Chitinolytic bacteria in the minke whale forestomach

Minke whales consume large amounts of pelagic crustaceans. Digestion of the prey is initiated by indigenous bacteria in a rumen-like forestomach system. A major structural component of the crustacean exoskeleton is chitin, the beta-1,4-linked polymer of N-acetyl-D-glucosamine. The exoskeletons appear to dissolve completely in the non-glandular forestomach. Bacteria in the forestomach fluid of six krill-eating minke whales were enumerated and isolated using an anaerobic habitat-simulating culture medium. Median viable population densities ranged between 6.0 x 10(6) and 9.9 x 10(9) bacterial cells per mL forestomach fluid. Bacterial isolates (n = 44) cultured from the forestomach fluid of one minke whale mainly resembled strains of Eubacterium (25%), Streptococcus (18%), Clostridium (14%), and Bacteroides (11%). As much as 12% of the bacterial isolates were chitinolytic, while beta-N-acetylglucosaminidase activity was demonstrated in 54% of the isolates, and utilisation of N-acetyl-D-glucosamine was observed in 73%. The chitinolytic isolates resembled strains of Bacteroides, Bacteroidaceae, Clostridium, and Streptococcus. Scanning and transmission electron microscopy of partly digested krill from the minke whale forestomach revealed bacteria close to and inside the chitinous exoskeleton. The bacterial chitinase may act on the chitinous crustacean exoskeletons, thereby allowing other bacteria access to the nutritious soft inner tissues of the prey, and thus initiating its degradation and fermentation.     

Fishing gears involved in entanglements of minke whales (Balaenoptera acutorostrata) in the East Sea of Korea

Entanglement of marine mammals in fishing gear is a global issue. It is considered a significant threat to minke whales (Balaenoptera acutorostrata) in the East Sea of Korea. A total of 214 entanglements of minke whales in this area between 2004 and 2007 were used to investigate types and parts of fishing gears involved in entanglements. The majority of entanglements were mainly caused by three types of fishing gears: set nets, pots, and gill nets (n= 207, 96.7%). Other entanglements were associated with bottom trawls, purse seines, and trawls. A total of 65 entanglements were attributed to the main and branch lines of fishing gears. The most common body part of minke whales which attached to fishing gears was the mouth (n= 63, 30.4%). Most entanglements took place within 10 nmi from land (n= 179, 86.5%), and between 10 and 220 m of water depth. The mean length of entangled minke whales in set nets was significantly smaller than that of whales in pots and gill nets samples (P < 0.001). Also, the mean body length of minke whales that entangled in the coastal area and shallow waters was significantly shorter than that of whales in the offshore area and deep waters (P < 0.001). This information can be used as fundamental data to conserve and manage this population of minke whales in the East Sea of Korea, and also to modify fishing gear to reduce entanglements. Future studies should focus on investigating the impact of these entanglements on the population and the effectiveness of mitigation measures to reduce entanglements of minke whales in this area.     

Anaerobic bacteria from the digestive tract of North Atlantic fin whales (Balaenoptera physalus)

Abstract Samples were collected from the forestomach and colon of North Atlantic fin whales ( Balaenoptera physalus ) landed at the commercial whaling station at Hvalfjördur, Iceland during three whaling seasons. Techniques were used to enrich for and enumerate anaerobic bacteria, methanogens, and sulfate reducers. Anaerobic bacteria ranged from 10⁸ to 10¹⁰ per ml of digesta in the colon, and from 10⁵ to 10⁹ per ml of digesta in the forestomach. Methanogens and sulfate-reducing bacteria were found in the majority of forestomach and colon samples, with sulfate-reducing bacteria usually occuring at higher concentrations. Enteric bacteria, Vibrio , and Listonella spp. were found in the colon. Volatile fatty acids were detected in significant concentrations in the forestomach of many of the whales. These results support previous findings which suggest that a microbial fermentation occurs in the forestomach of baleen whales.     

Anisakis spp. in toothed and baleen whales from Japanese waters with notes on their potential role as biological tags

In this study, Anisakis nematodes isolated from toothed and baleen whales from localities around Japan were molecularly (PCR-RFLP) identified. In Wakayama, common bottlenose dolphins (Tursiops truncatus) were infected with A. simplex sensu stricto (s.s.), A. typica and A. pegreffii, while A. typica was the only species found in pantropical spotted dolphin (Stenella attenuata) and striped dolphin (S. coeruleoalba). Offshore common minke whales (Balaenoptera acutorostrata) and sei whales (B. borealis) were almost exclusively infected with A. simplex s.s.. However, in common minke whales from two Hokkaido localities, mature worms mostly consisted of A. simplex s.s. in some individuals and of A. pegreffii in others, but immature worms were mainly A. simplex s.s.. Gross and histopathological examination on gastric mucosa attached by anisakids resulted in mild and superficial reactions by the two baleen whale species in contrast to severe inflammatory reaction associated with ulcer formations by common bottlenose dolphin. Host specificity and adaptability of Anisakis spp. in these baleen and toothed whales were discussed from the points of view of adult worm size, worm population and pathological reactions by hosts. Interestingly, most of the common minke whales predominantly harboring mature A. pegreffii adults belonged to the Yellow Sea – East China Sea stock (J stock), which migrates through the Sea of Japan, whereas most of those mainly parasitized by mature A. simplex s.s. adults were from the Okhotsk Sea – West Pacific stock (O stock), mostly inhabiting the Pacific side, suggesting that these sibling species may have utility as biological tags to differentiate whale stocks. These results represent the first definitive host records for A. pegreffi in the Northwestern Pacific Ocean.     

Cultivable internal bacterial flora of ticks isolated in Hungary

From six sampling sites in north-western Hungary 126 questing ticks of three species (Ixodes ricinus, Dermacentor reticulatus, Haemaphysalis concinna) were sampled. After inactivating the external bacteria on the surface of ticks, the internal bacterial flora was cultured (two kinds of agar media at three temperatures with aerobic and anaerobic conditions were applied), and 116 strains were isolated. Our results showed, that after a blood meal the bacterial contamination of ticks was much higher in Ixodes and Dermacentor, than in Haemaphysalis, indicating different host range or blood meal habits. Most (89.7 %) of the bacteria were Gram-positive, the most frequent genera were the Staphylococcus (18.1 %) and Bacillus (7.8 %). The percentage of bacteria part of the skin and mucosal flora was 21.6 %. Among the environmental bacteria 14 were found with reported medical importance. The results show, that members of some genera are able to replicate inside the ticks (Mycobacterium, Bacillus) which can increase their potential risk. Isolated bacteria/tick ratio continuously grew from larvae to adults, indicating that larvae probably are hatched sterile, but later bacterial uptake from the environment and from the hosts increases bacterial contamination. Ten anaerobic bacteria were cultured, mostly Propionibacterium acnes, a facultative skin pathogen. No significant differences were found between the isolated bacteria of the six sampling sites. Our work showed, that internal bacterial community of ticks is diverse, novel strains were isolated several with medical importance, some bacteria multiplicate inside ticks, and that ticks continuously take up bacteria from the environment. Our study first described anaerobic bacteria from ticks.     

Identification of Novel Helicobacter spp. from a Beluga Whale

The gastric fluid and feces of three belugas from the Mystic Aquarium were assessed for the presence of Helicobacter spp. Gastric fluid and feces from the two clinically healthy belugas were negative for helicobacter, and endoscopy performed on these animals revealed no lesions. However, a helicobacter isolate and PCR product similar to helicobacter strains previously recovered from dolphins were identified, respectively, from the feces and gastric fluid of a beluga manifesting intermittent inappetence and lethargy. Esophageal and forestomach ulcers were noted on endoscopy. This is the first report of novel Helicobacter spp. being identified from whales.     

The presence of bacteria and yeast like fungi in specimens from surgical patients

The aim of this study was to determine the incidence of Candida spp. strains in specimens obtained from surgically treated patients as well as to analyze the accompanying bacterial flora, both aerobic and anaerobic. The material came from two groups of patients. In the first group consisting of patients operated for colon and rectum carcinoma, the samples included peritoneal fluid, colon or rectum bioptates, pus, blood, and wound swabs. In the other group, biopsy material and smears from post operation wounds were taken from patients who underwent a surgical treatment of larynx carcinoma. Altogether, 282 various clinical specimens from 165 patients were analysed, and 41 Candida spp. strains were isolated: 39 strains of C. albicans and 2 strains of C. tropicalis. In 20 out of 41 specimens infected with Candida spp. (48.8%) the co-infection with bacterial aerobic flora was found. In 10 cases (24.4%), the fungi were isolated together with aerobic and anaerobic bacterial flora, whereas in 2 specimens (4.9%) the anaerobes and Candida albicans were diagnosed. The remaining 9 samples showed only the presence of Candida spp. (21.9%). From among aerobic bacterial flora Enterococcus spp. strains (n = 17) and Gram negative rods from Enterobacteriaceae family (n = 13) were the most frequently isolated. The bacterial strains of Streptococcus spp. (n = 5), Pseudomonas spp. (n = 3), Staphylococcus spp. and Corynebacterium spp. (2 strains, both) were identified more rarely. Bacteroides spp. were the most frequent members of bacterial anaerobic flora (n = 10). Other isolated anaerobic bacteria were classified as Fusobacterium spp. or Peptostreptococcus spp. (1 strain each). E. coli and Enterococcus spp. strains of aerobic bacterial flora were more frequently isolated together with Candida spp. CONCLUSIONS: (i) Mixed bacterial flora was found to predominate in the clinical material from the patients after surgery. (ii) Candida spp. were most frequently found together with aerobic bacterial flora.     

Diverse bacteria isolated from microtherm oil-production water

In total, 435 pure bacterial strains were isolated from microtherm oil-production water from the Karamay Oilfield, Xinjiang, China, by using four media: oil-production water medium (Cai medium), oil-production water supplemented with mineral salt medium (CW medium), oil-production water supplemented with yeast extract medium (CY medium), and blood agar medium (X medium). The bacterial isolates were affiliated with 61 phylogenetic groups that belong to 32 genera in the phyla Actinobacteria, Firmicutes, and Proteobacteria. Except for the Rhizobium, Dietzia, and Pseudomonas strains that were isolated using all the four media, using different media led to the isolation of bacteria with different functions. Similarly, nonheme diiron alkane monooxygenase genes (alkB/alkM) also clustered according to the isolation medium. Among the bacterial strains, more than 24 % of the isolates could use n-hexadecane as the sole carbon source for growth. For the first time, the alkane-degrading ability and alkB/alkM were detected in Rhizobium, Rhodobacter, Trichococcus, Micrococcus, Enterococcus, and Bavariicoccus strains, and the alkM gene was detected in Firmicutes strains.     

Characterization of the beneficial properties of lactobacilli isolated from bullfrog (<Emphasis Type="Italic">Rana catesbeiana</Emphasis>) hatchery

The present work addresses the isolation and partial identification of the microbial population of a R. catesbeiana hatchery in spring and summer as well as some beneficial properties of Lactobacillus strains isolated in different seasons and hatchery areas. The bacterial population was grouped into the following taxa: Lactobacillus spp., Pediococcus spp., Enterococcus faecalis and Ent. faecium, and Enterobacteriaceae (Enterobacter spp., Escherichia coli) while Pseudomonas aeruginosa and Staphylococcus epidermidis were isolated from frogs displaying red-leg syndrome. The Lactobacillus plantarum and L. curvatus strains isolated showed to inhibit the growth of red-leg syndrome associated pathogens and food-borne bacteria by organic acids. While L. plantarum CRL 1606 also inhibited red-leg syndrome related pathogens by hydrogen peroxide, meat spoilage bacteria were only inhibited by acidity. However, by using a MRS medium added with tetramethyl-benzidine and peroxidase, a high percentage of H₂O₂-producing lactobacilli were detected. The surface properties of Lactobacillus strains showed that a few strains were able to agglutinate ABO human erythrocytes, while the highest number of strains had a low to medium degree of hydrophobicity. This paper constitute the first study related to the beneficial properties of Lactobacillus isolated from a bullfrog hatchery, as well as the selection criteria applied to a group of strains, which could help to control or prevent bacterial infectious diseases in raniculture.     

Isolation and identification of the oral bacteria and their characterization for bacteriocin production in the oral cavity

Oral cavity is a diverse ecosystem which harbors immense diversity of microorganisms like fungi, virus and bacteria. Some of these microorganisms are involved in causing multiple infections. Oral flora is continuously changing due to connection with the external environment and produce bacteriocin against each other to compete for nutrient in this mini ecosystem. Current study was aimed to explore and compare the bacterial fauna of both healthy and non-healthy dental samples, by isolation and identification with biochemical tests to characterize the bacteriocin production. During study 120 swabs were taken from both healthy and unhealthy subjects. Samples were collected from the dental clinics of Makkah City, in sterile eppendorfs containing 1 ml nutrient broth, and were incubated overnight using shaking incubator. Bacteria were isolated following identification through Gram staining, microscopy and biochemical test. Total 15 strains of bacteria were isolated during the study amongst which 8 strains were gram positive and 7 strains were gram negative. The most dominant species of the gram positive strains was Streptococcus pneumoniae (n = 26). On the other hand, Escherichia coli (n = 26) was the prominent specie amongst the gram negative strains. Overall, the dominated family was Enterobacteriaceae (19.36%) followed by Streptococcaceae with 13.83% abundance. One of the most cariogenic strain Klebsiella pneumoniae (n = 14) was also isolated. The bacterial strain diversity between these two type of ecosystem was approximately the same, with slight variation in Shannon (HS:2.627187, NHS:2.653594) and Simpson diversity (HS:0.923461, NHS: 0.92684) index. The current research revealed that bacteriocin production in the Enterobacter species was prominent against Escherichia coli and Klebsiella pneumoniae. Apart from this other strains like Klebsiella pneumoniae and Exiguobacterium spp were also able to produce bacteriocin against Enterobacter species and Bacillus cereus respectively.     

Isolation of new anaerobic,thermophilic and cellulolytic bacteria-JT strains and their cellulase production

Six microbial strains (JT) of endospore-forming, anaerobic, thermophilic and cellulolytic bacteria were isolated from camel feces, compost, soil and hot spring water in Japan. These strains are gram negative and classified as the genus Clostridium. Strains JT3-1, JT3-2 and JT3-3 can digest starch. All of the strains produce a high activity of extracellular cellulases in cellobiose and cellulose media.Strain JT1 produced 1.36 units/ml of CMCase (endo-β-1,4-d-glucanase, EC 3.2.1.4), 66.2 units/ml of β-glucosidase (ED 3.2.1.21) and 39.9 units/ml of β-xylosidase (EC 3.2.1.37) in 1% cellobiose medium. Strain JT3-3 produced 1.87 units/ml of CMCase, 166.3 units/ml of β-glucosidase and 23.6 units/ml of β-xylosidase in 1% cellulose medium.     

Population structure and possible migratory links of common minke whales, Balaenoptera acutorostrata, in the Southern Hemisphere

Mitochondrial DNA control region sequences were analyzed to investigate population structure and possible migratory links of common minke whales (Balaenoptera acutorostrata) in two ocean basins: western South Atlantic (WSA) and western South Pacific (WSP). The results of several different phylogenetic estimations consistently grouped all haplotypes but one (n = 1) from these two ocean basins into two separate clades. South and North Atlantic haplotypes were more closely related to each other than either was to haplotypes from the WSP. The interpopulation genetic distance between WSA and WSP whales was similar to that reported between North Pacific and North Atlantic common minke whales (0.0234). The migration rate between the two ocean basins was estimated at near-zero using MDIV. The genetic evidence presented here was consistent with the hypothesis of migratory links among Brazil, Chilean Patagonia and the Antarctic Peninsula, and between low-latitude and Antarctic waters of the WSP. The results suggest multiple populations of common minke whales in the Southern Hemisphere, which may have conservation as well as taxonomic implications. Our single locus results should be corroborated by additional analyses in a larger number of samples and at more genetic markers.     

The role of anaerobic bacteria in chronic suppurative otitis media in children: Implications for medical therapy

This review describes the microbiology, diagnosis and medical management of chronic suppurative otitis media (CSOM) in children highlighting the role of anaerobic bacteria. In studies that employed adequate method for recovery of anaerobic bacteria polymicrobial aerobic and anaerobic flora was isolated from over half of the children with CSOM. The predominant aerobic isolates were Staphylococcus aureus and Pseudomonas aeruginosa and the most frequently isolated anaerobic organisms were Peptostreptococcus, Fusobacterium spp. and pigmented Prevotella and Porphyromonas spp. Several studies illustrated the efficacy of anti-infective agents effective against anaerobic bacteria in the treatment of CSOM. The medical therapy of CSOM should be directed at the eradication of the pathogenic aerobic and anaerobic organisms.     

Distribution and Physiology of Aerobic Bacteria Containing Bacteriochlorophyll a on the East and West Coasts of Australia

Aerobic heterotrophic bacteria containing bacteriochlorophyll were isolated from specimens from a wide variety of marine environments on the west (Shark Bay, Lake Clifton, Lake Heyward, and Perth) and east (near Townsville and Brisbane) coasts of Australia. The bacteria were found in a high proportion (10 to 30%) of the total heterotrophic bacterial strains isolated from marine algae, seagrasses, stromatolites, the epiphytes on stromatolites, seawater, and sands; in some cases they constituted up to 49% of the total. This is much higher than the previous report of 6% from Japan. A high percentage, 13%, was also found in the seawater of Hamelin Pool, at Shark Bay, where the salinity was 66%. The number of these bacteria was generally low in seawater and sands, with a few exceptions. There were no aerobic bacteriochlorophyll-containing bacteria on sponges or corals. The isolated strains were orange or pink, and most had absorption maxima around 800 and 850 to 870 nm, the latter range being the absorption of bacteriochlorophyll a in vivo. The maximum bacteriochlorophyll content was 1 nmol/mg (dry weight) of bacterial cells. Most of the bacteria did not grow phototrophically under anaerobic conditions in a broth medium containing succinate. Cells and cell extracts grown under aerobic conditions had photochemical activities such as reversible photooxidations of the reaction center and cytochrome(s). Some strains showed denitrifying activity. The optimal salinity for bacterial growth varied between strains.     

Development of media to accelerate the isolation of indigo-reducing bacteria,which are difficult to isolate using conventional media

Indigo-reducing bacteria perform natural fermentation in indigo fermentation fluid. Owing to the stochastic nature of the process, the constituent in indigo fermentation fluid differ depending on the prepared batch and fermentation period. To identify new indigo-reducing bacteria, isolation of the bacteria is indispensable. However, isolation of indigo-reducing bacteria is difficult because conventional media are often unsuitable to isolate these slow-growing bacteria that also exist in low numbers. Hydrolysates of polysaccharides and mixtures of plant base constituents are candidates to accelerate the isolation of indigo-reducing bacteria that cannot be isolated using conventional media. In this current study, wheat bran hydrolysate and composted indigo leaves (sukumo) were used as ingredients in the fermentation fluid in the selective medium for indigo-reducing bacteria in anaerobic culture. The results suggested that obligate and oxygen-non-metabolizing facultative anaerobes are difficult to isolate using conventional media, whereas oxygen-metabolizing facultative anaerobes, relatively rapid-growing and major bacterial strains are relatively easy to isolate. Media containing sukumo hydrolysate facilitated the isolation of novel species of Bacillus pseudofirmus-related strains, whereas media containing wheat bran hydrolysate facilitated the isolation of Amphibacillus spp. (including new species). Seven species (including two new species) of indigo-reducing bacteria were isolated using wheat bran hydrolysate-containing media, whereas six species (including three new species) of indigo-reducing bacteria were isolated using media containing both wheat bran and sukumo hydrolysates. These newly developed culture media will facilitate the isolation of unknown bacteria in indigo fermentation and in environments similar to indigo fermentation fluid.     

Bacterial Population Adherent to the Epithelium on the Roo of the Dorsal Rumen of Sheep

By anaerobic procedures, the total number of adherent bacteria was determined on tissue samples obtained from the roof of the dorsal rumen of three sheep. After four washings, 1.91 × 10⁷, 0.34 × 10⁷, and 1.23 × 10⁷ bacteria per cm² were still attached to the rumen epithelium in sheep 1, 2, and 3, respectively. A total of 95 strains of bacteria were isolated from these three samples. Based on morphology, Gram stain, anaerobiosis, motility, and fermentation end products, they were presumptively identified as follows: Butyrivibrio fibrisolvens, 30 strains; atypical Butyrivibrio, 5 strains; Bacteroides ruminicola, 22 strains; Lactobacillus, 1 strain; and unknown Bacteroides species, 37 strains. For sheep 3, washing the rumen epithelium a total of 10 times reduced the adherent bacterial population by 93% (8.4 × 10⁵ bacteria per cm²). Of 30 strains isolated from this sample, 22 were presumptively identified as Butyrivibrio and Bacteroides types. These results suggest that the epithelium on the roof of the dorsal rumen is primarily colonized by two genera of bacteria, Butyrivibrio and Bacteroides. Most Butyrivibrio and Bacteroides ruminicola strains appeared to be similar to previously isolated rumen strains. However, the unknown Bacteroides species differed considerably from the three species of this genus which are commonly isolated from rumen contents.     

Environmental Isolates of Fungi from Aquarium Pools Housing Killer Whales (Orcinus orca)

Systemic mycoses in killer whales (Orcinus orca) are rare diseases, but have been reported. Two killer whales died by fungal infections at the Port of Nagoya Public Aquarium in Japan. In this study, the fungal flora of the pool environment at the aquarium was characterized. Alternaria spp., Aspergillus spp. (A. fumigatus, A. niger, A. versicolor), Fusarium spp. and Penicillium spp. were isolated from the air and the pool surroundings. The other isolates were identified as fungal species non-pathogenic for mammals. However, the species of fungi isolated from the environmental samples in this study were not the same as those isolated from the cases of disease in killer whales previously reported.     

Diversity Dynamics of Marine Bacteria Studied by Immunofluorescent Staining on Membrane Filters

Of 34 strains of marine bacteria isolated on a general seawater medium, 5 were selected for detailed studies of their population dynamics in the plankton. The isolates were characterized as Aeromonas sp., Chromobacterium cf. lividum, Vibrio sp., and two Pseudomonas spp. Specific antibodies were produced by immunization of rabbits, and bacterial cells were stained on black Uni-Pore membrane filters by an indirect immunofluorescent staining procedure. The method proved to be very specific and practical for use in a large-scale field sampling program. Growth of all five isolates was stimulated by high values for net primary production, chlorophyll a, and dissolved organic carbon. Calculation of a diversity index based on specific and total counts is proposed as a way of characterizing the dynamics of organotrophic bacterial populations in the sea.     

Isolation and Presumptive Identification of Adherent Epithelial Bacteria (“Epimural” Bacteria) from the Ovine Rumen Wall

One hundred sixty-one strains of adherent bacteria were isolated under anaerobic conditions from four sites on the rumen epithelial surface of sheep fed hay or a hay-grain ration. Before isolation of bacteria, rumen tissue was washed six times in an anaerobic dilution solution, and viable bacteria suspended in the washings were counted. Calculation indicated that unattached bacteria would have been removed from the tissue by this procedure, but a slow and progressive release of attached bacteria also occurred. Nevertheless, a wide range of characteristic morphological types remained associated with the epithelium as demonstrated by scanning electron microscopy. Most of these types were represented among the isolates. Characterization and presumptive identification of the isolates showed that 95.0% belonged to previously described genera of functionally significant rumen bacteria, including Butyrivibrio sp. (31.1%), Bacteroides sp. (22.4%), Selenomonas ruminantium (9.9%), Succinivibrio dextrinosolvens (8.7%), Streptococcus bovis (8.1%), Propionibacterium sp. (4.3%), Treponema sp. (3.1%), and Eubacterium sp., Lachnospira multiparus, and Ruminococcus flavefaciens (2.5% each). Eight isolates (5.0%) were not identified. L. multiparus was recovered only from hay-fed animals; all other genera were obtained from animals fed either ration. All S. bovis strains and two strains each of Bacteroides sp. and Butyrivibrio sp. were aerotolerant; all other strains were strictly anaerobic. Bacteria representing the gram-positive, facultatively anaerobic flora associated with rumen wall tissue (R. J. Wallace, K.-J. Cheng, D. Dinsdale, and E. R. Ørskov, Nature (London) 279:424-426, 1979) were therefore not recovered by the techniques used; instead a different fraction of the adherent population was isolated. The term “epimural” is proposed to describe the flora associated with the rumen epithelium.     

Proteomics‐based identification of orchid-associated bacteria colonizing the Epipactis albensis,E. helleborine and E. purpurata (Orchidaceae,Neottieae)

Using proteomics-based identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), we conducted the first analysis of the composition of endophytic bacteria isolated from different parts of selected Epipactis species, i.e. the buds, the inflorescences and the central part of the shoots, as well as the rhizomes. We identified aerobic and anaerobic bacteria, including such taxa as Bacillus spp., Clostridium spp., Pseudomonas spp. and Stenotrophomonas spp., which may be considered as promoting plant growth. Because most of the indicated bacteria genera belong to spore-producing taxa (spores allow bacterial symbionts to survive adverse conditions), we suggest that these bacteria species contribute to the adaptation of orchids to the environment. We found clear differences in the microbiome between investigated closely related taxa, i.e., Epipactis albensis, E. helleborine, E. purpurata and E. purpurata f. chlorophylla. Some of the analysed orchid species, i.e. E. albensis and E. purpurata co-occur in habitats, and their bacterial microbiomes differ from each other.