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Avian pathogenic E. coli (APEC), produces an extraintestinal infection in chickens, turkeys, and other types of birds, called colibacillosis, which is considered one of the main causes of economic losses due to morbidity, mortality, and discard of poultry carcasses. The objective of the present study was to characterize the genetic profile of the virulence factors of different isolates of avian E. coli in Caloto, Cauca, Colombia. Materials and methods: E. coli was isolated and identified by biochemical tests, from 47 clinical isolates. Subsequently, the DNA was extracted using Chelex. Three multiplex PCRs were designed to amplify 13 virulence factors (iroN, hlyF, iss, iutA, frz, vat, sitA, KpsM, sitD, fimH, pstB, sopB, and uvrY), using primers previously reported for each. At the end, the amplification products were verified on agarose gels. Each isolate was classified according to the number of virulence factors: group A (between 10 and 13), group B (between 5 and 9), and group C (4 or less). Discussion and Conclusions: we were able to identify the presence of a group of virulence factors in clinical isolates of APEC, which allows us to demonstrate that both the frequency and the profile of virulence factors in the isolated strains showed a different profile than the reported by other authors. The virulence genes pstB and fimH were detected in all our samples, and the iss gene was the one with the lowest frequency. Finally, according to the number of virulence factors, the group A was the most frequent.

Millions of patients worldwide suffer from gastrointestinal (GI) motility disorders such as gastroparesis. These disorders typically include debilitating symptoms, such as chronic nausea and vomiting. As no cures are currently available, clinical care is limited to symptom management, while the underlying causes of impaired GI motility remain unaddressed. The efficient movement of contents through the GI tract is facilitated by peristalsis. These rhythmic slow waves of GI muscle contraction are mediated by several cell types, including smooth muscle cells, enteric neurons, telocytes, and specialised gut pacemaker cells called interstitial cells of Cajal (ICC). As ICC dysfunction or loss has been implicated in several GI motility disorders, ICC represent a potentially valuable therapeutic target. Due to their availability, murine ICC have been extensively studied at the molecular level using both normal and diseased GI tissue. In contrast, relatively little is known about the biology of human ICC or their involvement in GI disease pathogenesis. Here, we demonstrate human gastric tissue as a source of primary human cells with ICC phenotype. Further characterisation of these cells will provide new insights into human GI biology, with the potential for developing novel therapies to address the fundamental causes of GI dysmotility.

Background & objectives: Trypanosoma cruzi and Leishmania spp. are protozoans that cause American trypanosomiasis and leishmaniasis, respectively. In endemic foci where both diseases coincide, coinfection can occur. The objective of this work was the characterization of the parasites involved in coinfection in several endemic areas of Venezuela. Methods: Molecular characterization was done in 30 samples of several species of mammals (Didelphis marsupialis, Equus mulus, Rattus rattus, Canis familiaris, Felis catus, and Sciurus granatensis) from the states of Anzoategui, Cojedes and Capital District diagnosed with T. cruzi and Leishmania spp. coinfections. For the typing of T. cruzi DTUs, the markers of miniexon, 24Sa rDNA, 18Sa rDNA, and hsp60-PCR-RFLP (EcoRV) were used. Infection by Leishmania spp. was characterized by miniexon multiplex PCR for complexes of Leishmania and ITS1-PCR-RFLP (HaeIII, HhaI, and RsaI) for the identification of the species. Results: The T. cruzi TcI was present in 100% of the coinfected mammals, which included 76.7% of triple infection by T. cruzi TcI-complex-L. (L) mexicana-L. infantum/chagasi, 13.3% of double infection by T. cruzi TcI-L. mexicana and 10% of double infection by T. cruzi Tcl-L. infantum/chagasi. Interpretation & conclusion: These results suggest that the double or triple infection is a phenomenon existing in almost all the coendemics areas and mammals studied, which might influence the mechanisms of adaptation and pathogenicity of these parasites.

Background & objectives: For detection and molecular characterization of Babesia microti in laboratory mice from India. Methods: A total of 625 mice were screened by peripheral blood smear examination and subsequently was confirmed by PCR using a piroplasm conserved primer set (Piro A/B). Nested PCR was done using a species-specific primer targeting the gene encoding the small subunit ribosomal RNA (18S rRNA). The PCR products were cloned, purified and sequenced. A total of 12 isolates were obtained. The sequences were aligned and phylogenetic trees were prepared with other published Babesia spp. sequences. Results: B. microti was detected with a total infection rate of 8.80%. The higher rate of infection was observed by species specific PCR (8.80%) than examined by blood smear (7.20%). Sequence and phylogenetic analysis showed that Babesia species detected in mice were genetically identical to the genotypes of B. microti and can be easily distinguished from other genotypes of Babesia parasites by neighbour joining and maximum likelihood method. Intra-species analysis indicated that all the twelve isolates from six North-Eastern states of India have a close identity but inter-species showed genetic reservoir host for transmission of babesial infection to humans. Interpretation & conclusion: The detection of Babesia microti may suggest that laboratory mice may serve as potential reservoir host for human infection and possibility of innovative way of diagnosing and control of human babesiosis.

Olfactory neuroblastomas are uncommon malignancies that arise from olfactory receptor cells located high in the nasal cavity. Accurate diagnosis plays a crucial role in determining clinical results and guiding treatment decisions. Diagnosis can be a major challenge for pathologists, especially when dealing with tumours with poor differentiation. The discovery of several molecular and immunohistochemical markers would help to overcome classification difficulties. Due to the paucity of large-scale studies, standardisation of diagnosis, treatment and prediction of outcome remains a challenge. Surgical resection by endoscopic techniques with the addition of postoperative irradiation is the treatment of choice. In addition, it is advisable to consider elective neck irradiation to minimise the risk of nodal recurrence. Molecular characterisation will help not only to make more accurate diagnoses but also to identify specific molecular targets that can be used to develop personalised treatment options tailored to each patient. The present review aims to summarise the current state of knowledge on histopathological diagnosis, the molecular biology and management of this disease.

Gastrodia elata Blume is an important medicinal orchid, yet its large-scale cultivation is increasingly threatened by fungal diseases. The 4-coumarate–CoA ligase (4CL) gene family directs a key step in phenylpropanoid metabolism and plant defence, but its composition and function in G. elata have not been investigated. We mined the G. elata genome for 4CL homologues, mapped their chromosomal locations, and analysed their gene structures, conserved motifs, phylogenetic relationships, promoter cis-elements and codon usage bias. Publicly available transcriptomes were used to examine tissue-specific expression and responses to fungal infection. Subcellular localisation of selected proteins was verified by transient expression in Arabidopsis protoplasts. Fourteen Ge4CL genes were identified and grouped into three clades. Two members, Ge4CL2 and Ge4CL5, were strongly upregulated in tubers challenged with fungal pathogens. Ge4CL2 localised to the nucleus, whereas Ge4CL5 localised to both the nucleus and the cytoplasm. Codon usage analysis suggested that Escherichia coli and Oryza sativa are suitable heterologous hosts for Ge4CL expression. This study provides the first genome-wide catalogue of 4CL genes in G. elata and suggests that Ge4CL2 and Ge4CL5 may participate in antifungal defence, although functional confirmation is still required. The dataset furnishes a foundation for functional characterisation and the molecular breeding of disease-resistant G. elata cultivars.

is the prevalent parasitic protozoan responsible for diarrhoeal disease in humans and animals. This study aimed to determine the prevalence and assemblages of this parasite present in cattle in Poland. Faecal samples were collected from 1,602 cattle up to 4 months old, bred on 267 farms located in all 16 voivodeships of Poland. Extraction of DNA was performed using a modified alkaline and a heat lysis method. Molecular detection of gene by PCR and sequence analysis were performed. In 174 of 1,602 (10.9%) tested cattle and in 89 of 267 examined herds (33.3%), DNA was detected. The highest prevalence of in cattle was found in the Lubelskie (25%), Mazowieckie (21.3%) and Zachodniopomorskie (20.4%) voivodeships. In other regions, the prevalence did not exceed 10%. The number of -positive cattle decreased with animal age. The most frequently identified assemblage in cattle was E (89.2%), and A and B were detected more seldom in 9.0% and 1.8% of cattle, respectively. No significant differences in positive results were observed in cattle depending on production purpose. The study results showed a high prevalence of in cattle in Poland. The presence of the A and B assemblages indicates a potential zoonotic threat.

ABSTRACT A total of 178 microbial cultures were isolated from rhizospheric soil, of which 45 cultures were identified as actinomycetes based on morphology. After primary and secondary screening, the actinomycete GK-121 was found to be a potent biocontrol agent for the rice blast pathogen Pyricularia oryzae. The culture was identified using conventional microscopy, Scanning Electron Microscope (SEM) and molecular characterization of the 16S rRNA sequence, and showed 99% similarity with Streptomyces maritimus, which was further confirmed by phylogenetic analysis. The culture exhibited a 45 mm zone of inhibition during in vitro screening against P. oryzae. The greenhouse experiment further proved the effectiveness of the isolate, as the seeds treated with S. maritimus showed very low disease incidence in plants inoculated with the pathogen P. oryzae, whereas untreated seeds showed high disease incidence. This is the first report to suggest that S. maritimus is effective in controlling the rice blast pathogen, P. oryzae, in vitro and in vivo. Keywords: Rhizosphere, Actinomycetes, Antiphytopathogenic activity, Molecular characterization, Biocontrol

Dissolved organic matter (DOM) plays a fundamental role in nutrient cycling dynamics in riverine systems. Recent research has confirmed that the concentration of riverine DOM is not the only factor regulating its functional significance; the need to define the chemical composition of DOM is a priority. Past studies of riverine DOM rested on bulk quantification, however technological advancements have meant there has been a shift towards analytical methods which allow the characterisation of DOM either at compound class or more recently molecular level. However, it is important to consider that all analytical methods only consider a defined analytical window. Thus, herein, we explore the use of a hierarchy of methods which can be used in combination for the investigation of a wide range of DOM chemistries. By using these methods to investigate the DOM composition of a range of streams draining catchments of contrasting environmental character, a wide range of compounds were identified across a range of polarities and molecular weight, thereby extending the analytical window. Through the elucidation of the DOM character in stream samples, information can be collected about likely the sources of DOM. The identification of individual key compounds within the DOM pool is a key step in the design of robust and informative bioassay experiments, used to understand in-stream ecosystem responses. This is critical if we are to assess the role of DOM as a bioavailable nutrient resource and/or ecotoxicological factor in freshwater.

Biogenic amines (BAs) constitute an important chemical hazard in fish and fishery products, primarily due to their adverse effects on human health. While histamine-producing bacteria have been extensively investigated, the contribution of Enterococcus species to BA formation in fresh marine fish remains poorly characterized. The present study aimed to isolate and identify Enterococcus spp. from fresh anchovy (Engraulis encrasicolus) and to evaluate their capacity to produce major BAs. A total of 56 fresh anchovy samples were analyzed, yielding 24 Enterococcus isolates that were characterized by phenotypic methods and 16S rRNA gene sequencing. The BA production capacities of the isolates were quantified via HPLC following incubation in a standardized liquid screening medium under optimized laboratory conditions. Fourteen isolates (58.33%) exhibited aminogenic activity and were identified as Enterococcus faecalis (42.86%), E. faecium (28.57%), E. lactis (21.43%), and E. durans (7.14%). Tyramine, histamine, tryptamine, cadaverine, and putrescine were detected, with maximum concentrations reaching 307.638, 257.939, 236.928, 165.835, and 56.317 mg/L, respectively. The highest total BA levels, E. faecium BLK18, exhibited the highest levels, reaching 493.547 mg/L. These findings demonstrate that fresh anchovies may serve as a reservoir for aminogenic Enterococcus spp. and highlight their potential role in BA accumulation beyond traditionally implicated bacteria. The results support the inclusion of Enterococcus species in routine monitoring programs for fresh fish products and underline the need for broader, evidence-based strategies to manage BA-related hazards in seafood.