Antimicrobial Resistance in Enterobacterales Bacilli Isolated from Bloodstream Infection in Surgical Patients of Polish Hospitals.

BACKGROUND AND AIMS: Bloodstream infections (BSIs) are one of the most frequently observed hospital-acquired infections (HAIs). We sought to describe the epidemiology and drug resistance secondary Enterobacterales BSIs in surgical patients and check for any correlation with the type of hospital ward. MATERIALS AND METHODS: This multicenter (13 hospitals in southern Poland) laboratory-based retrospective study evaluated adults diagnosed with BSI secondary to surgical site infection (SSI) hospitalized in 2015-2018; 121 Enterobacterales strains were collected. The drug resistance was tested according to the EUCAST recommendations. Tests confirming the presence of extended-spectrum ß-lactamases (ESBLs) and bla resistance genes were carried out. The occurrence of possible clonal epidemics among K. pneumoniae strains was examined. RESULTS: The prevalence of Enterobacterales in secondary BSI was 12.1%; the most common strains were E. coli (n = 74, 61.2%) and Klebsiella spp. (n = 33, 27.2%). High resistance involved ampicillin and ampicillin/sulbactam (92, 8-100%), fluoroquinolones (48-73%), and most cephalosporins (29-50%). Carbapenems were the antimicrobials with the susceptibility at 98%. The prevalence of ESBL strains was 37.2% (n = 45). All the ESBL strains had bla CTX-M gene, 26.7% had the bla SHV gene, and 24.4% had bla TEM gene. The diversity of Klebsiella strains was relatively high. Only 4 strains belonged to one clone. CONCLUSIONS: What is particularly worrying is the high prevalence of Enterobacterales in BSI, as well as the high resistance to antimicrobial agents often used in the empirical therapy. To improve the effectiveness of empirical treatment in surgical departments, we need to know the epidemiology of both surgical site infection and BSI, secondary to SSI. We were surprised to note high heterogeneity among K. pneumoniae strains, which was different from our previous experience.

Concurrent Infection of a Young Tourist by Hookworm and Strongyloides Stercoralis During Low Budget Travel in Southeast Asia.

Strongyloidiasis and hookworm infections are neglected helminth diseases widespread in tropical and subtropical areas. In humans, particularly in immunocompromised patients infections potentially may lead to the life-threatening clinical conditions involving the respiratory as well as gastrointestinal systems. The increased number of tourists travelling to tropical regions is associated with more frequent infection with parasites such as Strongyloides and hookworm. The infection takes place when filariform larvae penetrate the skin exposed to soil, than migrate through the lungs and finally reach the intestine. Travelers are often not aware of how they could get infected. Physicians may suspect strongyloidiasis and hookworm infections in tourists with diarrhea returning from endemic areas, especially when an elevated eosinophilia is observed. In the literature there are many reports about the presence of parasites in indigenous communities, but very few are available regarding travelers. This paper describes a dual infection with hookworm and Strongyloides stercoralis in a young female tourist returning from Southeast Asia. To our knowledge, this is the first report of hookworm and Strongyloides stercoralis infection in a tourist from Europe, acquired in an endemic area.

Hydrogen peroxide-dependent antibacterial action of Melilotus albus honey.

Honey originating from different floral sources exhibits the broad spectrum of antibacterial activity as a result of the presence of hydrogen peroxide as well as nonperoxide bioactive compounds. The mechanisms of antibacterial activity of Polish melilot honey were investigated for the first time. Polish melilot honey samples (Melilotus albus biennial = 3 and annual = 5, Melilotus officinalis = 1) were collected directly from beekeepers and analysed for pollen profile, basic physicochemical parameters, antioxidant capacity, radical scavenging activity, total phenolic contents as well as antibacterial properties against pathogenic bacteria Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Salmonella spp. The physicochemical properties of melilot honey were specific for light-coloured unifloral honey samples and were not dependent on its botanical and geographical origin (P > 0·05). All tested honey samples exhibited inhibitory activity (above 90%) against Gram-positive bacteria at the concentration of 12·5-25%. Above 30-50% of antibacterial activity of melilot honey was connected with glucose oxidase enzyme action and was destroyed in the presence of catalase. Hydrogen peroxide-dependent antibacterial activity of honey was inversely correlated with its radical scavenging activity (r = -0·67) and phenolic compounds (r = -0·61). Antibacterial action of melilot honey depends not only on hydrogen peroxide produced by glucose oxidase, but also on other nonperoxide bioactive components of honey. SIGNIFICANCE AND IMPACT OF THE STUDY: Melilot honey is used in traditional medicine as an anticoagulant agent due to the possibility of the presence of the coumarin compounds which are specific for Melilotus plant. Melilotus albus is rarely used to produce honey, and antibacterial properties of this variety of honey had not been studied yet. Nine samples of melilot honey produced in different regions of Poland were analysed according to their antibacterial activity which was correlated with physiochemical parameters and antioxidant activity. It was shown that antibacterial activity of melilot honey is created by hydrogen peroxide and other bioactive compounds.

Molecular analysis of meticillin-resistant Staphylococcus aureus strains isolated from different types of infections from patients hospitalized in 12 regional, non-teaching hospitals in southern Poland.

BACKGROUND: Accumulated data indicate that meticillin-resistant Staphylococcus aureus (MRSA) infections are associated with a worse prognosis than methicillin-susceptible S. aureus infections. AIM: To assess the epidemiological profile of S. aureus infections and the genetic diversity of clinical strains of MRSA in 12 hospitals in southern Poland. METHODS: Samples from bloodstream infections, pneumonia, and skin and soft tissue infections from adult patients were examined. S. aureus isolates were tested for MRSA and macrolide-lincosamide-streptogramin B (MLSB) phenotypes. Staphylococcal cassette chromosome mec (SCCmec) typing and S. aureus protein A (spa) typing were performed. Analysis of the genetic similarity was performed by pulsed-field gel electrophoresis. RESULTS: This study included 555 patients. An MRSA phenotype was detected in 15.1% of strains. The prevalence of MRSA infection was higher in patients aged >80 years. An MLSB phenotype was detected in 18.2% of strains. Analysis of SmaI profiles did not reveal a dominant clone. Spa typing showed 25 different spa types, and spa type t003 was the most common (49% of strains). Among MRSA strains, SCCmecII (49%) and SCCmecIV (27.4%) were predominant. CONCLUSION: The characteristics of MRSA showed considerable heterogeneity. The results demonstrate the need for caution when drawing conclusions on direct epidemiological relationships between isolates based on a single typing method. As the cases of infection in this study were not associated with the hospital environment and horizontal transfer, a focus on screening at hospital admission, and appropriate infection control, may help to reduce the risk of MRSA infections.

The clinicopathologic significance of estrogen receptors in human gastric carcinoma.

In most populations, gastric cancer (GC) is approximately two times more prevalent in men than in women, which may suggest the protective role of sex steroid hormones in gastric carcinogenesis. Steroid hormones such as androgens and estrogens can be synthesized not only in the gonads but also in peripheral tissues. Many researchers have conducted studies examining the expression profile of enzymes involved in steroid synthesis, the occurrence of estrogen receptors (ERs) and the influence of ERs in the development, proliferation and progression of gastric cancer. Some studies have also evaluated the relationship between the presence of ERs and survival prognosis. However, the results of these studies are often controversial and divergent. In a recent study, it was indicated that sex steroid hormones and estrogen receptors are partly involved in gastric cancer but their clinicopathological significance still needs further investigation.

Prevalence and molecular characteristics of urinary and intestinal microsporidia infections in renal transplant recipients.

Transplant recipients have been identified as a new risk group for microsporidia infection. We characterize for the first time the prevalence of microsporidia in intestinal and urinary tracts of renal transplant recipients. Molecular examination of 86 patients showed that 25.5% of them were infected; 86% were confirmed to have pathogens in their urine and 45.5% in stool. Among positive patients, 32% had microsporidia confirmed in both urine and stool. Genotyping revealed Encephalitozoon cuniculi (59%) and Enterocytozoon bieneusi (23%) monoinfections as well as coinfections with both species (18%). Moreover, we found diarrhoea and fever as symptoms significantly associated with microsporidia presence. Our results indicate that microsporidial infection should be considered in the assessment of renal transplant recipients, especially in the urinary tract, even if asymptomatic. Molecular identification of microsporidia species is relevant because of their different susceptibility for treatment.

Urinary tract infections caused by Pseudomonas aeruginosa among children in Southern Poland: Virulence factors and antibiotic resistance.

OBJECTIVE: The aim of this study was to analyze antibiotic resistance and virulence patterns in Pseudomonas aeruginosa (PAR) isolates from urinary tract infections among children in Southern Poland. MATERIALS AND METHODS: This study comprised consecutive, non-repetitive PAR isolates sent from two collaborative laboratories. The study group consisted of children aged up to 17 years from Southern Poland with culture-proven PAR UTIs. Relevant information about patients with UTIs, such as age, sex, and type of infection (polymicrobial or monomicrobial), was collected. Isolates were screened for major virulence factors found in uropathogenic PAR strains. Multidrug-resistant (MDR) strains were defined as strains not susceptible to one antimicrobial in at least three different antimicrobial classes. Extensively drug resistant (XDR) strains were defined as strains susceptible to no more than two antimicrobial classes. RESULTS: The total prevalence of PAR UTIs was 2.1%, and in children <5 years of age it was 3.0%. A total of 26 isolates was tested: 21 from outpatients and five from inpatients. Most infections (80.8%) occurred in children ≤ 4 years of age. The most prevalent virulence gene was exoY (96.2%). The prevalence of other effector proteins was 88.5% for exoT, 92.3% for exoS, and 19.2% for exoU. The gene for LasB was present in 80.8% of isolates; the gene for AprA in 61.5%; the gene for PilA in 19.2%; and the gene for PilB was not detected. The PAR isolates were generally susceptible to beta-lactam and aminoglycoside antimicrobials. All isolates were also susceptible to colistin. A large proportion of isolates were resistant to carbapenems and fluoroquinolones (Fig. 1). No significant differences were found in antimicrobial resistance between males and females or inpatients and outpatients (p > 0.05 for all tested antimicrobials), or in antimicrobial resistance between younger (≤ 5 years old, n = 21) and older (> 5 years old, n = 5) children (p > 0.05 for all tested antimicrobials). Two isolates were classified as XDR and none as MDR. The EDTA test yielded one MBL-positive isolate (3.8%), from a 17-year-old patient in home care. No isolates with genes for the KPC, IMP, or VIM were identified. CONCLUSION: As data on UTIs in children with Pseudomonas etiology are scarce, this paper provides useful information for clinicians and allows for comparison between Poland and other countries. Our findings have important implications for clinicians treating UTIs empirically, because the success of empiric treatment is based on knowledge of pathogen antimicrobial susceptibility patterns.

Transplantation of keratinocyte spore-like stem cells.

BACKGROUND: Wound granulation tissue should be covered by epidermal cells migrating from the basal layer of the epidermis or hair "bulge" of the wound edge. However, new epidermal islands are frequently formed on the granulation tissue remote from the wound edge. Thus, current theory of "bulge"-originating stem cells does not necessarily correspond to the histological pictures of the healing wound. We took imprints of a leg ulcer surface and found single dispersed, large nucleated cells, some of them in mitosis. These cells resembled those from epidermal spinosum layer. The question arouse as to whether these cells might be the "spore-like" stem cells creating epidermal island. We found similarly shaped cells among the keratinocyte preserved in pulverized sodium chloride as the only surviving population in culture and revealing enzymatic activity. The aim of this work was to study whether the population of human keratinocytes surviving sodium chloride preservation and transplanted to SCID mice may form epidermis. METHODS: The 12-month sodium chloride-preserved and cultured keratinocytes (KC) were transplanted to the wound on the dorsum of SCID mice for 14 and 21 days. RESULTS: Ninety-five percent of cultured KC were enzymatically active "large" cells; they did not express p63 and CD29 claimed as specific for stem cells, and they did not proliferate. Transplanted to the center of the wound, they formed small KC islands and became confluent after 14 days. CONCLUSIONS: The "large" epidermal keratinocytes survived the 12-month preservation in anhydrous sodium chloride. Transplanted to the wound, they formed epidermal islands of human phenotype. These cells may be the so-called "spore-like" stem cells.

Skin own bacteria may aggravate inflammatory and occlusive changes in atherosclerotic arteries of lower limbs.

AIM: Seroepidemiological studies have given rise to the hypothesis that microorganisms like Chlamydia pneumoniae (CP), Helicobacter pylori (HP), cytomegalovirus (CMV), HCV types 1 and 2, and bacteria involved in dental or other unspecified infection sites may initiate or maintain the atherosclerotic process in lower limb arteries. However, not much attention has been attached to the patient's own limb skin and deep tissues bacterial flora, activated in ischemic tissues. This flora may enhance the inflammatory and thrombotic process in the atherosclerotic arteries. Lower limb tissues are exposed to microorganisms from the environment (foot) and microbes on floating epidermal cells from the perineal and anal regions. The aim of this paper was to identify microbial cells and their DNA in perivascular tissues and arterial walls of lower limbs. METHODS: Bacterial cultures and PCR method for detection of 16sRNA and immunohistopathological staining for identification of immune cells infiltrating vascular bundles. RESULTS: 1) specimens of atherosclerotic calf and femoral arteries contained bacterial isolates and/or their DNA, whereas, in control normal cadaveric organ donors' limb arteries or patients' carotid arteries and aorta bacteria they were detected only sporadically; 2) lower limb lymphatics contained bacterial cells in 76% of specimens, whereas controls only in 10%; 3) isolates from limb arteries and lymphatics belonged in majority to the coagulase-negative staphylococci and S.aureus, however, other highly pathogenic strains were also detected; 4) immunohistopathological evaluation arterial walls showed dense focal infiltrates of granulocytes and macrophages. CONCLUSION: Own bacterial isolates can be responsible for dense neutrophil and macrophage inflitrates of atherosclerotic walls and periarterial tissue in lower limbs and aggravate the ischemic changes.

Carrot infection by Alternaria radicina in field conditions and results of laboratory tests.

Carrot black rot caused by the fungus Alternaria radicina (Meier) Drechsler et Eddy, the economically important disease of carrot (Daucus carota L.). Research was carried out in 2004 and 2005 in order to establish correlation between plant infestation in the field and laboratory experiments. To determine the incidence and severity of the disease in mature crop, a 5-degree scale was used, where 1 meant no visible disease symptoms and 5 meant total foliage infestation. Plants were rated individually and the disease index was calculated. Fungus pathogenicity in laboratory conditions was determined using a petiole assay and a root disc assay. The petiole assay was conducted during the vegetation period; basal parts of petioles detached from fully developed carrot leaves were put in contact with the growing A. radicina mycelium. The root disc assay was conducted during storage of carrot roots; root discs were inoculated with toothpicks overgrown by A. radicina mycelium. The isolate of A. radicina was pathogenic to petioles and carrot discs. Differences in the reaction of the examined cultivars in the field and laboratory experiments were revealed. However, no significant correlation between plant infestation in the field and the results of laboratory experiments were observed.

Differentiation of cellular reaction to alloantigens and bacterial infection in human skin graft--immunosuppressive drugs or antibiotics.

The human hand transplantations prompted revival of interest in evaluation of the rejection process of the grafted skin and its control with the antirejection drugs [1-3]. In case of first hand transplantation a combined immunosuppressive regimen was applied with currently available drugs resulting in acceptance of the entire composite graft. No major untoward systemic effects of antirejection therapy were observed. The most important clinical conclusion was that allogeneic skin can be accepted and function as in a normal extremity, although the attack of host cells on the graft can not be totally eliminated. Chronic perivascular and subepidermal infiltrates with recipient cells could be seen [4]. Another problem connected with skin transplantation is graft infection. Skin is inhabited by a specific spectrum of bacteria [5]. Allografted skin is more sensitive to bacterial penetration than normal skin due to local damage by the host-versus-graft cellular reaction and compromised immune reactivity to bacterial antigens by the immunosuppressive therapy. The histological pictures of rejecting skin represent a mixture of cellular reaction against the graft and penetrating microbes. Alloreaction requires modification of immunosuppressive regimen and infection is an indication for prolonged antibiotic therapy against skin bacterial flora. The question arises how to discriminate the alloreactive and bacterial changes in the skin graft. We studied the histological pictures of rejecting and infected human skin after transplantion to scid mice.

Phenolic compounds in two Solidago L. species from in vitro culture.

The regenerated S. virgaurea and S. graminifolia, obtained previously by micropropagation from shoot tips and the callus tissue of S. virgaurea, were analysed for phenolic compounds. Preliminary used 2D-TLC methods (fingerprinting) showed the presence of phenolic glucoside--leiocarposide (S. virgaurea only), C-glycosyflavones--schaftoside and isoschaftoside (S. graminifolia), flavonoids and caffeoylquinic acids, similar to the ones reported earlier for naturally growing plants with the exception of the callus tissue. The latter contained predominantly caffeoylquinic acids, which were isolated and identified by NMR as 5-O-caffeoylquinic and 3,5-di-O-caffeoylquinic acids. The content of leiocarposide (HPLC) and flavonoids (colorimetry) in S. virgaurea was determined.

[The vestibular system and migraine in children]. / Uklad przedsionkowy a migrena u dzieci.

Migraine in childhood is relatively common disease. Although clinical practice and scientific publications recognize frequent association of migraine and vestibular disorders, relationships have yet to be well defined. In present study the vestibular function was extensively tested in 20 children suffering from migraine. All the patients underwent complete neurootological examination including tonal threshold audiometry, testing of vestibulo-spinal reflexes--Romberg and Unterberger-Fukuda tests, Dix-Halpike test for BPPV, videonystagmography. On videonystagmography each patient was evaluated regarding: calibration, spontaneous nystagmus, gaze nystagmus, eye tracking test, optokinetic and positional nystagmus, and caloric testing. The same procedure have been performed regarding control group comprised of 15 healthy. All the patients with migraine presented abnormalities in vestibular testing. Analysis of the results showed that: functional state of structures forming both visuo-ocular and vestibulo-ocular structures was altered in all the migraine patients evaluated in the study, pathological findings suggest mainly central localisation of vestibular dysfunction in children suffering from migraine, the number of pathological VNG findings seems not to be correlated with the type of migraine.

[Bird trematodes of lower Silesia. V. Trematodes of Diplostomatidae family parasitizing woodcock--Scolopax rusticola L]. / Przywry ptaków Dolnego Slaska V. Przywry z rodziny Diplostomatidae pasozytujace u slonki--Scolopax rusticola L.

Six dissections of Woodcock were conducted. Three birds were found to be infected by trematodes: Pulvinifer macrostomum and Scolopacitrema cubrensis (Diplostomatidae). Both of them are new species for the Polish parasite fauna. The examination of P. macrostomum has shown a greater variability in size of the body and organs than that described by other authors.

[Coxiella burnetii infections (Q fever) in animals and humans in Poznan and Leszno districts detected by serodiagnosis]. / Zakazenia Coxiella burnetii (Goraczka Q) u zwierszat i ludzi w województwach leszczynskim i poznanskim rozpoznane w badaniach serologicznych.

In Poznan and Leszno districts, serological survey of Q fever among cattle and sheep have been performed since 1973. Coxiella burnetii infections have not been detected until 1985. During this period 28,272 blood samples derived from cattle and sheep have been tested. For the first time in Poznan district, seropositive cattle were found in 1986 (5.5%). In the next year percentage increased to 16.3% and in following years has been on similar level. In Leszno district, serum antibodies to C. burnetii were detected in cattle in 1988 (25.9% seropositives). On the consecutive years, percentage of seropositive animals has decreased and in 1991 was 3.2% only. During these years there were 2.3 and 8.3% seropositive sheep detected in Poznan and Leszno districts respectively. Serological survey of C. burnetii infection has been performed among people exposed to this agent occupationally who undergo intermittent obligatory testing for brucellosis. In Leszno district among 4214 persons tested, 34.4% were seropositive. In Poznan district from 1988 to 1991 antibodies to C. burnetii have been detected in 22.7% of 6396 people tested. From year to year the percentage of seropositives fluctuated from 16.6 to 34.7%.

[Occurrence of infections with C burnetii (Q fever) in persons from increased risk groups in Poland--serological survey and evaluation of studied methods]. / Wystepowanie zakazen Coxiella burnetii (goraczka Q) u ludzi z grup zwiekszonego ryzyka na terenie Polski--przeglad serologiczny i ocena metod badawczych.

During years 1988-1991 at Regional Sanitary and Epidemiological Stations in Bialystok, Kielce, Lublin, Leszno, Piotrków Trybunalski, Lódz, Poznan, Plock, Opole, Sanok, Skierniewice and Zielona Góra--serological studies in persons amenable to compulsory check-up against brucellosis were performed for detection of infections with C. burnetii. In whole, 20651 persons were investigated on the Polish territory and it was found that in 22.8% antibodies against antigens of C. burnetii are present. Percentage of persons with detected antibodies varied in different regions from 0 to 41.7%. No antibodies were found in persons inhabiting regions of Kielce, Piotrków Trybunalski and Skierniewice. Most persons with antibodies indicating contact with C. burnetii were found in West Poland, namely in the Leszno and Poznan region. As in diagnosis of Q fever two methods (OWD and OMA) or OWD only were applied, it is difficult to compare results obtained in regions using two or only one method of determination.

Graft-induced polyclonal activation of B cells. I. Alloantigen-induced humoral response in mice.

In recent years evidence has accumulated pointing out that graft rejection may be mediated not only by cellular but also humoral mechanisms. To study this problem in some detail, humoral responses were studied in mice subjected to skin grafting across the H-2 locus. B cell reactivity was measured during the first and second set reactions on consecutive days after transplantation until graft rejection. In vitro B cell activity was assessed by means of the reverse plaque-forming cell assay (PFC) using cells obtained from the lymph nodes and spleens. The results indicate that polyclonal activation of B cells is an integral part of the immune response to skin transplantation in mice. However, no correlation was found between the level of activation as demonstrated by the PFC and the time of graft rejection.