Mycobacterium abscessus (M. a.), which is considered to be the most resistant to antibiotics and most difficult to treat, has been on the rise. Correct identification to the level of the subspecies  and determination of its sensitivity to macrolides, which are the basis of combination therapy, are of principal importance for the management of the disease. We describe five cases of mycobacterioses caused by M. a., where the sequencing of select genes was performed to identify the individual subspecies and antibiotic resistance. The analysis of the rpoB gene showed two isolates each of M. a. abscessus and M. a. massiliense and one isolate of M. a. bolletii. The complete (full length) erm(41) gene responsible for the development of inducible resistance to macrolides was demonstrated in both M. a. abscessus and M. a. bolletii isolates. A partially deleted and non-functional erm(41) gene was demonstrated in M. a. massiliense isolates. The subsequent sequencing of the full length erm(41) gene products showed, however, the mutation (T28→C) in both isolates of M. a. abscessus, causing a loss of the function and preserved sensitivity to macrolides. The antibiotic sensitivity testing confirmed that both the isolates of M. a. abscessus and M. a. massiliense were sensitive to clarithromycin even after prolonged 14-day incubation. The inducible resistance to clarithromycin was maintained only in M. a. bolletii. Thus, the sequence analysis of the erm(41) gene can reliably identify the preservation of sensitivity to macrolides and serve as an important tool in the establishment of therapeutic regimens in cases of infections with M. abscessus.Mycobacterioses are less frequently occurring but serious diseases. In recent years, at a global level, the incidence of mycobacterioses induced by the rapidly growing species  Mycobacterium abscessus (M. a.) is a rapidly growing nontuberculous Mycobacterium (NTM) belonging to a species of serious clinical importance. Since the beginning of the new millennium, an increase in the occurrence of mycobacterioses has been observed globally [M. avium [globally ,2, whichglobally . The incM. avium ,5. FinalM. avium .M. avium complex (MAC) has remained the most common causative agent of mycobacterioses, while the frequency of other NTM species has shown geographic variability [M. kansasii, M. xenopi, and M. malmoense. This is in contrast to the epidemiological data from North America, Asia, and Australia, where M. a. is the second most frequent causative agent of mycobacterioses [A higher prevalence of mycobacterioses has been recorded mainly in North America, Asia, and Australia, ranging from 7.9 to 14.1/100,000 inhabitants . Around terioses .M. kansasii and M. xenopi. Infections caused by M. a. are, however, recorded only scarcely. Thus, during the years 2012–2018, M. a. was diagnosed according to American Thoracic Society (ATS) Criteria in only four (1.32%) patients with mycobacterioses [M. a. [In the Czech Republic, according to the Information System of Bacillary Tuberculosis (ISBT), the situation is similar to other European countries, where MAC members are the most common causative agents of mycobacterioses, followed by terioses . Neverths [M. a. , especiaM. a. has been shown to be even more difficult, as this species is increasingly resistant to antibiotics. M. a. is intrinsically resistant not only to the classical anti-tuberculous drugs, but also to most of the currently available antibiotics. This resistance is attributed predominantly to the impermeable cell envelope, enzymatic deactivation of drugs, and drug efflux. Susceptibility testing should be performed against a number of antibiotics, including clarithromycin, amikacin, tigecycline, imipenem, cefoxitin, clofazimine, doxycycline, ciprofloxacin, moxifloxacin. The final antibiotic selection should be then guided by drug susceptibility testing. Macrolides represent the critical antibiotic in the combinatory therapy of M. abscessus infections, and the resistance against them may lead to therapy failure [M. a., an introductory combination of at least three effective antibiotics is recommended in the strains sensitive to macrolides, and a combination of at least four antibiotics in resistant strains for a period of at least 1–3 months. This intensive phase usually includes oral macrolide, intravenous amikacin, and one or more of the following: intravenous tigecycline, imipenem, or cefoxitin. This should be followed by a long-term therapeutic regimen with 2–3 effective substances for a period of at least 12 months [Treatment of mycobacterioses is generally complicated as it requires the administration of a combination of antibiotics for a period of at least 12 months owing to the conversion of the sputum to cultivation negativity, according to the recommendation of the ATS ,11. Howe2 months . The con2 months .M. a. ssp. abscessus (M. a. abscessus), M. a. ssp. bolletii (M. a. bolletii), and M. a. ssp. massiliense (M. a. massiliense). Infections caused by M. a. abscessus and M. a. bolletii are generally associated with a clinically worse course and a frequent failure of therapy mainly due to their common resistance to macrolides. This resistance can be constitutive or inducible [M. a. occurs rarely and develops on the basis of spontaneous point mutation in the region of the rrl gene with subsequent selection of the strain during therapy with macrolides. Inducible resistance is connected with the presence of the functional erm(41) gene, which codes for the methylase that alters the target point for the macrolides on the ribosome [M. a. abscessus and M. a. bolletii isolates. M. a. massiliense, however, carry a deletion in the erm(41), and their sensitivity to macrolides is preserved, leading to better treatment outcomes [M. a. abscessus isolates carry mutations in the erm(41) gene, leading to its loss of function and preserved sensitivity to macrolides [At present three subspecies of M. a. are known: nducible . Constitribosome ,17. The ribosome ,19. The outcomes . At the crolides .M. a. and further determine its sensitivity to macrolides.Thus, to apply an effective therapeutic regimen in such infections, it is essential to identify the causative agent to the level of the subspecies of M. a. in patients treated at the University Hospital in Hradec Kralove (Czech Republic), their disease and treatment history, and shows the importance of molecular analysis for the subspecies identification and determination of antibiotic sensitivity.This study describes five cases of otherwise rare clinically important detections of M. a. in the sputum was recorded during a periodical monitoring of the principal disease in August 2013. The clinical condition of the patient was evaluated as stable, without progression, and without a need for additional therapy. Nevertheless, subsequent control examinations of the sputum repeatedly demonstrated M. a. The isolate was further determined as M. a. bolletii (Str 1). In May 2014, the clinical function of the lungs deteriorated with a corresponding finding on high resolution computed tomography (HRCT). After excluding other causes of infection and the demonstration of M. a. bolletii in bronchoalveolar lavage fluid (BAL), an introductory 6-week intravenous therapy was initiated using a combination of amikacin, imipenem, and clarithromycin, which was followed by an oral continuing therapy with doxycycline, clarithromycin, and ciprofloxacin  examination revealed that the Str 1, which was originally sensitive to macrolides, was now resistant. Intravenous therapy was initiated again with the combination of imipenem, tigecycline, amikacin, and azithromycin (for immunomodulatory effect) for a period of 3 months. After that, the patient was treated with oral azithromycin and doxycycline, complemented with inhaled amikacin. Partial regression of the largest infiltrates was observed, and the clinical condition of the patient improved. However, the culture positivity for M. a. bolletii persisted during the entire year of 2015. According to the recommendation of the European Cystic Fibrosis Society (ECFS), clofazimine was added in December 2015. Conversion of the sputum to M. a. bolletii cultivation negativity occurred in May 2017, which was partially due to an improvement of the patient’s compliance. Therapy was terminated in the year 2018.A 19-year-old man with CF; the first detection of Aspergillus niger, demonstrated by cultivation, which was treated with amphotericin (Abelcet) and voriconazole (Vfend). In November 2016, the patient was moved from another facility to the University Hospital in Hradec Kralove (Czech Republic) for further therapy. Because signs of respiratory infection were present on admission, samples from the lower respiratory tract  were obtained, and empirical therapy with ceftazidim, levofloxacin and voriconazole was initiated. The samples from the respiratory tract microscopically demonstrated the presence of acid-fast bacilli (AFB), and the presence of M. a. was identified using the direct PCR method. Subsequently, M. a. was also confirmed in the sample by cultivation, and the isolate was further typed as M. a. abscessus (Str 2). The empirical therapy at the beginning was thus changed for a combination of amikacin, tigecycline, and clarithromycin for 3 months. Cefoxitin, which is not commonly available in the Czech Republic, was added later , nosocomial Pseudomonas aeruginosa pneumonia, and continuing Aspergillus niger infection. After release in 2017, oral therapy with clarithromycin and clofazimine was continued. Sputum, initially cultivation positive for M. a., turned negative 2 weeks after the commencement of targeted therapy, and remained such for the subsequent controls. The patient died 3 months after their release from the hospital due to the worsening GvHD.A 33-year-old patient was diagnosed with myelomonocytic leukaemia in October 2013. After allogeneic transplantation in April 2014, serious Graft versus Host Disease (GvHD) of the lungs developed and was treated with corticoids, methotrexate, and rituximab. In August 2016, pulmonary aspergillosis developed in the terrain of the damaged lungs with M. a. was identified by cultivation in his sputum. The isolate was further typed as M. a. massiliense (Str 3). The patient was clinically stable, without signs of infectious complications. X-ray of the lungs did not show any signs of a new pulmonary affection. The criteria of ATS [M. a. has been repeatedly (twice a year) isolated in the samples of sputum. The patient is still clinically stabilized without a need for targeted therapy.A 68-year-old man was regularly monitored on an out-patient basis due to interstitial pulmonary fibrosis since the year 2015. In January 2018, during a screening examination, a of ATS  were notM. a. was isolated from sputum. The isolate was further determined as M. a. massiliense (Str 4). His clinical condition had been long-term stabilized and satisfactory. After a finding of M. a., CT of the lungs was performed, which showed lesions supporting the diagnosis of lung mycobacteriosis (bronchiolitis and bronchiectasis). Intravenous therapy was commenced with a combination of amikacin, tigecycline, imipenem, and azithromycin for a period of 4 weeks, followed by oral therapy with azithromycin and doxycycline, and with inhalatory amikacin . ConverM. a. was isolated from the sputum. The isolate was further identified to be M. a. abscessus (Str 5). This coincided with a worsening of dyspnea during the exertion reported by the patient. The HRCT examination of the lungs revealed a stationary finding of bronchiectases. On repeated detection of the M. a. abscessus, eradication therapy was initiated with a combination of azithromycin, imipenem, amikacin, and tigecycline for a period of 4 weeks, continuing with azithromycin, moxifloxacin, and inhaled amikacin . ConverrpoB gene of the five isolates demonstrated two isolated strains of M. a. abscessus, two isolated strains of M. a. massiliense, and one isolated strain of M. a. bolletii.Sequencing examination of the rrl gene yielded PCR products of the size of 420 bp in all isolates tested .erm(41) gene was demonstrated in three strains  as a resultant PCR product of the size of 673 bp (erm(41) gene is typical for M. a. abscessus and M. a. bolletii [erm(41) gene is typical for M. a. massiliense.The full length f 673 bp . The prebolletii . In two erm(41) gene was subsequently performed in all strains to verify the identification to the subspecies level. Sequencing analysis of the strains Str 1, Str 2, and Str 5 was additionally used to identify known mutations, which are responsible for the loss of the functional erm(41) gene, leading to the maintenance of the sensitivity to macrolides. Such specific mutation has been described for position 28, where a substitution of T for C (T28→C) [erm(41) gene and development of inducible resistance, are designated as T28 sequevar. Isolates with this mutation, i.e., with the non-functional erm(41) gene and preserved sensitivity to macrolides, are described as C28 sequevar. Another such nucleotide change has been described in position 19 (C19→T) [Sequencing of the  (T28→C)  can occu (C19→T) , which iThe C19→T mutation was observed in none of the isolates. These data suggest that only strain Str 1 should exhibit an inducible resistance to macrolides. The alignment of the sequences of all isolates is available in the To confirm the sequence data and generate the overall antibiotic profile of the studied isolates, the examination of susceptibility to selected antibiotics was performed. The in vitro data showed 100% sensitivity of strains to amikacin, cefoxitin, and tigecycline. Strains also exhibited good sensitivity to imipenem, except for the Str 3. On the other hand, all the isolates were resistant to ciprofloxacin, as well as linezolid and moxifloxacin, with the exception of Str 4 and Str 5, respectively. All isolates were susceptible to clarithromycin after 3 days. However, inducible resistance developed in Str 1 during incubation for 14 days .M. a, which are otherwise relatively rare in the Czech Republic and in Europe in general.Infections with atypical mycobacteria represent an increasing problem worldwide. This study described five clinical cases of infections with M. a. was isolated from the samples from the respiratory tract. The presence of NTM in the respiratory tract does not always indicate pathology, as they can colonize the respiratory tract temporarily or permanently without inducing infection [In all five described cases, nfection ,23. Estanfection ,25. In gnfection . The detnfection . Thus, tM. a. is a recognized causative agent of pulmonary mycobacteriosis, and its isolation from samples obtained from the respiratory tract is certainly a reason to increase attention and to monitor the clinical condition of the patient further [M. a. were identified as a colonization without a clinical relevance only in one patient—Case 3 (Str 3). In the other four cases, the criteria for the diagnosis of mycobacteriosis were fulfilled , and one isolate of M. a. bolletii. Although the number of patients in this study is small, the prevalence of the individual subspecies corresponds with published data, where M. a.abscessus is most frequently demonstrated (45–65%), followed by M. a.massiliense (20–55%), and with M. a. bolletii demonstrated only rarely, in 1–18% [The isolates in each case were further identified by DNA sequencing as in 1–18% .M. a. strains obtained in this study were examined in vitro for their susceptibility to the panel of antibiotics suitable for the treatment of mycobacterioses .terioses . The resrrl gene, which is associated with the constitutive resistance genotypic changes. As expected, none of our isolates showed mutation in this gene. These mutations are relatively rare, e.g., in the paper by Mase et al., it was detected in only one isolate out of 14 [rrl mutants between 87 strains of M. a. These were more often isolated from cystic fibrosis patients; however, this finding was, surprisingly, not significantly associated with a previous treatment. In fact, the acquisition of rrl gene mutations during therapy with macrolides has been reported for several NTM species, in particular for M. a. [Although these results indicated the resistance to be inducible, we have nevertheless performed an analysis of the ut of 14 . Moreoveut of 14  describeor M. a. .erm(41) gene is coding for the methylase that is capable of rendering the carrying bacteria inducible resistant to macrolides [M. a.massiliense isolates corresponded to the fact that this subspecies carried only a truncated and thus non-functional erm(41) gene. While, in general, M. a.abscessus carries a functional erm(41) gene, some isolates remain sensitive to macrolides as a result of a loss of the functional erm(41) gene due to point mutations, the frequency of which is described at about 15–20%. Two kinds of such loss-of-function mutations have been described so far. Thus, T28C mutation leads to the substitution of tryptophan for arginine in codon 10, and such strains are called C28 sequevars. Another mutation C19T introduces a premature stop codon (T19 sequevar) [M. a. abscessus isolates were of the C28 sequevar and, thus, are sensitive to macrolides. Only the M. a. bolletii (Str 1) in our study contained the functional erm(41) gene (sequevar T28), which resulted in its resistance to clarithromycin in vitro after 14 days. Accordingly, the antibiotic therapy of this patient was the most complicated and lasted for 4 years.The crolides ,17. Thusequevar) . Our anaM. a. mycobacterioses. Treatment recommendations are based on case series, in vitro susceptibility testing, and the clinical experience of experts [M. a. infections. These can, however, frequently fail due to the development of resistance to macrolides by the mechanisms described above [M. a. massiliense, carrying the non-functional erm(41) gene [M. a. The patients were initially treated for a period of one month with intravenous amikacin and cefoxitin and oral clarithromycin. In the continuation stage, the combination of clarithromycin and doxycycline was used with the median duration of therapy being 24 months. As a result, the clinical recovery occurred in 83% patients, and conversion of the sputum to cultivation negativity occurred in 58%. However, among these, the conversion was significantly lower in patients infected with strains with demonstrated resistance to macrolides (17% versus 64%). On the other hand, when a group of patients was treated for subsp. massiliense infection, conversion of the sputum to cultivation negativity already occurred in the course of the first month of therapy, indicating the importance of the non-functional erm(41) gene in this subspecies [M. a. abscessus in those patients, who were infected by the sequevar C28 carrying the non-functional erm(41) gene [M. a.abscessus, where all 14 isolates possessed a non-functional erm(41) gene, and the conversion of sputum to cultivation negativity was achieved in 93% of patients.In general, mycobacterioses represent serious diseases, which are inherently difficult to treat . Current experts . Combinaed above . The usu41) gene ,31,32. T41) gene  describebspecies . This wa41) gene . Similar41) gene , in theiM. a. bolletii infection (Case 1), which was the only strain resistant to macrolides in our cohort. Case 1 was successfully managed by an extended therapy lasting 4 years, which was partly also due to a poor compliance of the patient. Patient compliance during the long-term antibiotic therapy is essential, as any misuse of antibiotics  can lead to the therapy failure. In this case, conversion of the sputum to cultivation negativity occurred after an improvement of patient’s compliance and the addition of clofazimine to the therapeutic regime.In three of our four treated patients, cultivation negativity was observed after the first 4 weeks of therapy. The repeated cultivation positivity was observed only in a patient with In the Case 2 patient, the result of therapy could not be evaluated, as she died after several months due to the GvHD refractory to therapy. The infections in the remaining two patients (Cases 4 and 5) were caused by strains susceptible to macrolides, and the introductory therapy led to both the clinical improvement and the conversion of the sputum to cultivation negativity after 4 weeks. These cases are, however, recent, and the continuation therapy is still ongoing .M. a. infections represent emerging infections in patients with CF or other chronic respiratory diseases [M. a. is usually identified only at the species level in routine laboratory practice [erm(41) gene and its sequencing was described as a powerful tool for the prediction of macrolide susceptibility and the quick determination of the initial therapeutic strategy [erm(41) gene correlate well with MIC values [M. a. strains were isolated from patients living in the East Bohemian region between years 2013 and 2021. In this study, the erm(41) gene analysis revealed a unique representation of M. a. strains, since, in two of them, an incomplete gene associated with macrolide susceptibility was found. Other two strains with complete but non-functional erm(41) genes were assigned as C28 sequevars. In only one strain, the inducible resistance was also proved (T28 sequevar). The correlation between erm(41) analysis and susceptibility determination to clarithromycin was excellent. Moreover, the erm(41) analysis significantly shortens the decision time for macrolide therapy, because inducible resistance is not detected until after 14 days. Although our group of patients is small, it represents all cases of M. a. infections identified in the Czech Republict to this date. Thus, it provides first record of this infection in the country and describes an obvious association between results of erm(41) gene analysis and clinical outcome.diseases . At prespractice ,34. In opractice ,36. Recestrategy . ResultsC values , which wClinical samples  were cultivated on solid media for 6 weeks at 37 °C under aerobic condition  and for 6 weeks at 37 °C in MGIT test tubes with modified Middlebrook broth in a semiautomatic system Bactec MGIT 960 .Mycobacterium species was performed by molecular typing. Nucleic acids were isolated directly from the suspension of suspect isolates or respiratory tract samples using a QIAamp DNA Mini Kit  according to the modified protocol for mycobacteria provided by the manufacturer.Identification of Mycobacterium using the PCR method . In case of positivity, further identification of Mycobacterium involved PCR amplification and sequencing by capillary electrophoresis of rpoB gene  as previously described [M. a. The M. a. subspecies were further typed by erm(41) gene amplification and sequencing according to Mase et al., 2011 [erm(41) gene with an incomplete product (product size 397 bp) identified M. a. massiliense. The full length erm(41) gene products (size 673 bp) indicated M. a. abscessus or M. a. bolletii, and further typing was based on the variability of nucleotides in select positions, as previously reported and shown in DNA of suspected isolates was identified as the species escribed . Final sl., 2011 . The ampM. a. was further confirmed by PCR method based on variable-number, tandem-repeat analysis, as previously described by Choi et al., 2011 [erm(41) and rrl genes  were userm(41) and rrl genes were performed in a volume of 25 µL containing buffer TaKaRa, 200 nM of dNTPs, 500 nM primers, and 1.0 U HS Taq polymerase TaKaRa. The amplification parameters were as follows: for erm(41) gene: 95 °C/10 min; followed by 95 °C/30 s, 60 °C/30 s, and 72 °C/30 s (35 cycles) finished by 72 °C/7 min and for rrl gene: 95 °C/10 min; followed by 94 °C/60 s, 55 °C/60 s, and 72 °C/60 s (35 cycles) finished by 72 °C/7 min. PCR products of both genes were visualized in agarose gel using an ethidium bromide. As positive control, the reference EQA sample of M. a. was used. Method was also validated on External Quality Assessment (EQA)—Quality Control for Molecular Diagnostics (QCMD) samples, where M. a. was correctly identified. Amplified DNA fragments of erm(41) and rrl genes were directly sequenced by capillary electrophoresis using primers identical to those used for PCR . The obtained nucleotide sequences were compared to reference sequences from GenBank  in an online tool, available at www.ebi.ac.uk/Tools/psa/emboss_needle. Sequence alignments were performed using DNAstar Lasergene software v. 17.3 , utilizing the Clustal W algorithm.PCR assays for Susceptibility of the isolates to antibiotics was examined by means of the E-test and the broth microdilution method. The results were interpreted according to the Clinical and Laboratory Standards Institute (CLSI). Selection of antibiotics included amikacin, cefoxitin, imipenem, clarithromycin, linezolid, doxycycline, tigecycline, ciprofloxacin, and moxifloxacin.MIC values of amikacin, cefoxitin, linezolid, doxycycline, ciprofloxacin, and moxifloxacin were established using the broth microdilution method with 96-well microtiter plates. Cation-adjusted Mueller–Hinton media  was used. Fresh colonies were harvested from the surface of solid media to saline. The bacterial suspension was homogenized on a vortex and diluted to a 0.5 McFarland standard density. The inoculum was prepared by a 1:100 dilution of 0.5 McFarland suspension in cation-adjusted Mueller–Hinton broth and added to plates.MIC values of imipenem, clarithromycin, and tigecycline were performed by E-test  on Mueller–Hinton agar plates . The bacterial suspension was prepared as above, and inocula were applied by swabbing according to Biehle et al. . The plaM. a. rank among the most difficult mycobacterioses to treat. The recommended treatment consists of a combination of antibiotics; however, the therapy can often fail due to the development of resistance to the key antibiotic, macrolide. The correct indication to the level of subspecies, testing of suitable antibiotics, and evaluation of sensitivity to clarithromycin after a prolonged incubation period are of principal importance for their management. Molecular detection of the erm(41) gene products and its sequencing can reliably and rapidly demonstrate the possibility of development of inducible resistance and thus influence the extent of introductory therapy and its duration. Our results show that the identification of the M. a. isolate to the level of subspecies is not necessarily sufficient. Only further analysis of the erm(41) gene in both isolates of M. a. abscessus allowed for the detection of the mutation, which resulted in the preserved sensitivity to macrolides. While this study shows only a limited number of cases, two out of five isolates (40%) carry a non-functional gene for the development of inducible macrolide resistance. This indicates that the share of such isolates with the preserved sensitivity to macrolides may be larger in such patients in our population than has been published so far. This should be considered when designing treatment strategies for the M. a. infections.Diseases caused by


Solidago canadensis L. and Solidago altissima L. are native to North America and have naturalized many other continents including Europa and Asia. Their species is an aggressive colonizer and forms thick monospecific stands. The evidence of the allelopathy for S. canadensis and S. altissima has accumulated in the literature since the late 20th century. The root exudates, extracts, essential oil and rhizosphere soil of S. canadensis suppressed the germination, growth and the arbuscular mycorrhizal colonization of several plants, including native plant species. Allelochemicals such as fatty acids, terpenes, flavonoids, polyphenols and their related compounds were identified in the extracts and essential oil of S. canadensis. The concentrations of total phenolics, total flavonoids and total saponins in the rhizosphere soil of S. canadensis obtained from the invasive ranges were greater than those from the native ranges. Allelochemicals such as terpenes, flavonoids, polyacetylene and phenols were also identified in the extracts, essential oil and the rhizosphere soil in S. altissima. Among the identified allelochemicals of S. altissima, the cis-dehydromatricaria ester may be involved in the allelopathy considering its growth inhibitory activity and its concentration in the rhizosphere soil. Therefore, the allelopathy of S. canadensis and S. altissima may support their invasiveness, naturalization and formation of thick monospecific stands. This is the first review article focusing on the allelopathy of both of S. canadensis and S. altissima. Solidago canadensis sensu lato (s.l.), belonging Asteraceae, is an erect rhizomatous perennial plant, native to North America. S. canadensis s.l. was introduced to Europe as an ornamental plant in the 17th to 18th centuries. The species spread from the gardens to the natural environments, and has extended its habitats in Central and Eastern Europe. It expanded its habitat at a rate of 741 km2 per year in Europe . A . A S. cativum L. . A flavo species ,77. Somenadensis .S. canadensis in allelopathy was also investigated. The production of the essential oil was greater in S. canadensis obtained from the heavily invaded stands than that from mildly invaded stands --2,8-decadiene-4,6-diynoate) and -10-angeloyloxy matricaria lactone were isolated from the stems, roots and/or rhizomes of S. altissima [Several polyacetylenes, such as ltissima ,104,111.ltissima ,112.S. altissima, cis-DME, dehydromatricaria lactone and the -10-angeloyloxy matricaria ester were reported to inhibit the coleoptile growth of Panicum crus-galli L. var. frumentaceum Trin. up to 77.4–93.5%, 81.7% and 80.0% that of the control, respectively, at the concentration of 1 ppm [Z,8Z)-10-Tigloyloxy matricaria ester also inhibited the growth of Lolium multiflorum Lam. [cis-DME among those polyacetylenes was the most studied.Among the polyacetylenes found in of 1 ppm ,111. Therum Lam. . Howevercis-DME caused the growth inhibition, which was of 1–20 ppm in laboratory conditions [cis-DME suppressed the germination of Asclepias syriaca L., Ambrosia artemisiifolia L. and Miscanthus sinensis Anderson; the germination and growth of Poa pratensis L. and Oryza sativa L.; and the growth of Lactuca sativa L. and Panicum crus-galli L. var. frumentaceum Trin. Their germination was inhibited up to 20–95% that of the control by 32–60 ppm cis-DME, and their growth was inhibited up to 10–56% that of the control by 48–100 ppm cis-DME  described above and the soil concentration (0–17.3 ppm) of cis-DME, which is the same in laboratory conditions, the cis-DME in the soil may have some ecological function.The concentration of ltissima  and of 0ltissima . The conltissima ,118,119.trans-DME was also in the soil under S. altissima, and its inhibitory activity was comparable of that of cis-DME [cis-DME was kept at 29 °C under 2000 lux for 2 days, 50% of cis-DME was isomerized into trans-DME, since the cis-form of the unsaturated ester is easily isomerized by light, pH or other conditions into the more stable trans-form [trans-DME was not found in the roots and rhizomes of S. altissima [cis-DME may be isomerized by light and other environmental factors into trans-DME after the exudation from the plants into the soil. When we consider the involvement of DME in the allelopathy of S. altissima, the concentration of both cis-DME and trans-DME in the rhizosphere soil should be counted. cis-DME . When thans-form . In addiltissima . TherefoS. altissima killed a pine wilt nematode (Bursaphelenchus lignicolous Mamiya and Kiyohara), and an active compound involved in it was determined as cis-DME. cis-DME at 10–11 ppm increased 50% of the molarity of the pine wilt nematode and a root-knot nematode (Meloidogyne incognita Kofild and White) [S. altissima rhizomes suppressed the hatching of fruit flies (Drosophila melanogaster Meigen), and an active compound was also determined as cis-DME [cis-DME may contribute to the establishment and naturalization of S. altissima in the introduced ranges.It was also reported that the aqueous extracts of the below-ground parts of d White) . The met cis-DME . The def cis-DME ,122,123.S. canadensis, the information on the allelopathic activity of S. altissima extracts, root exudates and residues is limited. However, polyacetylenes, monoterpene and sesquiterpenes, diterpenes, triterpenes and flavonoid glycosides were identified in its extracts and essential oil as allelopathic agents. Most potential compounds among them involved in the allelopathy may be cis-DME. cis-DME inhibited the germination and growth of several plant species at the concentration of 1–20 ppm [S. altissima and in its rhizosphere soil was of 250–400 ppm and 0–17.3 ppm, respectively [cis-DME may be released into the rhizosphere soil by root exudation, rainfall leachates and/or decomposition processes of plant residues. trans-DME was also formed by the isomerization of cis-DME in the soil, and its inhibitory activity was the same as cis-DME. Although there has been no information of the concentration of trans-DME in the rhizosphere soil of S. altissima, the concentration of only cis-DME in several soils was over the concentration which was able to cause the growth inhibition [cis-DME possesses nematicidal and insecticidal activities [S. canadensis [E-kolavenic acid also showed antifungal activity and insect antifeedant activity [trans-DME may be involved in allelopathy and/or defense function against natural enemies, such as fungal pathogens, parasites and herbivores, and may contribute to their establishment and naturalization into the introduced ranges. However, it is necessary to determine the concentration of trans-DME in the rhizosphere soil of S. altissima, as well as to investigate the allelopathy of S. altissima in greenhouse and field conditions. It is also worth investigating the effect of S. altissima on the arbuscular mycorrhizal colonization of the native plant species due to the fact that S. altissima and S. canadensis are very close species.Unlike 1–20 ppm ,111,113.ectively ,116. Thehibition ,116. In tivities ,124 Somenadensis . 13E-kolactivity ,110. TheS. canadensis and S. altissima are harmful invasive species naturalized in many countries, and form thick monospecific stands. Based on the literature, both species are allelopathic. The root exudate, residues, extracts, essential oil and rhizosphere soil of S. canadensis showed allelopathic activity on several plant species, including native plants, and suppressed the arbuscular mycorrhizal mutualism of the native plant species. Several allelochemicals were also identified in its extracts and essential oil. The concentrations of total phenolics, total flavones and total saponins in the rhizosphere soil of S. canadensis obtained from the invasive ranges were greater than those obtained from the native ranges. Therefore, the allelopathy of S. canadensis may contribute the interruption of the regeneration process of native plant species directly through the suppression of their germination and growth, and indirectly through the suppression of the mycorrhizal fungal mutualism of the native plant species. Several allelochemicals of S. altissima were also identified in its extracts and essential oil. Among them, cis-DME inhibited the germination and growth of several plant species at a concentration of 1–20 ppm, and its extractable concentration in the rhizosphere soil of S. altissima was of 0–17.3 ppm. cis-DME also exhibited nematicidal and insecticidal activities. trans-DME was formed by the isomerization of cis-DME in the soil, and its inhibitory activity was similar to that of cis-DME. Therefore, cis-DME, including trans-DME, may also cause the suppression of the regeneration process of the native plant species through the inhibition of their germination and growth. The allelopathy of S. canadensis and S. altissima may provide the species with competitive advantage against native plant species, and contribute to their invasiveness and naturalization.


Homoeusaovata . Paratypes. See, Type depository. The holotype is deposited at the Kyushu University Museum, Fukuoka, Japan.Etymology. The specific epithet ovata refers to the oval body shape of this new species.


Bivalve molluscs stand out for their ecological success and their key role in the functioning of aquatic ecosystems, while also constituting a very valuable commercial resource. Both ecological success and production of bivalves depend on their effective immune defence function, in which haemocytes play a central role acting as both the undertaker of the cellular immunity and supplier of the humoral immunity. Bivalves have different types of haemocytes, which perform different functions. Hence, identification of cell subpopulations and their functional characterisation in immune responses is essential to fully understand the immune system in bivalves. Nowadays, there is not a unified nomenclature that applies to all bivalves. Characterisation of bivalve haemocyte subpopulations is often combined with 1) other multiple parameter assays to determine differences between cell types in immune-related physiological activities, such as phagocytosis, oxidative stress and apoptosis; and 2) immune response to different stressors such as pathogens, temperature, acidification and pollution. This review summarises the major and most recent findings in classification and functional characterisation of the main haemocyte types of bivalve molluscs. Marine invertebrates constitute the largest group of macroscopic species in the sea . The phyHence, immune responses in bivalves and the processes that govern them, are important areas of active research. These immunological processes are centrally coordinated by a group of cells known as haemocytes which may act directly or in concert with humoral factors in the haemolymph to defend the animal against infection. Haemocytes constitute the cellular component of the haemolymph; they move through the circulatory system and migrate to other locations, such as the connective tissue and epithelia , 20. AmoRuditapes philippinarum granulocytes , enzymatic cytochemistry, physicochemical features and cell population separation, and biological activities and functions. Numerous authors have focused their efforts on developing a classification of the different blood cell types present in bivalves. The researchers Cheng (1981) and Hine (1999) published two of the most important reviews on morphofunctional aspects of haemocytes of the Phylum Mollusca . In the ulocytes  and up tseq data . One of seq data , 184; hoPerna perna  was not deemed totally correct, considering a haemocyte subpopulation as semi-granular cells  and granulocytes, there are some exceptions and not all haemocyte types occur in each bivalve species. In the clam rameters . Some aurameters , 39, 193rameters , thus a ar cells . Most ofar cells , 195. Alliformis . A partitype III , 78, andnulocyte .Another haemocyte type frequently found in bivalves is the blast-like cell Table 1,There is not a unified classification system for haemocytes in bivalve molluscs, thus far. On the contrary, the haemocyte types have been reported differentially between species. Even for the same species, different haemocyte classification has been proposed, as shown in M. galloprovincialis haemolymph, the occurrence of only one haemocyte type represented by two different ageing-related stages has been proposed; specifically, hyalinocytes in a proliferative stage which mature to become granulocytes . Various theories have been proposed to elucidate the lineage of haemocytes in bivalves. Cheng (1981) and Auffret (1988) propose two types of initial cell precursors capable of differentiating into granulocytes and hyalinocytes 8. Variou. Alternaulocytes , consistulocytes . In the an cells , 201, than cells . Unlike an cells , 74 leavecursors , 203. Th  and granulocytes, while acknowledging that these may be different life stages of the same cell type in some species. In spite that Cheng  suggesteIn the last two decades, the application of flow cytometry analysis and molecular characterisation of different immune-related molecules have greatly improved our knowledge of the functional characterisation of haemocytes, underlying both common and distinct features of the immune system in different bivalve species , 221. AlC. gigas  and to form pseudopods. Finally, granulocytes contained more mitochondria, which could provide energy in the phagocytosis process to result in higher phagocytosis ability . Howevercussatus . The graulocytes . In the ulocytes . Differeulocytes . Such obulocytes .A. farreri  seemed more diverse in protein content than the granulocytes in the oyster . edulis . In the inocytes . The difinocytes .C. ariakensis  and nitric oxide   and reactive nitrogen species (RNS), is induced after phagocytosis and it may be used to evaluate the immunocompetence in different haemocyte subsets . Oxidatiiakensis , C. gigaC. gigas , 251, C.ongensis , C. nipp nippona , C. virgirginica , 252, O.hilensis , Hyotiss kegaki , and S.lomerata ; in clamcussatus  and R. pppinarum ; in the  globosa ; in ark  granosa ; in mariincialis , M. unguiculatus , P. canaaliculus , P. viri viridis , and X.  securis ; and in rginalis . In L. m, a RNS) . The hig, a RNS) . Moreove, a RNS) , clams  , 256 and, a RNS) . These e, a RNS) , 259. Di, a RNS) ; in high, a RNS) .D. polymorpha, hyalinocytes showed the highest intracellular ROS production  than granulocytes -dependent and involve caspase-independent pathways  of the granulocytes and the erythrocytes type II increased linearly during the post-spawning period , thus these haemocyte types are potential players in biomineralisation processes  successfully identified numerous calcium-rich vesicles and crystals in granulocytes and assigned to this cell type the strongest ability of migration  and susceptible hosts . Oystersulocytes . In the  marinus . Similarcapacity . Oystersble ones , displayble ones . Remarkaulocytes ; similarexitiosa . In oystyte type  and highyte type . A positevalence .C. virginica haemocyte phagocytosis ability and granulocyte subpopulation  exposure . Exposurpulation . In the te count . Variatifew days . In the few days . Other rfew days , increasfew days , and chaC. gigas ; doublinppinarum ; decreasP. perna ; and anoP. perna  and variincialis . In vitrhilensis , while osponsive . Exposurlymorpha .C. virginica   and MPs increased granulocytes proportion  . In the increase . Nanoplalus spp. , 359, wis by NPs . Exposuroportion , while, oportion . In mussulocytes . Exposurinocytes . Ark clae to NPs –366 and e to NPs . SimilarHs alone . In the Hs alone . Long-tillutants .Considering the important effects of biotic and abiotic stressors on the bivalve haemolymph cells, extensive monitoring studies of the morphofunctional properties of the haemocytes of bivalves in the natural environment would be useful to state reliable criteria for diagnosing the physiological status of bivalves .Due to bivalve mollusc diversity and functional heterogeneity, haemocyte types vary from species to species and, nowadays, there is not a unified nomenclature that applies to all bivalves. Moreover, functions of each haemocyte type cannot be reliably extrapolated among all species. Haemocyte subpopulations own distinct properties that should be considered when characterising the overall immune related functions of bivalves. Granulocytes and hyalinocytes display differences in their metabolism and immune abilities, which implies that they play different physiological and immunological roles that should be deeply explored. Infections induce changes in the proportions of haemocyte types, which points out differential involvement of the haemocyte types in immune response. Abiotic stressors also alter the relative abundance of haemocyte types, which highlights functional differences. The ratio of cell types in the haemocyte community could be noted as indicator of immune function, being an important immune parameter to assess the bivalve health-status.NB: conceptualisation, methodology, literature review, and writing – original draft. FM: conceptualisation, literature review, writing – review and editing. AC: writing - review and editing. AV: conceptualisation, literature review, writing – review and editing, and supervision. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Geophagus altifrons  in Rio Jari, in the state of Amapá, in the eastern Amazon region. From the total of 31 fish examined, 90.3% were parasitized by one or more species, collecting a total number of 806 parasites. The parasites species identified were: Sciadicleithrum geophagi, Posthodiplostomum sp., Procamallanus  inopinatus, Raphidascaris (Sprentacaris) sp., Genarchella genarchella, Gorytocephalus spectabilis and Ergasilus xinguensis. Most of the parasites showed an aggregate dispersion pattern. Brillouin diversity index, uniformity and species richness of parasites were low. The component community of parasites was constituted by ectoparasites and endoparasites, but with a predominance of ectoparasites. The body size of hosts had a low effect on the parasites infracommunities. This first eco-epidemiological study for G. altifrons reports these parasites in a host, for the first time, with the exception of S. geophagi and E. xinguensis.The aim of this study was to investigate the ecology and diversity of community and infracommunities of metazoan parasites  Geophagus altifrons , an ornamental cichlid from the Amazon River basin in Brazil with sedentary habits, is a benthopelagic and omnivorous fish that feeds on large-bodied benthonic organisms, terrestrial and aquatic insects, crustaceans and fish, algae, detritus, macrophytes, fruits and seeds, depending on the seasonal period. This fish that inhabits lakes and rivers have a maximum length of 27 cm, and its first maturation occurs with 15-17 cm   , are theIn wild fish populations, communities and infracommunities of parasites are the result of repeated additions and losses of parasite species during evolutionary history. Developing over time, the ecological and biological characteristics of hosts and parasites determine host colonization, consequently influencing parasite community richness and diversity , b. The G. altifrons. The aim of this study was to investigate the ecology and diversity of community and infracommunities of metazoan parasites in G. altifrons of a tributary from the Amazon River system in northern Brazil.Host age, size, diet, habitat, behavior, distribution and geographical range have been recognized as some of the factors influencing richness and diversity of parasite communities in wild fish populations . TherefoG. altifrons (16.9 ± 2.7 cm and 108.6 ± 49.9 g) were collected in the Jari River basin, Jarilândia community, in the municipality of Vitória do Jari, Amapá state, Brazil  and with authorization from the Ethics Committee on the Use of Animals of Embrapa Amapá (Protocol No 014 - CEUA/CPAFAP).Fish were weighed (g) and measured in total length (cm) and then necropsied for parasitological analysis. The mouth, operculum, gills, viscera and gastrointestinal tract were examined in each fish to collection of ectoparasites and endoparasites, using a stereomicroscope. The collection, fixation, conservation, counting and preparation of the parasites for identification followed the recommendations of HB), evenness (E) and species richness  (rs) was used to investigate possible correlations of body length and weight of host with the richness of parasite species, Brillouin diversity index and parasite abundance  . Spearmabundance .G. altifrons, a total of 806 parasites were collected such as S. geophagi, Posthodiplostomum sp., Procamallanus  inopinatus , Raphidascaris (Sprentacaris) sp., Genarchella genarchella , Gorytocephalus spectabilis  and E. xinguensis  and species richness  with the length of hosts.There was no significant correlation of the Brillouin diversity index , abundance of Posthodiplostomum sp. , abundance of Raphidascaris (Sprentacaris) sp. , abundance of G. spectabilis  and abundance of G. genarchella  showed no correlation with host length. The abundance of E. xinguensis  presented a week positive correlation with the length of hosts, as well as the abundance of P. (S.) inopinatus . The abundance of G. spectabilis , abundance of S. geophagi , abundance of Posthodiplostomum sp.  and abundance of Raphidascaris (Sprentacaris) sp.  showed no correlation with the host weight. The abundance of E. xinguensis  and P. (S.) inopinatus  presented a week positive correlation with the host weight.The abundance of G. altifrons from Jari River, the community of metazoan parasites was composed by seven species, those being: Monogenea, two Digenea, two Nematoda, one Acanthocephala and one Crustacea; parasites with aggregate or random dispersion pattern. The community of metazoan parasites in Satanoperca jurupari  from Igarapé Fortaleza River was composed of eight species, those being: Monogenea, three Digenea, one Nematoda, one Acanthocephala and two Crustacea; all parasites with an aggregate dispersion  from the Igarapé Fortaleza River, the parasite community was composed of only three species, those being: one Monogenea, one Digenea and one Nematoda; parasites with aggregated dispersion   . In the nd world . The lifing bird . TherefoGenarchella genarchella occurred at low levels of infection in the intestine of G. altifrons from Jari River as reported for Hemibrycon surinamensis  from Igarapé Fortaleza River  inopinatus occurred at low levels of infection in G. altifrons. This nematode species, usually frequent in wild fish populations in Brazil, has fish as definitive hosts and in general has low levels of prevalence, intensity and abundance due to its complex life cycle (P. (S.) inopinatus increased with the weight and length of the hosts. It can be assumed that larger host fish are ingesting more infective stages of this nematode than smaller hosts. In addition, in G. altifrons of the Jari River, larvae of Raphidascaris (Sprentacaris) sp. are at infection levels lower than those reported for Geophagus proximus  found in the Tapajós River (PA) (Raphidascaris (Sprentacaris) have cladocerans as first intermediate hosts, smaller fish as secondary hosts and predatory fish as definitive hosts  . Speciesve hosts , 1998. GErgasilus xinguensis occurred at high levels of infestation in the gills of G. altifrons from the Jari River when compared to those reported for Geophagus argyrostictus  and G. altifrons from Rio Xingu, in the state of Pará (E. xinguensis for G. altifrons. Furthermore, abundance of E. xinguensis increased with the weight and length of hosts.Ergasilidae is one of the largest families of order Cyclopoida, and most species are found in freshwater fish. Just adult females have parasites in the gills, fins and nasal cavities of fish species . Ergasil of Pará . This isG. altifrons was composed by ecto- and endoparasite species with aggregate dispersion, but with a predominance of ectoparasites. This omnivorous fish occupies a lower position in the food web and is consumed by other larger fish species and fish-eating birds in the environment. The body size of hosts had a low influence on the parasites infracommunities. This first eco-epidemiological study of G. altifrons records, for the first time, these parasites for this host, except for S. geophagi and E. xinguensis.In conclusion, the component community of parasites in


M., female, born 1947, height 175 cm, weight 78 kg, BMI 25.4 was primarily admitted to the regional internal medicine department to rule out an acute coronary event. The patient suffered from attacks of acute shortness of breath, chest pain with repeated vomiting. She had a history of hypertension and hypoparathyroidism, treated pharmacologically; she had never undergone surgery. Cardiological examination ruled out a cardiological etiology. CT AG performed ruled out pulmonary embolism, but a bulky hiatal hernia with extensive gastrectasia was described , 2. Labo3In patients with hiatal hernias, patients with bulky paraesophageal, mixed hernias, and upside-down stomach are most often at risk for an acute course of the disease. Acute complications include gastric obstruction, acute bleeding, ischemia and gastric perforation. With gastric strangulation and ischaemia, patients are at risk of developing partial or complete gangrene of the stomach with subsequent perforation with the possibility of developing severe acute mediastinitis or peritonitis. In case of gastric obstruction with gastric contents, there is a risk of aspiration and development of pneumonia. The prevalence of acute conditions is reported in the literature to be 30.4 % 4In case of acute herniated hiatal hernia, it is always a serious condition with the possibility of developing complications. In case of acute symptomatology of the disease, surgical therapy is the only curative option. The procedure can be performed using a minimally invasive or conventional approach, with the nature and extent of the procedure modified according to the actual perioperative findings. This complication is not often described in recent literature and the results of treatment are not yet encouraging. Therefore, our solution can be considered as a recommended procedure and can successfully treat or even save the patient's life.Not commissioned, externally peer reviewed.There are no sources of funding for our research.Not required in our institution to publish Anonymous case reports.Written informed consent was obtained from the patient for publication of this case report and accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal on request.No registry.Ass. prof. Radek Vrba, MD, Ph.D. – design of the study, collection on the data, final approval of the version to be submitted.Ass. prof. JUDr. Dušan Klos, MD, Ph.D., MHA, LLM – design of the study, final approval of the version to be submitted.Daniela Kurfürstová, MD, Ph.D. – collection on the histopathological data, approval of the version to be submitted.Petr Špička, MD, Ph.D. – corresponding author, guarantor, revising the manuscript, final approval of the version to be submitted.Petr Špička, MD, Ph.D.Ass. prof. Radek Vrba, MD, Ph.D.The author declared no conflict of interest.


This is a correction of: Mladinić A, Puljak L, Koporc Z. Post-GDPR survey of data protection officers in research and non-research institutions in Croatia: a cross-sectional study. Biochem Med (Zagreb). 2021;31:030703. 10.11613/BM.2021.030703.Since the publication of the article, the authors have noticed that one sentence was not correct, as the legislation has changed before the manuscript was published. Thus, in order to provide accurate information to the readers, the sentence: “According to the Personal Data Protection Act of Croatia, The Personal Data Protection Agency shall keep a Register of Personal Data Protection Officials.” should be disregarded. The authors apologize for any inconvenience caused to the readers.


Escherichia coli (E. coli) is a common pathogen that causes diarrhea in humans and animals. In particular, E. coli can easily form biofilm on the surface of living or non-living carriers, which can lead to the cross-contamination of food. This review mainly summarizes the formation process of E. coli biofilm, the prevalence of biofilm in the food industry, and inhibition methods of E. coli biofilm, including chemical and physical methods, and inhibition by bioactive extracts from plants and animals. This review aims to provide a basis for the prevention and control of E. coli biofilm in the food industry. Escherichia coli (E. coli), a member of Enterobacteriaceae, was discovered by Escherich in 1885. It is 0.5 × 1-3 μm in size with peritrichous flagellae [E. coli can also ferment a variety of sugars to produce acid and gas. Certain special serotypes of E. coli were recognized as pathogenic and capable of causing human- and animal-infections by the mid of 20th century. In 1982, E. coli was first defined as an intestinal pathogen and an important source of food-borne diseases. These bacteria can cause serious diseases, such as hemorrhagic proctitis, hemolytic uremic syndrome, and acute renal failure. Additionally, E. coli can easily form biofilms, which also affects human health [E. coli’s pollution, the incidents of food poisoning by the contamination of E. coli are still common, and have caused serious damage to human health, as well as huge losses to the food industry and economy [E. coli outbreak in 13 states in North America, which caused multiple infections [E. coli (EHEC) disease. Consumers ate contaminated bean sprouts, which caused diarrhea, hemolytic uremic syndrome and even death [E. coli O157 food poisoning occurred in dozens of middle schools and kindergartens in Okayama, Hiroshima, and other areas of Japan, and the number of infections reached nearly 10,000 [At present, despite plenty of research for strengthening the prevention and control of  economy . In 2018fections . In 2011en death ,7. In 19E. coli can greatly increase its resistance to environmental stress and often results in sterilization failures. Biofilm is composed of microbial colonies and self-produced extracellular polymeric substances (EPS), where the microorganisms of the same or different species are spontaneously wrapped in their self-secreted EPS matrix, which provides resistance to the external environment, and attachment to the surface of living or non-living carriers [E. coli biofilm, which could provide novel insights into the control of E. coli biofilm in the food industry.Notably, the biofilm-formation capability of carriers ,10,11. TThe formation of microbial biofilm is a universal phenomenon. As long as the conditions permit, microorganisms often form biofilm in the environment . The forE. coli biofilm is generally divided into five stages, including reversible adhesion, irreversible adhesion, colony formation, biofilm maturation, and dispersion, as shown in Since the observation of microbes and biofilms by Antony van Leeuwenhoek in dental plaques using a microscope in 1676, studies about the mechanisms of biofilm formation have evolved rapidly . To dateThe free microorganisms receive environmental signals (such as nutrients), and use their extracellular organelles  and outer membrane proteins in order to adhere to the carrier surface.Microorganisms secrete EPS, which enhances the adhesion of microorganisms with the carrier surface. EPS is composed of nucleic acids, proteins, lipids, lipopolysaccharides, and other substances, and forms a protective barrier for microorganisms by restricting and preventing the antibacterial agents from reaching the biological membrane of target organisms ,15. NotaE. coli into the biofilm lifestyle by regulating the synthesis of biofilm matrix compounds [The microorganisms, adhering to the carrier surface, grow and multiply to form colonies. Then, the continuous proliferation of microorganisms makes their biological growth space crowded with a lack of nutrients and accumulation of toxic substances. At this stage, the quorum-sensing (QS) signals, a cell-to-cell communication mechanism in bacteria, could induce the ompounds . During ompounds .E. coli microcolonies continue to accumulate, which further increases the thickness of the film and results in the formation of mushroom-like or columnar subunits. Then, the biofilm grows in a three-dimensional manner and eventually forms a viable three-dimensional structure [tructure .The dispersion of biofilm can be divided into active dispersion and passive dispersion. The active dispersion refers to a decrease in the level of cyclic diguanylate (c-di-GMP) in cells, which results in the production of enzymes that degrade the matrix of biofilm and promote its dispersion. The passive dispersion depends on the external factors, such as physical triggers  and enzymatic degradation (extracellular hydrolase) . The disE. coli biofilm is a strong survival support for E. coli in unfavorable living environmental conditions, protects the growth and reproduction of E. coli to the greatest extent, and provides favorable conditions for E. coli to wreak havoc on and pollute food, eventually causing infection and affecting human health.In conclusion, the formation of E. coli is an indicator of fecal pollution, and a serious threat to the life and health of consumers. Once the content of E. coli in a food item exceeds the standard, it is withdrawn from the shelves, resulting in huge economic losses.Hygienically, E. coli may pose a greater threat to the processing and production of food. Studies show that once the microbial biofilm is formed, the resistance of bacteria to disinfectants is increased by 500 times, suggesting that, in the presence of biofilms, the germicidal time and concentration of disinfectants must be increased to 10–100 times to kill bacteria [Moreover, the biofilm-making ability of bacteria ,22. Anotbacteria .E. coli biofilm exists in nearly all aspects of the processing and production of all kinds of food , and may cause pipeline biological pollution and equipment damage , thereby contaminating food items [At present, a couple of studies have shown that the od items ,25,26,27E. coli, as it grows in a natural environment. This may introduce E. coli into the food industry. Studies have reported the isolation of E. coli from peppers, tomatoes, cantaloupes, and other crops in Nuevo Leon and Coavira, Mexico. A total of 341 strains of E. coli have been isolated, of which 76% of strains have the ability to form biofilms. Among them, 34% of E. coli strains form strong biofilms [E. coli to cause human and animal diseases, a great importance has been given to the detection of E. coli in poultry products. At present, Brazil is the largest exporter of chicken. Among the 88 strains of E. coli isolated from the chicken meat of four retail chicken companies in Brazil, 31 strains could form a strong biofilm, while only four strains could not form biofilm [E. coli can easily form biofilms in processing equipment, such as stainless-steel metal products [E. coli from fresh-cut processing equipment. The co-culturing of E. coli with the isolated Burkholderia caryophylli and Ralstonia insidiosa increases biofilm biomass by 180% and 63%, respectively, posing a great challenge to eliminating the foodborne pathogens in fresh-cut processing equipment [Food ingredients can also be easily contaminated by biofilms , which p biofilm . In addiproducts ,30, polyproducts , due to quipment .E. coli can form biofilm on the surface of food items, pipes, and processing equipment, which greatly increases its resistance to environmental conditions and reduces the effectiveness of disinfectants [E. coli in food items and harms the health of consumers. Therefore, it is urgent to take effective measures to reduce or control the formation of E. coli  biofilm during processing, as shown in fectants . The bioE. coli biofilm to a certain extent, but cannot completely inactivate it [E. coli biofilm, the inhibitory effects of NEOW and CD on E. coli biofilm biomass were significantly higher than that of SH and exhibited a certain degradation effect on E coli biofilm. Among them, NEOW was the most stable at a low temperature (5 °C) with the lowest chlorine loss rate and a wider bacteriostatic temperature range [E. coli biofilm suggested that the HPQM exhibited comparatively better inhibitory properties and bacteriostatic effects on LLDPE with low roughness than ZEOMIC [2 and SH were found to show the highest anti-biofilm activity against E. coli on stainless steel, followed by glass, plastic, and wood [E. coli on the surface of Hami melon, but a 2000-μg/mL concentration of SH and arginine LAE had no obvious effects on the inhibition of E. coli biofilm, and the biofilm showed high resistance to disinfectants [E. coli biofilm and the mixed biofilm of E. coli and Staphylococcus aureus. Its inhibitory effect is mainly through the rise of reactive oxygen species, leading to bacterial oxidative stress and damaging the respiratory system [At present, the disinfectants, commonly used in food processing and slaughterhouses, include chlorine disinfectants, quaternary ammonium chloride (QAC), and lactic acid, which inhibit the formation of ivate it . When stre range . The comn ZEOMIC . At the and wood . This infectants . Therefoy system .E. coli biofilm in food processing and production, physical methods  are often used in addition to chemical methods to reduce the occurrence of food’s cross-contamination. High-temperature sterilization is the most common physical method to control microorganisms. Saturated steam (SS) and superheated steam (SHS) can inactivate polyvinyl chloride (PVC) in food processing facilities and inhibit the formation of E. coli biofilm on the surface of stainless steel. SHS showed higher lethality and better inactivation of the cells in E. coli biofilm [E. coli biofilm. When the amplitude of vibration on the surface is greater than 21nm, the greater the amplitude, the less the number of viable bacteria. It can provide a reference for some precise and difficult methods to clean mechanical surfaces in food or medicine [E. coli biofilm and a reduction in E. coli’s abundance under the biofilm to 5log10. At the same time, the low-temperature plasma sterilization, in combination with water spray, can not only inhibit bacteria, but also improve the efficiency of antifouling [E. coli biofilm and Listeria biofilm, while this inhibitory effect slightly decreased as biofilm growth age increased. Afterwards, a mixed culture of the two bacteria showed that the production of EPS in E. coli biofilm may contribute to the increases in the resistance of cold atmospheric plasma, which provides a new target to inactivate bacteria on the complex solid surface [the Shiga toxigenic E. coli (STEC) in the biofilm on stainless steel would not transfer to lettuce. However, when the temperature is 25 °C, STEC will transfer from the biofilm to stainless steel. It is suggested that keeping a low temperature in a food processing environment is helpful to control the transfer of E. coli in biofilm, resulting in microbial cross-contamination [For the inhibition of  biofilm . To prevmedicine . In addimedicine . The lowifouling . Similar surface . In addimination .E. coli biofilm. The combination of lactic acid (LA) with water vapors can effectively reduce the abundance of E. coli on the surface of PVC and stainless steel, and exhibits lethal effects on the microorganisms in biofilm [E. coli biofilm [In the actual food processing and production, food industries often use the combination of chemical and physical sterilization methods to control microorganisms, and the synergistic effect between them maximizes the removal or inhibition of  biofilm . The com biofilm .E. coli biofilm have been widely reported.In recent years, due to the decrease in the sensitivity of microorganisms in biofilm to conventional disinfectants, the germicidal efficacy of conventional disinfectants cannot reach an ideal state, which increases the risk of food microbial safety. Therefore, there is an urgent need to seek new active germicidal substances and improve their efficiency. Biological extracts are natural compounds, which avoid the problem of chemical disinfectant residues and have attracted wide attention because of their higher level of safety. Therefore, the inhibitory effects of biological extracts on E. coli biofilm have attracted the attention of researchers. The main component of scallop shell is calcium carbonate (CaCO3), which is easily decomposed into calcium oxide (CaO) by heating, which then exhibits broad-spectrum antibacterial properties [E. coli biofilm on the surface of stainless steel in whey powder solution and meat processing plant washing water by 4 and 6 log CFU/cm2 and 3 and 5 log CFU/cm2, respectively. This inhibitory effect was positively correlated with the concentration of scallop shell powder, suggesting that it can be a good candidate for application in the food industry [With the attitude of safety, environmental protection, economy, and turning waste into treasure, animal extracts such as the scavenging effects of scallop shells on operties ,62. Studindustry .10 CFU, thereby reducing the risk of food cross infection [E. coli biofilm, such as bacteriophage FP43, Daica, and135 [In addition, the concept of using bacteriophages, the natural predators of bacteria, as a novel strategy to prevent and eliminate biofilms has gradually attracted the attention of researchers. The treatment with phage AZO145A significantly inhibited the biofilm formation of STEC on the surface of stainless-steel plate, and significantly inhibited their migration at 24 °C in the biofilm formed in beef, which was reduced to 3.1 log, and135 ,46. In t, and135 .E. coli have been a hot topic. The plant extracts can be directly used in food processing equipment or food items to inhibit the E. coli biofilm and prolong the shelf-life of food items. Phenols are widely studied for their inhibitory effects on E. coli biofilm. Four components were obtained after extracting the active components from burdock leaves with ethanol and diluted with water to form different concentrations of ethanol . It was found that the active components obtained by different ethanol elution could significantly inhibit the E. coli biofilm, where the inhibitory effects were correlated with the phenol contents in the burdock leaves. The 70% ethanol elution could seriously damage the structure of E. coli biofilm, which no longer showed the state of a multi-layer growth [E. coli biofilm [E. coli biofilm, which were proportional to the time, and could completely remove the biofilm after 15 min of exposure [E. coli biofilm without affecting the growth of bacteria [E. coli biofilm. The combination of clove oil and liposome, making solid liposomes (SLPs), can avoid the disadvantage of volatilization under light and at a high temperature, and improve stability and the utilization rate. The SLPs treatment dispersed the E. coli biofilm from the whole to a loose state, which seriously damaged its structure, decreased the biomass under membrane, and saw some cells collapse. In addition, the application of SLPs to the surface of cucumber and lettuce also showed a good inhibitory effect on biofilm and prolonged the shelf life of vegetables [E. coli biofilms, with an inhibition rate of 50% at 6.25 g/mL or more [The inhibitory effects of plant extracts on the biofilm of r growth . The com biofilm . The lowexposure . The chabacteria . In addigetables . The ant or more .E. coli biofilm mainly depend on the inhibition of cell metabolic activities; gene expression; reduction in extracellular polysaccharides, bacterial motility, and cell hydrophobicity; and directly preventing the initial adhesion stage of biofilm formation. The treatment of E. coli with SLPs suggested a negative correlation of the SLPs concentration of clove oil with the bacterial metabolic activities, extracellular polysaccharides, and protein contents, thereby weakening the protective barrier and adhesion of bacteria [Coptis chinensis could also inhibit the metabolic activities of E. coli biofilm [E. coli to the surface; and inhibited the formation of biofilm [E. coli biofilm with triterpenoids (oleanolic acid and ursolic acid) suggested that oleanolic acid analogues could change the expression of genes involved in the regulation of type IV pili [E. coli by inhibiting the formation of biofilm, and ETH, MEC, TEB, and PYR could significantly reduce the surface hydrophobicity of E. coli cells and hinder the adhesion stage during the formation of E. coli biofilm [E. coli, vitamin C can down-regulate the signal transduction genes and regulatory genes of biofilm by more than 27 times [E. coli and Listeria on the surface of polypropylene, it was also found that the inhibitory effect of cinnamon, marjoram, and thyme could be achieved by penetrating the biofilm and cell membrane, further changing the fluidity and permeability of the membrane, condensing the protons of the cytoplasm to form the weak mitochondria [The inhibitory effects of plant extracts on bacteria . The ext biofilm . Coumari biofilm . Studyin IV pili . CAT, TE biofilm . In the 27 times . In the chondria .E. coli biofilm has become a hidden danger of microbial contamination in the food industry. The formation of E. coli biofilm is a complex process, which is affected by a variety of regulatory networks. At present, there is an urgent need for studies on the regulatory mechanism of biofilm maturation, as well as the prevention methods of biofilm formation. Meanwhile, studying the mechanisms of the inhibition of E. coli biofilm is of great significance for the development of efficient and safe germicide to reduce E. coli biofilm in the food industry.The formation of microbial biofilm is a kind of self-protection behavior of microorganisms in unfavorable environmental conditions, which helps them in avoiding environmental stresses. Therefore, the


Dermestes (s.str.) haemorrhoidalis—a species considered relatively rare in Poland, whose presence on human corpses was first observed in 2020 in Wroclaw.Dermestidae are beetles of economic importance but also of significance for forensic entomology. In the latter case, Dermestidae are classified as colonisers of cadavers in late stages of decomposition when the remains are in a dry phase (mummified human corpses) or are partially skeletonised. Therefore, they are classified as secondary necrophages. This article describes three cases of mummified human corpses in indoor conditions that were colonised by representatives of the Dermestidae. All three cases are linked by the presence of Dermestes Linnaeus, 1758, are often identified on corpses in late stages of decomposition. They usually feed on remains devoid of organs and soft tissues or when the corpses undergo mummification. In Europe, eight species from two subgenera Dermestes and Dermestinus Zhantiev, 1967, have so far been identified on human corpses. Despite the relatively frequent presence of Dermestes sp. in experimental studies conducted in Poland, no reports concerning Dermestes directly collected from human corpses have been published to date. This article again describes observations of Dermestidae collected from human corpses found in indoor conditions in Wrocław, the capital of the Dolnośląskie Voivodeship. For the second time, there is evidence of the presence of Dermestes (s.str.) haemorrhoidalis on human corpses—a species considered to be relatively rare, as evidenced by faunistic data published from Poland, as well as the results of ongoing experiments of forensic interest.Beetles of the family Dermestidae, especially of the genus  Dermestes Linnaeus, 1758 (Coleoptera: Dermestidae) are necrophagous insects found on corpses, including human corpses [Dermestes includes approximately 90 species, including fossils ones [Dermestes (s.str.) haemorrhoidalis Küster, 1852. This species, described from southern France, mainly inhabits southern Europe while data on its distribution are fragmentary, especially in central Europe. Moreover, this species has been reported in Africa , South America , and Asia  [D. (s.str.) haemorrhoidalis was first found in Warsaw, where it was brought in the 1960s and where it probably acclimatised [Beetles of the genus  corpses ,3,4. The corpses . Informa corpses  and Kadeils ones . An examVietnam) ,11,12. IVietnam) . StudiesVietnam)  and JakoVietnam) . In Polaimatised . The disimatised  and dataDermestes (s.str.) haemorrhoidalis was reported.This article describes below three cases of mummified human corpses from south-western Poland from the city of Wrocław, in which the presence of Dermestes sp. found in Poland on animal or human corpses both under experimental conditions and in available case studies.In this article, the species of associated insects are only briefly described. This article includes comparisons of published data on Insects were stored in plastic vials for morphological examination. The studied material was collected directly from the corpse in an autopsy room (cases #1–3) or at the place where the corpse was found . Species identification was conducted using a Nikon SMZ800  binocular microscope. All materials are stored at the Centre for Forensic Biology and Entomology, University of Wrocław.Both dead and live larvae/adults were preserved in 75% ethanol. Larval moults, after drying, were stored in plastic boxes. For final species identification, male genitalia were separated and dissected and then preserved in warm 10% KOH and washed in distilled water.Species identification was conducted using a Nikon SMZ800  binocular microscope. The identification of Dermestidae was made by the analysis of specific species characteristics of both adults and larvae, including the comparison of male genitalia ,6,17. Ot® 4500  digital camera. Image stacks were processed using Combine ZP® software .Images were taken under a Nikon SMZ800  binocular microscope with a Nikon CoolpixOn 10 August 2020, the corpse of an 83-year-old single man was discovered in a flat on the first floor of a multifamily two-story house in an old (before World War II) building in the centre of a large city . The corD. (Dermestinus) frischii Kugelann, 1792 (Coleoptera: Dermestidae) (D. (s.str.) haemorrhoidalis (Coleoptera: Dermestidae). Adults of three Dermestidae species of the genus Dermestes (Coleoptera: Dermestidae) were collected from the flat of the deceased and preserved. They included D. (D.) frischii , D. (s.str.) lardarius Linnaeus, 1758 (Coleoptera: Dermestidae) , and D. (s.str.) haemorrhoidalis . Attagenus smirnovi Zhantiev, 1973 (Coleoptera: Dermestidae) (multiple live larvae) and Anthrenus verbasci  (Coleoptera: Dermestidae) (one live larva) were also collected from the flat and preserved. Puparia and dead adults of flies Calliphora vicina Robineau-Desvoidy, 1830 , Lucilia sericata  , and dead adults of Sarcophaga Meigen, 1826 (Diptera: Sarcophagidae) were present in large numbers on windowsills, furniture, and the floor .In an autopsy room, mostly dead adults, larvae, and larval moults of two Dermestidae species were collected from the corpse and preserved, i.e., estidae)  and D.  haemorrhoidalis  were collected and preserved. In this case, the forensic entomologist was not present at the site where the body was found.In terms of insects found on the corpse, adults and larvae of On 26 February 2021, the corpse of a 75-year-old single woman was discovered in a flat on the fifth floor of a block of flats dated around 2000. The deceased lay on her back on the kitchen floor; the flat was in disarray. The woman did not maintain contact with her family and neighbours. In March 2019, there was an emergency room intervention for her due to cardiac problems. A neighbour living opposite to the flat where the body was found last saw the deceased about two years earlier; he used to see her very rarely. Autopsy findings included advanced late post-mortem changes in the form of natural mummification with soft tissue loss involving the head, neck, shoulders, and upper thoracic region. The residual preservation of internal organs included a transformed brain with completely blurred structure and a homogeneous conglomerate of highly transformed trunk organs that were impossible to identify. Insects and evidence of their development and feeding (Coleoptera and Diptera) were found on the corpse . The weiNecrobia rufipes  (Coleoptera: Cleridae) (eighty-five adults and twenty larvae) and D. (s.str.) haemorrhoidalis  were collected from the corpse in an autopsy room and preserved. In addition to beetles, developmental forms of the following Diptera were collected: Piophila casei  (Diptera: Piophilidae) , puparia of Phoridae , and Chrysomya albiceps   (numerous puparia and three dead adults). In this case, the forensic entomologist was not present at the site where the body was found.Dermestes were identified on human corpses: D. (Dermestinus) frischii; D. (D.) maculatus DeGeer, 1774; D. (D.) undulatus Brahm, 1790; D. (s.str.) ater DeGeer, 1774; D. (s.str.) bicolor bicolor Fabricius, 1781; D. (s.str.) haemorrhoidalis; D. (s.str.) lardarius; and D. (s.str.) peruvianus Laporte de Castelnau, 1840 [D. (s.str.) haemorrhoidalis was mainly found on dry or skeletonised corpses decomposing in indoor conditions [D. (s.str.) haemorrhoidalis was the only representative of the genus Dermestes, represented by larval moults, live larvae, dead and live adults, and frass. Similarly, in the three cases presented in this paper, D. (s.str.) haemorrhoidalis was the species found on the corpses and was even the dominant species . All of the above-mentioned cases of corpse mummification took place in the urban agglomeration and inside living quarters.In Europe, eight species of the genus au, 1840 . The aboA. smirnovi, A. verbasci, and C. vicina, the following species were also collected from the flat and corpse and preserved: D. (D.) frischii, D. (s.str.) lardarius, L. sericata, and Sarcophaga sp. As the study by Lutz et al. proved [L. sericata and C. vicina and beetles such as D. (s.str.) lardarius were the most abundant species on human corpses analysed in Germany. In case #2, the species composition was dominated by D. (s.str.) haemorrhoidalis. This situation can be explained by specific circumstances, e.g., the hot and dry environment, rapid mummification of the body, and the absence of other necrophagous insects. In such conditions, Dermestes sp. can colonize a body in huge numbers [D. (s.str.) haemorrhoidalis still included species such as N. rufipes, P. casei, Phoridae , and Ch. albiceps. In the light of published data from Europe, the above-mentioned assemblage is fairly typical of indoor decomposition, excluding Piophilidae, which are considered rather typical of outdoor decomposition [Chrysomya albiceps, although usually found in environments such as meadows and forests [Necrobia rufipes can also be found on both animal [N. rufipes in relation to Dermestidae are also consistent with the results of a study by Charabidze et al. [Necrobia spp., although it was observed especially for outdoor varieties.Case #1, which was described in this paper, had a composition of insect species found on corpses that was strongly similar to the assemblage reported from Wrocław by Kadej et al. . The dif. proved , Diptera numbers ,22. In cposition , as confposition ,25,26,27 forests ,24,28,29 forests ,28,30. S forests ,31,32. Nh animal ,26,27 anh animal ,33. ObseDermestes (three species) on the corpse. In the other two cases (cases #2–3), only D. (s.str.) haemorrhoidalis was found on the corpses. These observations are consistent with the results from northwestern Europe, where the highest percentage of findings was for only one species of Dermestes sp. on corpses, regardless of whether decomposition occurred in indoor or outdoor conditions [In the described cases, only in case #1 there was more than one representative of Sus scrofa domestica Linnaeus, 1758) carcasses in outdoor conditions detected the following representatives of the genus Dermestes: D. (D.) frischii [D. (D.) murinus murinus Linnaeus, 1758 [D. (D.) laniarius laniarius Illiger, 1802 [D. (s.str.) lardarius, D. (D.) frischii, and D. (D.) undulatus were also found on a male corpse in outdoor conditions [D. (s.str.) lardarius on a corpse pulled from a chimney [D. (s.str.) haemorrhoidalis prefers corpses in indoor conditions, such as rooms in residential buildings. This is also confirmed by previously published faunistic data from Poland [D. (s.str.) bicolor bicolor. In one of the three cases described in this study (case #1), this species was in turn accompanied by D. (D.) frischii and D. (s.str.) lardarius. The latter observation partly coincides with the result obtained by Voigt [D. (s.str.) lardarius was present on a corpse together with D. (s.str.) haemorrhoidalis.Data from Poland  from expfrischii ,26,27,34us, 1758 ,36, and nditions   peruvianus was the dominant species on corpses in indoor conditions [D. (s.str.) haemorrhoidalis [In contrast to results from northwestern Europe where nditions , observahoidalis . ConfirmD. (s.str.) maculatus [In all three cases described in this study (cases #1–3), there was only the damage to the dehydrated skin typical of Dermestidae (cf. ). Only iaculatus ), no sucaculatus . The lacD. (D.) frischii, in the empty shells of pupae of flies of the family Calliphoridae (D. (s.str.) haemorrhoidalis, also collected from a mummified corpse found in one of the flats in Wrocław [D. (D.) frischii exhibits predatory behaviour in relation to larvae of Diptera, as experimentally proven for D. (s.str.) ater in relation to larvae of Musca domestica Linnaeus, 1758 (Diptera: Muscidae) [Dermestes sp. found on human corpses.An interesting observation was made in case #1, namely, the confirmation of pupation, this time by phoridae . A simil Wrocław . The eviuscidae) . These oD. (s.str.) haemorrhoidalis in all so far published descriptions of mummified human corpses in indoor conditions in Wrocław ([D. (s.str.) haemorrhoidalis on corpses prove that this species, contrary to what Mroczkowski wrote in 1975 [Columba livia f. urbana Gmelin, 1789 (Aves: Columbiformes: Columbidae), whose nests are very frequently built in residential buildings in Wrocław , may be one of such sources. The possibility of the colonisation of bird nests by this species was mentioned by Hockin [D. (s.str.) haemorrhoidalis might breed in pigeon nests in London. From bird nesting sites, D. haemorrhoidalis may be attracted to corpses not only by the smell of decaying remains, but also by a male-produced aggregation pheromone [A separate commentary is required regarding the interpretation of the result of the presence of Wrocław  haemorrhoidalis has so far been found sporadically in Poland, and knowledge of its distribution requires further in-depth faunistic studies. The results of autopsies of mummified human corpses, when confronted with data from field experiments, support the thesis that this is a species which mainly prefers decomposing corpses in indoor conditions. The cases presented by us indicate that D. (s.str.) haemorrhoidalis in this part of Poland may be a common or even the only larder beetle on corpses in the late stage of decomposition. This indicates the potential of this species in a forensic context. For this reason, further studies regarding the biology of this species found on human corpses would also be advisable, including studies in terms of its interactions with other species. According to recent scientific reports, it is also worth investigating the potential use of bones as material for the creation of pupal chambers by this species. The results described above also reveal the need to collect and preserve entomological material not only from human corpses in an autopsy room, but also directly at the site where the corpse was found. A visit by an entomologist to the site where a corpse was found not only allows more entomological evidence to be preserved, but also gives the opportunity to gather more information on species of forensic importance, their behaviour, and their use of the space around the corpse. For this reason, it should be good practice to allow a forensic entomologist or other competent person to participate in the examination of the body.In the future, based on cooperation between institutions such as the police and the prosecutor’s office, the intention is to focus on Dermestidae that colonise human corpses in order to carefully analyse their communities in both outdoor and indoor conditions in Poland. This is particularly necessary because despite the relatively frequent occurrence of Dermestidae on human corpses, few studies analyse the communities of Dermestidae in detail.


A 57 year-old man was referred for renal transplantation due to diabetic nephropathy. He was transferred from the city of Macapá, in the Brazilian Amazonian region. He reported a history of multiple chronic keloid-like lesions since he was three years-old. Along his life these lesions progressed to involve his face, trunk, and members. Lesions varied in size, some were confluent, nodular and ulcerated. At the age of 16 he was diagnosed with lobomycosis , after a skin biopsy was performed. He received courses of dapsone, itraconazole and clofazimine with no success. His social life was very much impacted by the disease. At arrival in our medical center, we had extensive and itchy, disseminated keloids . He was