment of bovine dermatophytosis.

The Use of Germinants to Potentiate the Sensitivity of Bacillus anthracis Spores to Peracetic Acid.

Elimination of Bacillus anthracis spores from the environment is a difficult and costly process due in part to the toxicity of current sporicidal agents. For this reason we investigated the ability of the spore germinants L-alanine (100 mM) and inosine (5 mM) to reduce the concentration of peracetic acid (PAA) required to inactivate B. anthracis spores. While L-alanine significantly enhanced (p = 0.0085) the bactericidal activity of 500 ppm PAA the same was not true for inosine suggesting some form of negative interaction. In contrast the germinant combination proved most effective at 100 ppm PAA (p = 0.0009). To determine if we could achieve similar results in soil we treated soil collected from the burial site of an anthrax infected animal which had been supplemented with spores of the Sterne strain of B. anthracis to increase the level of contamination to 10(4) spores/g. Treatment with germinants followed 1 h later by 5000 ppm PAA eliminated all of the spores. In contrast direct treatment of the animal burial site using this approach delivered using a back pack sprayer had no detectable effect on the level of B. anthracis contamination or on total culturable bacterial numbers over the course of the experiment. It did trigger a significant, but temporary, reduction (p < 0.0001) in the total spore count suggesting that germination had been triggered under real world conditions. In conclusion, we have shown that the application of germinants increase the sensitivity of bacterial spores to PAA. While the results of the single field trial were inconclusive, the study highlighted the potential of this approach and the challenges faced when attempting to perform real world studies on B. anthracis spores contaminated sites.

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.