Ear Samples Research Articles

Deciphering the mechanisms of the IL-6/CXCL1 and NOD-like receptor signaling pathways in otitis media with effusion in rodents

BackgroundOtitis media with effusion (OME) often leads to pediatric hearing loss and is influenced by innate and adaptive immune responses. Innate immunity serves as the non-specific first line of defense against OME. MethodsWe induced OME in rats using ovalbumin. We administered IL-6 monoclonal antibodies intranasally to inhibit IL-6, and we injected an NF-κB inhibitor intraperitoneally to explore the role of IL-6 in innate immunity and its interaction with the NOD-like receptor signaling pathway. We analyzed RNA-sequencing data with Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways to assess signaling pathways involved in OME. We also utilized Western blot, quantitative real-time PCR, and immunohistochemistry on middle ear samples and used microscopy to identify immune cells in ear wash fluids. ResultsOur study suggests a pivotal role for IL-6 in the immune pathways of rats with OME via the regulation of CXCL1-mediated pathways. Increased levels of IL-6 and CXCL1 were observed in the middle ear tissues, and activation of the NLRP3 inflammasome in OME rats led to an immune response via NF-κB, thus promoting IL-6 and CXCL1 production, which was reduced by IL-6 antibody treatment. ConclusionsOur findings confirm that IL-6 and CXCL1 play significant roles in the innate immune response in OME in rodents, predominantly via the NOD-like receptor signaling pathway and NLRP3 inflammasome activation. This research sheds light on OME pathogenesis and its immune-related mechanisms.

Evaluation of an in-house pan-Malassezia quantitative PCR in human clinical samples.

Althought Malassezia spp. have been involved in various pathologies, they are an integral part of the cutaneous, gut, oral, ears, nose and throat (ENT) mycobiota. Since Malassezia are difficult to grow in culture, unexhaustive molecular biology methods have been developed to detect them. The aim of the study was to evaluate an in-house pan-Malassezia quantitative polymerase chain reaction (panM-qPCR) on various clinical human samples and determine Malassezia burden in various human mycobiota. The panM-qPCR was designed to target the repeated 28S rDNA gene from all Malassezia species. We used the assay to quantify the Malassezia burden on 361 samples from 161 subjects (80 skin swabs from 10 healthy volunteers (HV), 13 samples from 2 seborrheic dermatitis patients (SD), 90 skin samples from 19 burned patients, 119 stool samples from 89 immunocompromised patients, 59 ENT samples from 41 patients). For HV, the amount of Malassezia was different according to the swabbed areas. Quantification cycle (Cq) in SD is lower than in HV. In burned patients, Cq was significantly lower compared to HV. In stool samples, 6.7% were positive for Malassezia spp. with a high Cq. For the ENT area, a higher proportion of positive specimens were detected in ear samples than in nose samples. Our findings emphasized the importance of qPCR, confirming elevated Malassezia spp. levels on individuals' faces and scalps, increased burden in SD patients and in severely burnt patients than in HV. The pan-MqPCR appears to be a promising tool for studying Malassezia in various human mycobiota.

Molecular and spatial evaluation of small rodents and Didelphimorphis infected with Mycobacterium leprae in the southern Amazon, Brazil.

The high levels of recent transmission of leprosy worldwide demonstrate the necessity of epidemiologic surveillance to understand and control its dissemination. Brazil remains the second in number of cases around the world, indicating active transmission of Mycobacterium leprae (M. leprae) in the population. At this moment, there is a consensus that the bacillus is transmitted by inter-human contact, however, different serologic, molecular, and histopathological approaches indicate the existence of non-human transmission sources. The qPCR assay was used to amplify the molecular targets 16S RNAr and RLEP, in samples of liver, spleen, and ear of wild animals belonging to Didelphimorphia and Rodentia orders, in highly endemic areas of Mato Grosso, Brazil. The RLEP repetitive sequence was positive in 202 (89.0%) samples, with 96 (42.3%) of these also being positive for the 16S gene. Regarding the collection sites, it was observed that the animals were found in areas profoundly deforested, close to urban areas. Our results suggest that wild animals can play an important role in the maintenance of M. leprae in endemic regions with major anthropic action in Brazil. Therefore, integrating human, animal, and environmental health care with the One Health initiative is highly efficient for the development of effective strategies to contain and control leprosy in Brazil.

Genome-wide association study indicates novel associations of annexin A13 to secretory and GAS2L2 with mucous otitis media

To evaluate the genetics of chronic nonsuppurative otitis media (OM). We performed a genome-wide association study of 429,599 individuals included in the FinnGen study using three different case definitions: combined chronic nonsuppurative OM (7034 cases) (included serous and mucous chronic OM), mucous chronic OM (5953 cases), and secretory chronic OM (1689 cases). Individuals without otitis media were used as controls (417,745 controls). We used immunohistochemistry (IHC) of the murine middle ear to evaluate the expression of annexin A13. Four loci were significantly associated (p < 1.7 × 10−8) with nonsuppurative OM. Three out of the four association signals included missense variants in genes that may play a role in otitis media pathobiology. According to our subtype-specific analyses, one novel locus, located near ANXA13, was associated with secretory OM. Three loci (near TNFRSF13B, GAS2L2, and TBX1) were associated with mucous OM. Immunohistochemistry of murine middle ear samples revealed annexin A13 expression at the apical pole of the Eustachian tube epithelium as well as variable intensity of the secretory cells of the glandular structure in proximity to the Eustachian tube. We demonstrated that secretory and mucous OM have distinct and shared genetic associations. The association of GAS2L2 with ciliary epithelium function and the pathogenesis of dysfunctional mucosa in mucous OM is suggested. The abundant expression of annexin A13 in the Eustachian tube epithelium, along with its role in apical transport for the binding and transfer of phospholipids, indicates the role of annexin A13 and phospholipids in Eustachian tube dysfunction.

Anti-inflammatory response to 1,8-Cineol and associated microbial communities in Otitis media patients

Chronic Otitis Media (COM) is defined as long term inflammation and colonization with pathogenic bacteria due to a defect or retraction of the tympanic membrane. Surgical interventions are often augmented by antibiotic resistance development and therefore, off-label treatment using the natural drug 1,8-Cineol was carried out. All COM patients underwent antibiotic therapy and middle ear surgery and developed antibiotic resistances. Microbiological investigations from the auditory canal and stool samples were performed in correlation with the clinical course. Therapy of COM patients with 1,8-Cineol revealed a clear reduction of inflammatory microbes P. aeruginosa and Proteus mirabilis in ear samples as well as intestinal Prevotella copri, which was associated with an improved clinical outcome in certain individuals. The present off-label study revealed manifold anti-inflammatory effects of the natural monoterpene 1,8-Cineol in Otitis media patients. A better understanding of the underlying mechanisms will improve the current treatment options and possible forms of application of this natural drug.

Prevalence of Mycobacterium leprae and Mycobacterium lepromatosis in roadkill armadillos in Brazil

To evaluate the prevalence of Mycobacterium leprae and Mycobacterium lepromatosis in road killed armadillos identified along Brazilian regions, samples of liver, spleen, muscle, ear, nose and tail were collected on highways from 78 animals. The armadillos were of four different species, Cabassous tatouay, Dasypus novemcinctus, Dasypus septemcinctus and Euphractus sexcinctus. After DNA extraction from two tissues, specific primers were used for the detection of each pathogen using SYBR green qualitative Real-Time PCR, and amplicons were sequenced. The species with the highest prevalence was D. novemcinctus, mainly in the Central-West, South, and Southeast regions of Brazil. We detected M. leprae DNA in 32 (41 %) of the 78 individuals and M. lepromatosis DNA was not identified in any of the examined samples. The zoonotic component of leprosy may play a role in the transmission of the disease in endemic areas in which environmental conditions and contact with reservoirs must be investigated.

Isolation and identification of Pseudomonas aeruginosa obtained from dogs and cats in Great Cairo regarding status of phenotypic antimicrobial resistance pattern

Background Companion animals; dog and cat are closely associated with the daily life of humans and may be implemented in the transmission of many microorganisms to their adopters. Pseudomonas aeruginosa (P. aeruginosa) is a suited opportunistic and harsh to treat pathogen due to its rife environmental distribution, unique intrinsic and acquired resistance to numerous antimicrobials’ categories. Objective The current study targeted to survey the existence of P. aeruginosa in laboratory samples obtained from diseased dogs and cats. The study also investigated the susceptibility and resistance of recovered isolates against antimicrobials. Materials and methods A total of 315 samples gathered from veterinary laboratories in Great Cairo governorates; fecal, ear, eye, respiratory, wounds and urine samples were previously collected from diseased dogs and cats. The samples were examined bacteriologically and biochemically to isolate P. aeruginosa. The isolates were assayed for their sensitivity and resistance versus 25 antimicrobials belonging to various categories. Results and conclusion Fifty- eight P. aeruginosa isolates (18.41%) were obtained from 315 dogs (44/233, 18.88%) and cats’ (14/82, 17.07%) clinical swabs. The isolates were confirmed biochemically and via VITEK 2 compact system. All isolates showed alpha-type of hemolysis and pigment production. The obtained P. aeruginosa isolates revealed a multidrug resistance pattern by 70.45% in dog isolates while cat isolates demonstrated a higher ratio 78.57%. P. aeruginosa isolates were highly resistant to cephalosporins, trimethoprim/sulfamethoxazole, and intermediate resistant to erythromycin fosfomycin. On the other hand imipenem, amikacin, azithromycin then gentamycin and ciprofloxacin were the most efficient on P. aeruginosa isolates. The study included that P. aeruginosa isolates obtained from canine and feline clinical samples collected from Great Cairo laboratories were characterized by high and intermediate levels of antimicrobial resistance. However, this pattern was directed to some classes of antibiotics, which are not authorized for veterinary use, which could expose an early warning mark and give the need for ongoing monitoring.

Morphological and molecular investigation of free-living amoeba Sappinia spp. isolated from environmental and clinical samples in Thi-Qar province-southern Iraq

Sappinia is a free-living amoeba, a single-celled living organism found in the environment. It has recently been attributed to being an opportunistic human pathogen. There are two known species of Sappinia: S. diploidea and S. pedata. The current study detected Sappinia species using morphological and genetic approaches in various environmental and clinical samples in the Thi-Qar province of southern Iraq. The study was conducted from February - September 2020; one hundred and two samples for this study were obtained from various environmental and clinical sources. PCR was carried out with a positive culture after the samples were cultivated on an NN-agar medium. Overall, the study showed that 53(51.96%) samples were positive on morphological characters as well as PCR analysis showed that only 47 (46.07%) ) of Sappinia morphologically positive samples were positive by using a specific primer. Sappinia spp. were observed in all types of samples of environmental and clinical sources except CSF, and clinical ear samples were negative. The current study showed two species of Sappinia that have similar morphology but differ in the diameter of trophozoite and cyst, number of nuclei, and cyst morphology. The current study will pave the way for additional epidemiological research by considering the presence of potential Sappinia species in various samples, whether from environmental or clinical sources, to better understand the function of Sappinia as a potential health danger to people. Keyword: Sappinia spp. Free-living amoeba, opportunistic amoeba, Thi-Qar, Iraq

Morphological and molecular investigation of free-living amoeba Sappinia spp. isolated from environmental and clinical samples in Thi-Qar province-southern Iraq

Sappinia is a free-living amoeba, a single-celled living organism found in the environment. It has recently been attributed to being an opportunistic human pathogen. There are two known species of Sappinia: S. diploidea and S. pedata. The current study detected Sappinia species using morphological and genetic approaches in various environmental and clinical samples in the Thi-Qar province of southern Iraq. The study was conducted from February - September 2020; one hundred and two samples for this study were obtained from various environmental and clinical sources. PCR was carried out with a positive culture after the samples were cultivated on an NN-agar medium. Overall, the study showed that 53(51.96%) samples were positive on morphological characters as well as PCR analysis showed that only 47 (46.07%) ) of Sappinia morphologically positive samples were positive by using a specific primer. Sappinia spp. were observed in all types of samples of environmental and clinical sources except CSF, and clinical ear samples were negative. The current study showed two species of Sappinia that have similar morphology but differ in the diameter of trophozoite and cyst, number of nuclei, and cyst morphology. The current study will pave the way for additional epidemiological research by considering the presence of potential Sappinia species in various samples, whether from environmental or clinical sources, to better understand the function of Sappinia as a potential health danger to people. Keyword: Sappinia spp. Free-living amoeba, opportunistic amoeba, Thi-Qar, Iraq

Effect of Fungicide Application and Corn Hybrid Class on the Presence of Fusarium graminearum and the Concentration of Deoxynivalenol in Ear and Stalk Parts of Corn (Zea mays) Used for Silage.

In Wisconsin, the use of brown midrib (BMR) corn (Zea mays) hybrids for ensiling and subsequent feeding to dairy cows is quite common. The overall milk production from cows fed silage from BMR hybrids is typically higher than those fed silage made from dual-purpose hybrids. Gibberella diseases (ear and stalk rot) caused by Gibberella zeae (anamorph; Fusarium graminearum) and the accompanying accumulation of the mycotoxin deoxynivalenol (DON) can be significant issues during the field production of BMR hybrids. The work presented here aimed to understand the role of hybrid class on the distribution of F. graminearum DNA and DON in the ear and stalk parts of corn for silage. An ear and stalk partitioned sample experiment was conducted on silage corn from field trials in Arlington, Wisconsin, in 2020 and 2021. The trials were arranged in a randomized complete block design in both years, including one BMR hybrid, one dual-purpose hybrid, and seven fungicide application regimes. Paired ear and stalk samples were physically separated, dried, and ground at harvest before determining the concentration of F. graminearum DNA and DON in each sample. Across both years, the main effects of hybrid, treatment, and plant part were not significant (P > 0.1) on DON concentration. However, the hybrid-by-plant part interaction effect was significant (P < 0.01). Ears of the BMR hybrid accumulated the most DON, whereas the dual-purpose hybrid ears had the lowest DON concentration. The concentrations of DON and F. graminearum DNA were significantly (P < 0.01) and highly correlated in the ear (r = 0.73) but not in the stalk (r = 0.09, P = 0.33). These findings suggest that DON accumulation in the corn ear is a major contributor in the difference observed in the total DON between the hybrid classes. Therefore, growers and researchers are encouraged to focus production and breeding on hybrids in both classes that accumulate less DON in ears, resulting in lower total DON in corn chopped for silage.

Anatomical Vascular Differences and Leishmania-Induced Vascular Morphological Changes Are Associated with a High Parasite Load in the Skin of Dogs Infected with Leishmania infantum.

Canine visceral leishmaniasis (CVL), caused by the protozoan Leishmania infantum, affects several organs, including the skin. Dogs are considered the major domestic reservoir animals for leishmaniasis, and through their highly parasitized skin, they can serve as a source of infection for sandfly vectors. Therefore, studies of the skin parasite-host relationship can contribute to the understanding of the infectious dissemination processes of parasites in the dermis and help to identify targets for diagnosis and treatment. Thus, the aim of this study was to evaluate the association of anatomical vascular differences and Leishmania-induced vascular morphological changes with clinical signs and parasite load by analyzing the ear and abdominal skin from dogs naturally infected with L. infantum. Paired samples of ear and abdominal skin from L. infantum-positive dogs (n = 26) were submitted for histological and immunohistochemistry analyses. The ear skin samples showed a more intense and more diffusely distributed granulomatous inflammatory reaction, a higher number and larger diameter of blood vessels, increased parasite load, higher expression of VEGF+ (vascular endothelial growth factor) and MAC 387+ (calprotectin) recently infiltrating cells, and more intense collagen disruption compared to the abdominal skin samples. Intracellular amastigotes were observed in blood vessels and inside endothelial cells and were diffusely distributed throughout the dermis in the ear skin samples. The NOS2/MAC387+ cell ratio was lower in the ear skin samples than in those of the abdomen, suggesting that in the ear dermis, the inflammatory infiltrate was less capable of producing NO and thereby control the parasite load. Together, these findings indicate how parasites and immune cells are distributed in the skin and suggest an important role for dermal vascularization in cellular influx and thereby in parasite dissemination through the skin of naturally infected dogs.

Bacteriological evaluation, antibiogram, and phenotypic detection of MBL-producing gram-negative bacteria isolated from outdoor patients with Otorrhea attending Ayub Medical Complex

Otitis media is frequently prevalent in developing countries and is primarily caused by bacteria. The current study aimed to study the patients to assess the occurrence rate, susceptibility pattern, and prevalence rate of metallo beta-lactamase (MBL) producing gram-negative bacteria to prevent the prevalent strains and direct the physicians to appropriate antibiotic therapy. The research study was conducted over nine-months, from November 2020 to July 2021, in which a total of 110 samples were collected from the outdoor-patients’ ear, nose, and throat (ENT) wards using sterile swabs and inoculated immediately on suitable culture media. The isolates were identified via conventional biochemical tests, and the susceptibility pattern of each isolate was tested using the Kirby-Bauer disc diffusion method. Out of the 110 total samples, merely 88.18% (97/110) ear samples were detected positive for the various strains of bacteria both in male and female patients with a mean age of 18.21 ± 15.86, contributing 54.64% and 45.36%, respectively. The most prevalent bacterial strain was Staphylococcus aureus (49.48%), followed by Pseudomonas aeruginosa (16.50%), Proteus spp. (13.40%), S. epidermidis (7.22%), E. coli (6.19%), S. saprophyticus (4.12%), and Klebsiella spp. (3.09%). Of the gram-negative isolated strains, only 18.75% (3/16) of the P. aeruginosa strains exhibited MBL production. A high prevalence rate was observed in the age-group 0–4 years (19.58%), followed by the age-groups, 5–9 and 15–19 (16.49% each), and the age-group >39 (10.31%). Levofloxacin, ciprofloxacin, amikacin, and fusidic acid were highly effective against the gram-positive, whereas ciprofloxacin, amikacin, and levofloxacin were potent against the gram-negative (except P. aeruginosa), while carbenicillin, meropenem and imipenem against the P. aeruginosa. The MBL-producers were highly (100%) resistant to IMP and MEM while highly (100%) sensitive to CAZ. In conclusion, the current study showed a high prevalence of S. aureus, P. aeruginosa, and MBL-producing P. aeruginosa (18.75%). Nevertheless, a high degree of resistance to antibiotics was noted, which could not be ignored.

Effect of diesel exhaust particles on RANK/RANKL expression in in vivo and in vitro models of middle ear inflammation

ObjectiveIncreasing evidence suggests a link between middle ear inflammation and the development of diesel exhaust particles (DEPs). Chronic middle ear inflammation can lead to bone damage and remodeling. This study aimed to explore the impact of DEPs on the expression of interleukin (IL)-6 and RANKL under conditions of middle ear inflammation. MethodsDEPs were collected by burning fuel in a diesel engine at the Gwangju Institute of Science and Technology. Human middle ear epithelial cells were cultured to 70–80% confluence in culture plates and then treated with DEPs at concentrations of 0, 5, 10, 20, 40, and 80 μg/mL for 24 h. Cell viability was assessed manually. B6.SJL mice, aged 9 weeks, were exposed to DEPs at a concentration of 200 μg/m3 for 1 h daily over a period of 28 days. The expression levels of IL-6, tumor necrosis factor α, RANKL, and RANK were evaluated using hematoxylin and eosin staining and western blot analysis of the harvested middle ear samples. ResultsThe viability of human middle ear epithelial cells was found to decrease in a dose-dependent manner after 24 h. The mRNA expression level of IL-6 exhibited the most significant increase at the 48-h mark. In contrast, the mRNA expression levels of RANKL and RANK showed a marked increase as early as 6 h post-exposure, with both genes subsequently displaying a time-dependent decrease. Histological analysis revealed that the middle ear mucosa was thicker in the group exposed to DEPs compared to the control group. Additionally, the protein expression levels of IL-6 and RANKL were elevated in the DEP-exposed group relative to the normal control group. ConclusionsWe confirmed the expression of osteoclast-related proteins in the mouse middle ear. These results imply that air pollutants might affect RANKL/RANK signaling, which is associated with bone remodeling.

Generation of allogenic chimera carrying mutations in PDX1 and TP53 genes via phytohemagglutinin-mediated blastomere aggregation in pigs.

The generation of genetically engineered pig models that develop pancreas-specific tumors has the potential to advance studies and our understanding of pancreatic cancer in humans. TP53 mutation causes organ-nonspecific cancers, and PDX1-knockout results in the loss of pancreas development. The aim of the present study was to generate a PDX1-knockout pig chimera carrying pancreas complemented by TP53 mutant cells via phytohemagglutinin (PHA)-mediated blastomere aggregation using PDX1 and TP53 mutant blastomeres, as a pig model for developing tumors in the pancreas with high frequency. First, the concentration and exposure time to PHA to achieve efficient blastomere aggregation were optimized. The results showed that using 300µg/mL PHA for 10min yielded the highest rates of chimeric blastocyst formation. Genotyping analysis of chimeric blastocysts derived from aggregated embryos using PDX1- and TP53-edited blastomere indicated that approximately 28.6% carried mutations in both target regions, while 14.3-21.4% carried mutations in one target. After the transfer of the chimeric blastocysts into one recipient, the recipient became pregnant with three fetuses. Deep sequencing analysis of the PDX1 and TP53 regions using ear and pancreas samples showed that one fetus carried mutations in both target genes, suggesting that the fetus was a chimera derived from embryo-aggregated PDX1 and TP53 mutant blastomeres. Two out of three fetuses carried only the PDX1 mutation, indicating that the fetuses developed from embryos not carrying TP53-edited blastomeres. The results of the present study could facilitate the further improvement and design of high-frequency developing pancreatic tumor models in pigs.

Molecular Analysis of Pseudomonas aeruginosa Isolates with Mutant gyrA Gene and Development of a New Ciprofloxacin Derivative for Antimicrobial Therapy.

This study focuses on the prevalence of Pseudomonas aeruginosa in various medical specimens. In addition, the investigates of this research shows the genetic analysis of pathogen-resistant isolates and chemical modifications to ciprofloxacin. A total of 225 specimens from men and women aged 30 to 60 were carefully collected and examined, including samples from wound, burn, urine, sputum, and ear samples. The data were obtained from AL Muthanna hospitals. PCR-RFLP and gene expression analysis were used to identify resistant strains and explore the genetic basis of antibiotic resistance. A ciprofloxacin derivative was synthesized and confirmed through FT-IR, 1H-NMR, and mass spectroscopy techniques then it was tested as antibacterial agent. Also, molecular docking study was conducted to predict the mechanism of action for the synthesized derivative. The results demonstrated that wound samples had the highest positive rate (33.7%) of P. aeruginosa isolates. The PCR-RFLP testing correlated ciprofloxacin resistance with gyrA gene mutation. Gene expression analysis revealed significant changes in the gyrA gene expression in comparison to the reference rpsL gene subsequent to exposure to the synthesized derivative. Furthermore, the molecular docking investigation illustrated the strategic positioning of the ciprofloxacin derivative within the DNA-binding site of the gyrA enzyme. The examination of genetic expression patterns manifested diverse effects attributed to the CIP derivative on P. aeruginosa, thus portraying it as a viable candidate in the quest for the development of novel antimicrobial agents. Ciprofloxacin derivative may offer new antimicrobial therapeutic options for treating Pseudomonas aeruginosa infections in wound specimens, addressing resistance and gyrA gene mutations.

Cloned Foal Born from Postmortem-Obtained Ear Sample Refrigerated for 5 Days Before Fibroblast Isolation and Decontamination of the Infected Monolayer Culture.

A 6-year-old mare, a valuable polo horse, died of complications following postcolic surgery. To preserve its genetics, ear skin samples were collected immediately after death and stored in an equine embryo transfer medium at 4°C for 5 days. After trypsin digestion, monolayer fibroblast cultures were established, but signs of massive bacterial infection were found in all of them. As an ultimate attempt for rescue, rigorously and repeatedly washed cells were individually cultured in all wells of four 96-well dishes. New monolayers were established from the few wells without contamination and used for somatic cell nuclear transfer. Four of the six Day 7 blastocysts derived from 14 reconstructed zygotes were transferred in four naturally cycling mares on Day 5 after ovulation. The embryo transfers resulted in 2 pregnancies, one from a fresh and one from a vitrified blastocyst. The vitrified embryo transfer resulted in a healthy offspring, now 21 months old, genetically and phenotypically identical to the somatic cell donor animal.

Clinical Uses of Nanopore Sequencing in Recurrent Acute and Chronic Otitis Media

Background. Otitis Media (OM) is a widespread problem that has an incidence of 10.85% worldwide, 51% occurring under the age of 5. Emergence of drug resistant bacteria contribute to high incidence of recurrent acute otitis media (AOM) and chronic otitis media with effusion (COME). Despite this, the standard of care for treatment is empiric antibiotics. Molecular microbiology diagnostics may enable clinicians to provided pathogen directed antibiosis. Nanopore sequencing is a highly accurate method of identifying a broad range of pathogens when compared with multiplex PCR. We hypothesize that nanopore sequencing is an effective method for detection of pathogens in the middle ear and nasopharynx of patients with recurrent or chronic OM.&#x0D; Methods. Bacterial Culture, multiplex PCR (BioFire PNA panel), and nanopore sequencing (MinIon, Oxford Nanopore Technologies) were tested on 60 middle ear and nasopharynx samples. Chi-Square test was used to examined differences in bacterial identification among the methods.&#x0D; Results. Bacterial identification using nanopore proves to be more sensitive in identification of 16 of the 18 unique pathogens when compared with multiplex testing. Of the 5 most common OM bacterial pathogens (S. pneumoniae, M. catarrhalis, H. influenza, S. aureus, P. aeruginosa), nanopore identified 3 pathogens (S. pneumoniae, M. catarrhalis, P. aeruginosa) at higher levels, 1 at the same rate (H. influenza), and 1 did not receive any readings (S. aureus).&#x0D; Conclusion. This is the first study to utilize nanopore sequencing to assess pathogens in the middle ear fluid of children with OM. While nanopore genomic sequencing is still in the early stages of use, it has potential to comprehensively identify bacteria with more sensitivity when compared to the current clinical standards. The next step of data analyzation will include bacterial identification in nanopore that multiplex PCR and traditional bacterial cultures cannot test for.

Comparative evaluation of macro- and microscopic changes in rabbit, cattle, and pig auricular cartilage following exhumation after different postmortem intervals

The study evaluated and compared macro- and microscopic changes in the auricular cartilage of rabbits, cattle, and pigs following exhumation at different postmortem intervals (PMI). Eight samples corresponding to 0, 5, 10, 15, 20, 25, 30, and 150 days after exhumation were obtained from all rabbit ears, and twelve samples corresponding to 0, 5, 10, 15, 20, 25, 30, 35, 40, 50, 60, and 150 days after exhumation were obtained from all cattle and pig ears. The weight loss of the rabbit ear samples remained higher than that of the cattle and pig ear samples at all PMIs. At 60 days, the average weight loss of cattle and pig ear samples did not differ significantly (p&gt;0.05). The loss of area of the samples was similar among the three species at most PMIs (p&gt;0.05). At 150 days, all cattle samples were completely decomposed, unlike the pig and rabbit samples. Microscopic analysis of the cartilage tinctorial affinity and loss of chondrocyte nuclei and tissue architecture demonstrated the worsening of postmortem changes over time, regardless of the animal species. Colonization by fungi and bacteria occurred earlier in the cattle and pig samples. The correlation of time with sample weight loss and cartilage thickness was strong, indicating the potential of these variables as parameters for PMI estimation. Therefore, postmortem examination of auricular cartilage can be used to estimate PMI.

ANALYSIS OF SOME BIOMETRIC PARAMETERS IN THE CHARACTERIZATION OF CORN EAR

The study analyzed the variation of some biometric parameters in corn ears, the 'Oana' hybrid. The experiment was carried out within ARDS Lovrin, under non-irrigated crop system conditions. The ear samples were harvested at physiological maturity. The analyzed parameters were represented by: ear length (EL), row number (RN), grains number on rows (GNR), ear weight (CW), grains weight (GW) and cob weight (CW). Ear length varied between EL = 18.00 – 20.75±0.41 cm; row number presented values between RN = 16.00 – 16.50±0.11; grains number on rows presented values between GNR = 43.75 – 49.25±0.76; ear weight presented the values EW = 0.86 – 1.04±0.03 kg; grains weight varied between GW = 0.73 – 0.87±0.03 kg; cob weight varied between CW = 0.13 - 0.17±0.01 kg. The Anova test confirmed the presence of variance and the statistical reliability of the recorded data (F calculated &gt; F crit, p&lt;0.001; Alpha = 0.001). High variability was recorded in the case of cob weight (CV=10.1184), and low variability in the case of row number (CV=1.5971). Intermediate values were recorded for the other analyzed parameters, respectively CV = 4.0769 for GNR, CV = 5.3064 for EL, CV = 7.6181 for GW, respectively CV = 7.9539 for EW. Correlation of varying intensity levels was recorded between analyzed parameters (e.g. r=0.991 between EW and GW; r=0.949 between EL and GNR; r=0.845 between CW and EW). The regression analysis facilitated the finding of equations and graphic models that described the variation of EW and GW in relation to EL and RN parameters, under statistical safety conditions (R2=0.982 in the case of EW; R2=0.974 in the case of GW). From the equations obtained (values of the coefficients) as well as from the graphic models, it was found that the EL parameter had a determining influence (compared to RN) in the variation of the EW and GW parameters.

Prediction of Ear Weight, Kernel Weight and Viability in Maize Using Image Analysis

In maize breeding studies, it is becoming common to determine the ear and kernel characteristics by image analysis. While current methods focus on measurements that can be obtained directly by image analysis, it has not been adequately addressed whether different parameters such as weight and viability can be estimated using these measurements. This study aimed to determine whether it is possible to estimate the ear weight (g), kernel weight (g), single kernel weight (g) and viability (1/0) status of maize with the help of features (area, perimeter, width, length) extracted from images of the ear and kernel samples. In this study, 233 ear and 1242 grain samples belonging to 13 maize genotypes were used as material. Digital images of the ear samples were taken with a 5 MP camera and from the kernel samples with a desktop scanner. The ear weight reference data (DV1) and the kernel weight reference data (DV2) were obtained by weighing each sample on a precision balance. Single kernel reference data (DV3) was obtained with the measurements of single kernel weights. Kernel samples underwent paper germination test and reference data (DV4) related to viability was created. Regression models were developed by using the features obtained from image analysis (area, perimeter, width, height) for each reference data set as the predictor variable. As a result of the study, it was seen that the ear weight and kernel weight can be estimated with the help of the parameters extracted from the image analysis. While moderate success was achieved in the determination of single seed weight, it was difficult to determine the viability status based on the morphometric measurements of a single kernel in maize.