The application of CLSI and EUCAST guidelines led to many discrepancies. Various doubts have already appeared in preliminary stages of microbiological diagnostics of Haemophilus spp. A total of 87 H. parainfluenzae isolates were obtained from throat or nasopharyngeal swabs from adults 18 to 70 years old, both healthy volunteers and patients with chronic diseases between 2013 to 2015 in eastern Poland. Haemophilus spp. were identified by colony morphology, Gram-staining, API NH and MALDI-TOF MS technique. Both susceptibility to various antimicrobials and phenotypes of Haemophilus spp. resistance to beta-lactams were determined. Statistically significant association between applied guidelines and drug resistance patterns were observed to as follows: ampicillin, cefuroxime, cefotaxime, amoxicillin-clavulanate, azithromycin, tetracycline and trimethoprim-sulfamethoxazole. Resistance phenotypes according to CLSI vs. EUCAST were as follows: 3.4% vs. 8.0% for BLNAR and 6.9% vs. 19.5% for BLPACR isolates. In conclusion, this is the first study that reports comparative analysis of drug susceptibility interpretation using CLSI and EUCAST of haemophili rods from human respiratory microbiota in Poland. In case of susceptible, increased exposure (formerly intermediate) category of susceptibility within H. parainfluenzae isolates we have observed EUCAST as more restrictive than CLSI. Moreover, BLNAI and BLPAI phenotype isolates have been observed, as well as BLPBR using only CLSI or EUCAST guidelines, respectively.

(1) Background: Beta-lactams are the most frequently used antimicrobials, and are the first-line drugs in many infectious diseases, e.g., pneumonia, otitis media. Due to this fact, various bacteria have developed resistance to this group of drugs. (2) Methods: Eighty-seven Haemophilus parainfluenzae isolates were obtained from adults 18–70 years old in eastern Poland. The presence of 10 bla genes and 2 substitutions in ftsI reported as the most frequent in H. parainfluenzae were analyzed. (3) Results: Among 57 beta-lactam-resistant isolates, 63.2% encoded bla genes; blaTEM-1 predominated (54.4%), followed by blaOXA (19.3%), blaDHA (12.3%), blaSHV (10.5%), blaGES (7.0%), blaCMY (5.3%), blaVEB (1.8%) and blaROB-1 (1.8%). Lys-526 was the most common substitution in ftsI gene. The resistance genotypes were as follows: gBLNAS (17.5%), low-gBLNAR I (1.8%), low-gBLNAR II (1.8%), gBLNAR II (15.8%), gBLPAS (15.8%), gBLPAR (19.3%), gBLPBS I (8.8%) and gBLPBS II (1.8%); (4) Conclusions: This has been the first study to report on the high diversity of bla genes in H. parainfluenzae isolates in Poland. High sensitivity and specificity of benzylpenicillin test, as well as PCR of bla genes were shown, indicating that these methods may be useful as tools for the rapid screening of beta-lactamase prevalence and resistance to beta-lactams among H. parainfluenzae isolated from respiratory microbiota.

Background. Haemophili are common human microbiota representatives. The aim of our study was to investigate a diversity of Haemophilus spp. isolates selected from clinical specimens on the basis of biochemical characteristics, biotypes distribution, protein profiles and antimicrobial resistance. Results. A total of 893/1025 (87%) of haemophili isolates were identified: 260/1025 (25%) as H. influenzae and 633/1025 (62%) as H. parainfluenzae. Moreover, a group of 107/1025 (10%) isolates without species identification (with e.g. abnormal numerical profile) was described as Haemophilus spp. Within the H. influenzae isolates, biotypes II and III were in a great majority (92/893; 10%, each), whereas among H. parainfluenzae, the most commonly occurring was biotype I and II (301/893, 34% and 178/893, 20%, respectively). A similar prevalence of biotypes was obtained regardless of the patient’s age or health condition or the type of specimen. A production of beta-lactamases was shown in 46/893 (5%) haemophili, both H. influenzae (13/46, 28%) and H. parainfluenzae (33/46, 72%) isolates. On the basis of haemophili biochemical characteristics, the cluster analysis using the UPGMA method demonstrated a high degree of phenotypic similarity due to a small distances between isolates taken from both unhealthy children and adults. Conclusion. Based on biochemical characteristics, about 90% of haemophili clinical isolates representing human-specific respiratory microbiota were positively identified as H. influenzae and H. parainfluenzae. The same differences in biotypes and antimicrobial resistance among isolates selected from healthy people or from patients with chronic and recurrent diseases were detected.

short duration of illness and an acute beginning. OME is the clinical term for the presence of fluid in the middle ear behind the intact eardrum and is characterized by a nonpurulent effusion that may be either mucoid (thick) or serous (thin) [4]. It is an inflammation of the middle ear without signs or symptoms of acute ear infection, pending beyond its intact eardrum and without systemic or otoscopic symptoms. An OME is the catchall term if the tympanic effusion persists for more than 3 months. Near chronic ARTICLE INFO ABSTRACT Introduction. Haemophilus influenzae and Haemophilus parainfluenzae are known as human-restricted respiratory microbiota representatives. The aim of the present paper was to assay haemophili prevalence in middle ear effusion specimens in pediatric patients with otitis media with effusion (OME). Methods. A total of 86 ear effusion specimens (from the left and right ear independently) were collected from 43 pediatric patients with OME. For comparison, 58 nasopharyngeal specimens were taken from 58 pediatric patients prone to recurrent respiratory tract infections (RRTI). Isolation and identification of haemophili biotypes and antimicrobial susceptibility was accomplished by standard microbiological methods. The cell surface hydrophobicity (CSH) of isolates was assayed by the method of aggregation in ammonium sulfate (SAT). Results. Haemophili were isolated in 25.6% (11/43) of all OME patients: in 5/43 (11.6%) -H. influenzae (biotypes II, III), in 5/43 (11.6%) -H. parainfluenzae, in 1/43 (2.3%) -both species were found. Haemophili-positive nasopharyngeal specimen was found in 27/58 (46.6%) RRTI patients: in 19/58 (32.8%) -H. influenzae, in 8/58 (13.8%) -H. parainfluenzae. About 90% of all haemophili isolates were characterised by extreme to strong CSH. Antimicrobial resistance occurred mainly among H. parainfluenzae (80%) and to a much lower percentage among H. influenzae (33.3%) isolates. The obtained data suggest that both H. influenzae and H. parainfluenzae can be involved in pathology of OME in pediatric patients. The high cell surface hydrophobicity can affect on the haemophili prevalence and ear colonization, and induces predisposition to the presence of these bacteria as a biofilm that serves as a virulence factor with great importance for the survival of these opportunistic bacteria and their persistence in the ear environment.

A most common infection is that the upper respiratory tract. The human body is inhabited by millions of microorganisms, most of which enable the proper functioning of some systems and constitute a barrier protecting against harmful external factors. Due to the continuous contact of the upper airway microbiota with the external environment, it may seem to be extremely unstable and very diverse. A number of studies have been conducted that have assessed the effect of various factors on the composition of the upper respiratory tract microbiota. This paper presents a literature analysis which showed that the anatomical area is rich in terms of the microbiota that forms it, and despite the constant changes in many parameters, including physical – very stable.