Culture Techniques Research Articles

The Role of Modified Mesenchymal Stem Cells on Endothelial and Stromal Cells in the Enhancement of Endometrial Injuries

Background: The uterine endometrium plays a important role in the processes of fertilization and embryogenesis, with its impairment or dysfunction leading to pathologies such as intrauterine adhesions, miscarriage, and infertility. In addressing endometrial damage, the application of stem cell has attracted considerable attention. To promote the paracrine capabilities of mesenchymal stem cells (MSCs), this study employed pro-inflammatory cytokines (Tumor Necrosis Factor-α and Interferon-γ, TNF-α and IFN-γ, IT) along with 3D culture techniques on pretreated MSCs (3D-IT-MSCs). We focused on evaluating the therapeutic potential of 3D-IT-MSCs and elucidating the mechanisms involved in endometrial repair. Method: Pretreated MSCs were co-cultured with human umbilical vein endothelial cells (HUVECs) or drug-induced endometrial stromal cells (ESCs) to observe the promoting effect on biological function. Results: The findings demonstrated that 3D-IT-MSCs exhibit markedly elevated paracrine molecule expression and secretion compared to conventional MSCs. Additionally, treatment with 3D-IT-MSCs significantly promoted the proliferation and migration of HUVECs and ESCs, resulting in increased HUVECs angiogenesis and inhibition of mifepristone-induced ESCs apoptosis. Conclusion: Our findings demonstrated that the combined approach of applying pro-inflammatory cytokines and 3D culture techniques on pretreated MSCs holds substantial promise as a therapeutic strategy for repairing endometrial injuries.

Using Microfluidic and Conventional Platforms to Evaluate the Effects of Lanthanides on Spheroid Formation

Metastasis contributes to the increased mortality rate of cancer, but the intricate mechanisms remain unclear. Cancer cells from a primary tumor invade nearby tissues and access the lymphatic or circulatory system. If these cells manage to survive and extravasate from the vasculature into distant tissues and ultimately adapt to survive, they will proliferate and facilitate malignant tumor formation. Traditional two-dimensional (2D) cell cultures offer a rapid and convenient method for validating the efficacy of anticancer drugs within a reasonable cost range, but their utility is limited because of tumors’ high heterogeneity in vivo and spatial complexities. Three-dimensional (3D) cell cultures that mimic the physiological conditions of cancer cells in vivo have gained considerable interest. In these cultures, cells assemble into spheroids through gravity, magnetic forces, or their low-adhesion to the plates. Although these approaches address some of the limitations of 2D cultures, they often require a considerable amount of time and cost. Therefore, this study aims to enhance the effectiveness of 3D culture techniques by using microfluidic systems to provide a high-throughput and sensitive pipeline for drug screening. Using these systems, we studied the effects of lanthanide elements, which have garnered interest in cancer treatment, on spheroid formation and cell spreading. Our findings suggest that these elements alter the compactness of cell spheroids and decrease cell mobility.

3D-printed microfluidic and millifluidic devices for cell culture and mechanotransduction studies: A review

The practice of conventional cell culture techniques affects the quality of biological research in mechanobiology by not replicating the cell’s microenvironment adequately. Microfluidic devices resembling 3D in vivo cell culture offer viable alternatives for mechanobiology studies and other applications including cell screening, cell separation, and point-of-care diagnostics. This review highlights recent advances in additive manufacturing (AM) to fabricate microfluidic and millifluidic devices through fast design iterations and cost reduction compared with conventional device manufacturing. Key advances in AM technologies such as fused deposition modeling (FDM), stereolithography apparatus (SLA), and digital light processing (DLP) have allowed the direct manufacture of complex microfluidic devices with master molds and sacrificial structures using multiple material components. Currently, available 3D printed devices for the precise fabrication of milli- and microfluidic devices, essential to mimicking the cellular microenvironment, while economically reducing production costs by eliminating expensive clean-room environments and reducing material waste are highlighted.

Evaluating the performance of traditional paddy (Oryza sativa L.) landraces under floating cultivation techniques in southern Tamil Nadu

Floating rice cultivation, a novel method, is essential for boosting crop yields and resilience amid climate change using flood-prone and water-saturated environments. This approach leverages aquatic resources such as lakes and reservoirs, enhancing agricultural productivity and ecosystem health. This study was conducted in 2023 at the Department of Agronomy, Agricultural College and Research Institute, Madurai to assess the suitability and production potential of cultivating traditional paddy varieties viz., Chinnar and Chithiraikar under floating conditions. Various substrates including soil, coir pith, wooden chips, sugarcane trash, thermocol, leaf compost, and jute bags were levelled out over paddy seeding trays (60 × 30 cm) and placed over bamboo rafts (120 × 120 cm) which served as the float base. Various parameters viz., germination (%), leaf area index (LAI), vigour index, plant population, dry matter production (g/m2), grain yield (kg/ha) and straw yield (kg/ha) were analysed. Results revealed that better crop establishment and crop growth were highly influenced by nursery and conditions with coconut coir outperforming other substrates. However, findings indicated that thermocol substrate is more productive in realizing higher yield attributes and yield (9.83 g/plant, 9.90 g/plant, respectively) of both Chinnar and Chithiraikar landraces of rice. Thus, floating rice cultivation offers a sustainable way to enhance productivity and adaptability, with traditional paddy varieties suitable for soilless media, though further research is needed to optimize its effectiveness.

Managing the parasitic honey bee mite Tropilaelaps mercedesae through combined cultural and chemical control methods

The western honey bee (Apis mellifera) is severely impacted by the parasitic Tropilaelaps mercedesae mite, which has the capacity to outcompete Varroa destructor mites (the current leading cause of colony losses) and more rapidly overwhelm colonies. While T. mercedesae is native to Asia, it has recently expanded its geographic range and has the potential to devastate beekeeping worldwide if introduced to new regions. Our research exploited the dependence of T. mercedesae on developing honey bees (brood) by combining a cultural technique (brood break) with U.S. registered chemical products (oxalic acid or formic acid) to manage T. mercedesae infestation. To evaluate this approach, we compared four treatment groups: (1) Brood Break; (2) Brood Break + Formic Acid (FormicPro®); (3) Brood Break + Oxalic Acid dribble (Api-Bioxal®); and (4) untreated Control. We found that the mite infestation rate of worker brood in Control colonies rose from 0.4 to 15.25% over 60 days, whereas all other treatment groups had infestation rates under 0.11% on Day 60. Mite fall assessments showed similar results, whereby Control colonies had 15.48 mites fall per 24 h on day 60 compared to less than 0.2 mites for any other treatment group. Evaluation of colony strength revealed that Brood Break + Formic Acid colonies had slightly reduced adult honey bee populations. No treatment eliminated all mites, so additional measures may be needed to eradicate T. mercedesae if detected in countries that do not currently have T. mercedesae.

Isolation and characterization of a resistance Bacillus subtilis for soil stabilization and dust alleviation purposes

Dust poses environmental, geological, health, and economic concerns, and microorganisms can help mitigate these adverse consequences by improving soil properties. Microbial calcium carbonate precipitation (MICP) has been found to be an efficient strategy for increasing soil strength, reducing soil porosity, and preventing erosion; however, severe environmental conditions such as pH and high temperatures may impede this process. To identify the best strain for MICP, 60 bacteria strains were obtained from arid soils using the enrichment culture technique. They were tested for the capacity of calcium carbonate deposition and biocement synthesis in stress environments. Phenotypic characterization indicated that the majority of the bacterial isolates were gram-positive and rod-shaped, with strong catalase and oxidase enzyme activity. Furthermore, MALDI-TOF MS identification revealed that the isolates were from the Bacillus and Pseudomonas genera. Scanning Electron Microscopy (SEM), Energy-dispersive X-ray spectroscopy (EDS), and X-ray diffraction (XRD) were used to analyze the microstructures and composition of bacterial cement. The results represented that B. subtilis isolate S56 has a higher production yield and forms distinctive calcite crystals as a result of fast urease synthesis. B. subtilis isolate S56 can be applied in situ to reduce soil erosion and dust pollution. This study reveals the potential of the B. subtilis S56 strain for soil consolidation and dust prevention in harsh environments and has the prospect of promoting its application in desertification control and ecological restoration.

Consumers’ perception and willingness to pay for hydroponic tomatoes: the effects of sustainability and quality attributes

PurposeThe agricultural sector is facing pressure due to concerns about its impact on the environment. Farmers must adapt to ensure high-quality, sustainable production. This requires efficient techniques such as soilless farming. The development of agricultural innovations depends on social acceptance; thus, it is crucial to identify the factors that influence consumers' purchasing decisions. The aim of this paper is to analyse consumers' perceptions of hydroponic cultivation techniques and their willingness to pay (WTP) a premium price for hydroponic tomatoes certified as “nickel-free” and “zero-residue”.Design/methodology/approachThe survey was conducted in Italy using tomatoes as a case study. Data were collected through an online questionnaire from a convenience sample of 292 respondents and were analysed using statistical analysis and a multiple linear regression model.FindingsThe results showed that WTP was influenced by frequency of purchase, familiarity with soilless technology, environmental sustainability, income and education. Consumers place a high value on the sustainability of the hydroponic production process and their perception of increased safety positively influences WTP. It is therefore recommended that marketing strategies focus on the environmental sustainability and safety of hydroponic products. In addition, it may be beneficial to implement a certification system specific to hydroponic cultivation, in addition to the existing “nickel-free” and “zero-residue” certifications.Originality/valueThis study introduces several novel elements: it is the first to assess the Italian consumers’ perceptions and WTP for a hydroponic product. Secondly, it assesses WTP in relation to several aspects of increasing relevance related to health claims, namely “nickel-free” and “zero-residue”.

Bacteriological Quality of Salads Sold at Selected Restaurants in Accra, Ghana

Background: The increasing prevalence of chronic non-communicable diseases has led to a greater emphasis on the consumption of healthy foods, such as vegetables. Vegetable salads from restaurants are generally perceived as safe. We investigated the bacteriological quality of vegetable salads sold in two popular restaurants in Accra. Methods: Twenty salad samples were purchased from two popular restaurants (A and B) with two branches each in Accra, Ghana. Restaurant A had branches at Dansoman and North Industrial Area, while B had branches at Osu and Tesano. Total aerobic colony forming unit (CFU) and biochemical assays were performed by standard culture techniques and protocols, to determine the microbial load and species present. Results: Mean aerobic bacteria count was 1.77E5 and 1.45 E5 CFU/g for Restaurants A, and B respectively. The North Industrial Area branch of A had more CFUs (2.64E5 CFU/g) than the Dansoman branch (0.9E5 CFU/g), and statistically significant (p=0.0010). The Tesano branch of restaurant B had higher CFUs (1.9E5 CFU/g) than the Osu branch (1.0E5 CFU/g), and also statistically significant (p=0.0022). Furthermore, ANOVA across the four branches showed a significant difference (p<0.0001). The main isolates identified from both restaurants were Enterobacter spp. (28.7%), Citrobacter spp. (20.4%), Klebsiella ssp. (18.5%) and Enterococcus spp. (7.4%). Conclusion: Enterobacter species was predominant among others. Education of the restaurant staff, and the application of food safety and handling procedures must be established, and food regulatory institutions must carry out routine inspection at these sites to ensure consumer protection and public health.

Efficacy of Plant Tissue Culture Techniques for Eliminating Black Mulberry Idaeovirus (BMIV) from Infected Black Mulberry (Morus nigra)

Obtaining virus-free plants is a crucial step in disease management that enables reliable and profitable fruit farming. The present study applied various in vitro virus elimination protocols, including apical shoot culture, chemotherapy, thermotherapy, cryotherapy, and their combination, to eliminate black mulberry Idaeovirus (BMIV) from sour black mulberry. First, a shoot tip (0.5–2 mm) culture protocol was optimized, and four ribavirin concentrations (0, 10, 20, and 30 mg/L) were investigated over five weeks as a form of chemotherapy (ch). For the first thermotherapy treatment (Ch + Th1st), chemotherapy treatment was followed by a gradual increase in the temperature (24–33 °C). In another experiment (Th2nd + Ch), in vitro shoots were incubated in the dark for two weeks at two different temperatures (35 ± 1 °C and 37 ± 1 °C, for one week each). Subsequently, the shoot tips were incubated with various ribavirin doses. Finally, cryotherapy (Cr) was used with or without immersing the shoot tips in liquid nitrogen. A two-step RT-PCR was performed to assess the presence of the virus in 7–8-week-old in vitro plants. Th2nd + Ch significantly increased the shoot tip burst and plant survival/morphogenesis compared to the other treatments. Except for the application of cryotherapy, the protocols eliminated BMIV in different proportions, and the highest virus elimination rate (50%) was obtained by applying 30 mg/L ribavirin during the Ch + Th1st treatment. These findings are essential in preventing the dissemination of the virus and enabling the safe movement of germplasm around the world.

Interspecific hybridization and cell engineering of lettuce (Lactuca L.)

Lactuca sativa L. is a leafy vegetable crop of the Asteraceae family, widely cultivated throughout the world. The main breeding trends for lettuce include higher yields, better taste quality, earliness, and resistance to abiotic and biotic stressors. Some wild Lactuca spp. have actively been employed by lettuce breeders as donors of resistance to various diseases. Conventional and biotechnological breeding methods are both currently used to develop new lettuce cultivars. This is an overview of the main advances in the production of interspecific Lactuca hybrids, including the use of cell and tissue culture techniques, and genetic engineering. Studying artificial hybridization and natural populations makes it possible to identify evolutionary relationships among various Lactuca spp. Somatic hybridization is an overlooked but promising technology in Lactuca breeding: it allows a breeder to obtain a wider range of variations, and is beyond the strict control by GMO laws. This technique faces problems associated with complicated protoplast regeneration and the loss of reproductive ability in hybrids. Genome-editing methods are more effective and better controllable, but society is still wary of any interference with the plant genome and legally regulates the sale of GM products as food. Thus, researchers are challenged with the task to improve these techniques.

Unveiling the Human Brain on a Chip: An Odyssey to Reconstitute Neuronal Ensembles and Explore Plausible Applications in Neuroscience.

The brain is an incredibly complex structure that consists of millions of neural networks. In developmental and cellular neuroscience, probing the highly complex dynamics of the brain remains a challenge. Furthermore, deciphering how several cues can influence neuronal growth and its interactions with different brain cell types (such as astrocytes and microglia) is also a formidable task. Traditional in vitro macroscopic cell culture techniques offer simple and straightforward methods. However, they often fall short of providing insights into the complex phenomena of neuronal network formation and the relevant microenvironments. To circumvent the drawbacks of conventional cell culture methods, recent advancements in the development of microfluidic device-based microplatforms have emerged as promising alternatives. Microfluidic devices enable precise spatiotemporal control over compartmentalized cell cultures. This feature facilitates researchers in reconstituting the intricacies of the neuronal cytoarchitecture within a regulated environment. Therefore, in this review, we focus primarily on modeling neuronal development in a microfluidic device and the various strategies that researchers have adopted to mimic neurogenesis on a chip. Additionally, we have presented an overview of the application of brain-on-chip models for the recapitulation of the blood-brain barrier and neurodegenerative diseases, followed by subsequent high-throughput drug screening. These lab-on-a-chip technologies have tremendous potential to mimic the brain on a chip, providing valuable insights into fundamental brain processes. The brain-on-chip models will also serve as innovative platforms for developing novel neurotherapeutics to address several neurological disorders.

Characterization of middle ear microbiome in otitis media with effusion in Hungarian children: Alloiococcus otitidis may potentially hamper the microbial diversity

IntroductionAlthough otitis media with effusion (OME) is a common cause of hearing impairment in children, little is known about its microbiological background. We aimed to analyse the microbiome of the middle ear fluid (MEF) in OME to identify microbiological, demographic and environmental factors that may potentially influence the middle ear microbiome. In addition, we aimed to compare the results of conventional culture techniques and PCR-based methods. Methods39 samples from children with OME were investigated using conventional culture and 16S rRNA sequencing. Bioinformatic analyses were carried out to assess the microbial communities and their association with clinical data. ResultsBy conventional culture technique bacteria could be cultured in 33 % of the MEF samples; the most frequent bacterium was Haemophilus influenzae, followed by Staphylococcus species, Cutibacterium acne, and Streptococcus species. The 16S rRNA analysis showed that Alloiococcus was the most abundant bacterium, followed by Haemophilus, Streptococcus. and Sphingobium.A “High Alloiococcus group” with significantly lower and a “Low Alloiococcus group” with significantly higher alpha and beta diversity could be defined. Children born by Caesarean sections had lower beta and Shannon diversity than patients born by natural birth. Breastfeeding for at least six months showed a negative correlation with alpha diversity. Age, previous antibiotic treatment, parental smoking, duration of symptoms, existence of siblings, did not alter the significantly bacterial composition of MEF samples and there were no significant differences in regard to alpha or beta diversity. ConclusionThe conventional culture technique and 16S rRNA results were not congruent. Remarkably, Alloiococcus could not be cultured at all, even by 16S rRNA analysis the presence of Alloiococcus seems to be important. Based on our results, A. otitidis possibly impacts the diversity of middle ear microbiome in children with OME. However, further studies are required to determine the role of A. otidis in middle ear pathologies.

Chilgoza Pine: A Comprehensive Review on Ecology, Cultivation, and Economic Potential of an Underutilized Nut Crop

Chilgoza pine (Pinus gerardiana L.) is an underutilized nut crop native to the high-altitude regions of South Asia, particularly the western Himalayas. This comprehensive review aims to provide an in-depth understanding of Chilgoza pine's ecology, cultivation practices, and economic potential. The review synthesizes current knowledge on the tree’s ecological requirements, including its adaptation to arid and mountainous environments. It examines cultivation techniques, challenges faced in agronomic practices, and recent advancements in improving yield and quality. Additionally, the economic potential of Chilgoza pine is explored, highlighting its role in local economies, market trends, and opportunities for commercialization. This review underscores the significance of Chilgoza pine in sustainable agriculture and its potential as a high-value nut crop.

The super-intensive culture of Penaeus vannamei in low salinity water: A comparative study among recirculating aquaculture system, biofloc, and synbiotic systems

The culture of Penaeus vannamei in inland regions using low salinity water is a reality in several regions of the world. The dissemination of culture techniques that allow high stocking density and reduced water use can be used to optimize the production process, but they require comparative studies to better understand the functioning of these systems in those salinity conditions. Therefore, this study aimed to analyze the effects of different culture systems, which were recirculating aquaculture system (RAS), biofloc technology (BFT), and synbiotic system (Synbiotic) on the water quality, plankton composition, and growth of P. vannamei in low salinity (2 g L−1) and high stocking density (500 shrimp m−3) for 30 days. The shrimp were stocked at a mean weight of 1.27 ± 0.06 g. In the BFT, dextrose was used as the organic carbon source and administered at a carbon: nitrogen ratio of 15:1. In the Synbiotic, rice bran processed by probiotic microorganisms was used as an organic fertilization strategy. In the RAS treatment, all nitrogen species remained stable throughout the trial, without spikes, and with ammonia and nitrate concentrations lower than in BFT and Synbiotic. The BFT treatment had more events when total ammonia nitrogen exceeded 1 mg L−1, requiring water changes. The Synbiotic treatment had better control of nitrogenous compounds, and a more accentuated accumulation of nitrate compared to BFT, suggesting more effective nitrification. The BFT treatment had more microalgae than Synbiotic and RAS. However, the abundance of zooplankton was higher than that of phytoplankton in BFT and Synbiotic, indicating a dominance by heterotrophic organisms. The Synbiotic treatment had a higher abundance of ciliates and amoeba than the other treatments. Furthermore, the Synbiotic treatment provided higher survival, yield, and lower FCR than BFT and RAS. Our findings indicate that the Synbiotic system can be considered as an alternative for the P. vannamei super-intensive culture in low salinity water, as it presented better control of nitrogenous compounds compared to BFT, higher abundance of ciliates and amoeba, and provided better production rates to the culture.

Hybrid Coffee (<i>Coffea arabica</i> L) Plantlet Production via Indirect Somatic Embryogenesis in Ethiopia: Current Statue and Future Direction

Traditional hybrid coffee propagation methods using seed or vegetative cuttings has tremendous limitations. Seed propagation is associated with hand pollination which is time consuming and need large number of skilled labor. On the other hand, vegetative cuttings ensure uniformity, but; cuttings generate relatively low multiplication rates as they can only be obtained from orthotropic branches. Multiplication by tissue culture techniques could provide a best alternative to these traditional methods of coffee propagation as it allowed the production of relatively uniform plantlets on a large scale in a shorter period in any climatic condition. Among tissue culture techniques, indirect somatic embryogenesis of hybrid coffee plays substantial role in rapid industrial scale multiplication of high valued varities. Research on somatic embryogenesis of <i>C. arabica</i> hybrid has been conducted since the late 70s with the objectives to substitute the conventional vegetative propagation of selected varieties. In Ethiopia, in vitro propagation of hybrid coffee is recent phenomenon and research on indirect somatic embryogenesis is already well underway by different research groups. Here, we discussed the current status of coffee research on somatic embryogenesis in general and specifically, we provide recommendations for future research for the establishment of mass propagation protocol for F1 hybrids varities in Ethiopia that utilized wild endogenous lines.

Managing the Nutraceutical and Sensorial Qualities of Pisanello, an Ancient Tomato Landrace, in Soilless Conditions

Recently, there has been significant consumer demand for traditional tomato varieties due to their favourable organoleptic qualities; however, the cultivation of these ancient varieties is becoming more restricted due to inadequate shelf life and low productivity. The “Pisanello” is a Tuscany tomato variety mainly cultivated in the provinces of Pisa, Lucca, and Livorno, and the main producers of this ancient tomato are small local farmers. The purpose of this work was, firstly, to study the range of quality parameters of this landrace tomato grown using different cultivation techniques, both in soil and soilless systems. For this purpose, the physicochemical parameters of Pisanello tomatoes grown in six different farms in Tuscany using both soilless and soil methods were investigated. Secondly, Pisanello tomatoes grown using different soilless techniques (rockwool and aeroponics) and soil-grown tomatoes (Pisanello and Goldmar F1) were evaluated from organoleptic and nutraceutical points of view. The sensory profile evaluation of all types of tomatoes under investigation was carried out. The aeroponic cultivation of Pisanello induced higher organoleptic qualities than those of tomatoes cultivated in rockwool (+34% for titratable acidity and +18% for total soluble solids). On the other hand, soilless rockwool-grown tomatoes showed a better sensory profile with respect to aeroponic cultivation. Nevertheless, the Goldmar F1 tomato, morphologically similar to ‘Pisanello’, received lower scores from the sensory panel compared to the Tuscany landrace tomato. This indicates that ancient tomato varieties selected over decades remain the preferred choice for consumers. Therefore, from a long-term viewpoint, the valorisation of local tomato varieties such as Pisanello can promote the regional commercialization of novel niche products originating from ancient fruit thanks to their acceptability by consumers.

Mesenchymal stromal cells as cancer promoters.

Mesenchymal stromal cells (MSCs) are important cellular constituents of tumor stroma that play an active role in tumor development. Complex interactions between MSCs and cancer promote tumor progression by creating a favorable milieu for tumor cell proliferation, angiogenesis, motility, invasion, and metastasis. The cellular heterogeneity, source of origin, diversity in isolation methods, culture techniques and model systems of MSCs, together with the different tumor subtypes, add to the complexity of MSC-tumor interactions. In this review, we discuss the mechanisms of MSC-mediated tumor promotion and evaluate cell-stromal interactions between cancer cells, MSCs, cells of the tumor microenvironment (TME), and the extracellular matrix (ECM). A more thorough understanding of tumor-MSC interactions is likely to lead to better cancer management.

Crucial Factors Influencing the Efficiency of Androgenesis in Oat (Avena sativa L.) Through Anther and Microspore Cultures

Historically, traditional crossbreeding schemes have predominated in oat breeding. In vitro culture techniques seek to expedite the breeding process and enhance selection efficiency. Maximum yields are achieved from hybrid plants produced by crossing pure (homozygous) lines with the desired traits. Homozygous lines can be produced through conventional breeding methods, which are time-consuming and costly. Alternatively, the production of homozygous lines can be accelerated by producing doubled haploid (DH) plants derived from (haploid) male gametophytes or their microspores (androgenesis). This method condenses the various stages required for producing homozygous lines in a single generation, resulting in significant time and cost savings. These and other advantages render androgenic DHs the preferred choice in numerous important crops where any of the various in vitro experimental techniques (anthers culture or isolated microspores culture) are well-established. However, in the case of oat (Avena sativa L.), an efficient plant regeneration method remains not very effective compared to the most common cereals, possibly due to the known recalcitrance of this cereal to in vitro culture. This review presents the methods through anther and microspore cultures utilized in the production of oat DHs revealing the crucial factors influencing the efficiency of this method in oat (Avena sativa L.).

Simultaneous determination of somatic cell count and total plate count in raw milk based on ATP bioluminescence assay

The somatic cell count (SCC) and total plate count (TPC) are essential quality indicators for raw milk. Traditional detection methods require separate measurements and rely on complex, large-scale instruments or cultivation techniques, which are both time-consuming and laborious. To address these challenges, this study developed a novel method for the simultaneous detection of SCC and TPC in the same raw milk sample using the ATP bioluminescence assay. This method utilizes oxy-ethylated iso-nonyl phenol (Neonol-10) and cetyltrimethylammonium bromide (CTAB) to selectively lyse somatic cells and microorganisms, respectively. This technique is straightforward to operate and can be completed within 2.5 h, with detection ranges of 1 × 10⁴ to 3 × 10⁶ cells/mL for SCC and 1 × 10⁵ to 5 × 10⁷ CFU/mL for TPC. Importantly, this technique meets the requirements of detection standards in China, European Union, Canada, United States, etc. For SCC or TPC in raw milk. Overall, this innovative approach does not rely on expensive equipment or facilities and the stepwise reagent addition procedure can be easily developed into an automated high-throughput system for rapid on-site testing of SCC and TPC in raw milk.

Development of an Effective Sterilization Protocol for Plant Tissue Culture Studies in Superfruit Aronia [Aronia melanocarpa (Michaux) Elliot

Effective sterilization protocols are crucial for a successful tissue culture study in Aronia. These protocols directly influence contamination rates, shoot health, and root development. In this context, the aim of the study is to develop an effective sterilization protocol for plant tissue culture studies in Aronia [Aronia melanocarpa (Michaux) Elliot], commonly known as the "superfruit." In study, shoot tips of the Nero Aronia variety were used as material. The sterilized shoot tips were transferred to the respective plant tissue culture media in a randomized parcels trial pattern with three replicates, each containing three explants per replicate. Various concentrations and combinations of sterilizing agents, such as sodium hypochlorite (NaOCl), hydrogen peroxide (H2O2), mercuric chloride (HgCl2), and ethanol (C2H5OH), were evaluated to determine their effectiveness in maintaining tissue health and reducing contamination. A total of twelve protocols were developed, incorporating different concentrations of these chemicals. The results showed that the combination of 5% NaOCl and 3% H2O2 provided the lowest average contamination rate (5%) and the highest average number of healthy (uncontaminated) explants (9.00 piece), demonstrating the sterilization efficiency of this combination. On the other hand, protocols containing HgCl2, especially at higher concentrations, resulted in impaired root development. High ethanol concentrations also contributed to effective sterilization, with the combination of 7% NaOCl and 80% ethanol yielding a low contamination rate (8%) and preserving tissue health. This study emphasizes the need to balance sterilization protocols between effective contamination control and tissue viability. The findings are expected to benefit the improvement and development of tissue culture techniques for Aronia and similar species, providing a basis for further research on effective sterilization practices, which are currently limited in Aronia tissue culture.