Agar Medium Research Articles

Establishment of an In Vitro Micropropagation Protocol for Hibiscus moscheutos L. ‘Berry Awesome’

Hibiscus moscheutos L. ‘Berry Awesome’ is a complex hybrid of the new Proven Winners Summerific series of varieties with highly ornamental characteristics. Micropropagation of highly ornamental varieties is important for mass production of planting material for commercial purposes. The traditional methods for propagating Hibiscus varieties, such as cuttings or seed propagation, however, do not guarantee high rates of production of high-quality seedlings. To solve this problem, an attempt was made to develop protocols for micropropagation of Hibiscus moscheutos L. ‘Berry Awesome’ in vitro on agar and liquid medium using a bioreactor system, followed by ex vitro adaptation of the regenerants. The optimal method for sterilization of nodal explants as well as the optimal composition of the initiation medium for shoot proliferation and rooting were determined. For micropropagation on a liquid medium, a rocker-type bioreactor was used, and its advantages over micropropagation on an agar medium were demonstrated. The results showed that the best sterilization method for nodal segment explants was as follows: pretreatment by rinsing with running tap water, sterile water, and distilled water for 70 min and soaking for 5 min in a mixture of solutions of ethyl alcohol (96%), hydrogen peroxide (38%), and water in a ratio of 1:1:2. In this case, live and sterile explants accounted for 62.6%. The optimal initiation medium for axillary buds in nodal segments was the Murashige and Skoog (MS) medium supplemented with 0.1 mg L−1 N-(2-chloro-4-pyridinyl)-N’-phenylurea (CPPU), which resulted in 73.3% of axillary buds being induced. The optimal solid proliferation medium was MS medium supplemented with 0.1 mg L−1 CPPU with a proliferation coefficient of 5.8. In a liquid medium, the optimal concentration of CPPU was 0.05 mg L−1 with a proliferation coefficient of 9.2. The best medium for rooting/shoots with agar and in bioreactors was MS medium with the addition of 0.1 mg L−1 indole-3-butyric acid (IBA). The highest rooting rate was 99.0% in both types of media, and the survival rate of plantlets was 88.7% in solid media and 98.7% in the bioreactor.

Races and Biovar Characterization of Ralstonia solanacearum Causing Bacterial Wilt Disease of Potato

Ralstonia solanacearum, the causative agent of brown rot/bacterial wilt, is a major cause of severe potato illnesses in many impoverished nations located in tropical and subtropical regions of the world. When it occurs, the disease is to blame for significant losses to the potato industry. The illness has the potential to completely destroy a crop and stop using a piece of land for potato cultivation for a number of years. A survey was carried out to investigate the extent of Ralstonia solanacearum-caused bacterial wilt of potato crops in important potato-growing talukas in the Banaskantha district of Gujarat, India, including Deesa, Dantiwada, Palanpur, and Amirgadh. The frequency of bacterial wilt disease in potatoes is rather low in this region. Four talukas were utilised to identify ten isolates (RsSt1 to RsSt10) of R. solanacearum, which were then used to determine the biovar and race. Gram-negative rod-shaped cells were present in the isolates of R. solanacearum. On TZC agar medium, these isolates developed colonies that were creamy or dull white in colour with a hint of pink or red in the centre. The oxidation of disaccharides (sucrose, lactose, maltose) and sugar alcohols (manitol, sorbitol, and dulcitol) by R. solanacearum isolates was used to determine the biovar characteristics. This revealed that, of the ten isolates, eight were classified as bv2, and the two remaining isolates (RsSt5 and RsSt7) belonged to bv2T. The pathogenicity test conducted on tobacco, tomato, and brinjal allowed for the race identification of R. solanacearum isolates, which were classified as race 3 due to their limited host range and ability to exclusively elicit wilt symptoms in potatoes and tomatoes. As a result, Races 3 and bv2 and bv2T were represented among the R. solanacearum isolates that caused bacterial wilt of potatoes in the Banaskantha district.

INHIBITORY POWER OF MAS BANANA PEEL EXTRACT (MUSA ACUMINATA LADY FINGER) ON THE GROWTH OF STAPHYLOCOCCUS AUREUS IN VITRO

Introduction: Various sources of infectious disease can be found in the oral cavity. Prevention efforts include brushing your teeth properly and regularly. Besides that, gargling with mouthwash is one of the best additional prevention methods. In the literature it is known that mas banana peels contain antibacterial ingredients. The aim of this research was to determine whether banana peel extract (Musa acuminata lady finger) was able to inhibit the growth of Staphylococcus aureus bacteria. Material and Methods: Extracts of 25%, 50%, 75% concentration of dried mas banana peel were made, sterile distilled water as a negative control and 0.2% Chlorhexidine as a positive control. Staphylococcus aureus ATCC 25923 as many as 3 bacterial colonies, spread evenly on sterile Nutrient Agar (NA) media. The paper discs were dipped for ± 30 seconds into 25%, 50%, 75% gold banana peel extract, 0.2% Chlorhexidine as a positive control and sterile Aquades as a negative control then incubated in an incubator at 37°C for 24 hours. The inhibition zone (clear area around the paper disc) was measured with a caliper (accuracy 0.02mm). The data obtained were tested for normality and homogeneity to determine whether or not there was a difference in the inhibitory power of banana mas (Musa acuminata lady finger) peel extract against Staphylococcus aureus bacteria. Results and Discussion: The results showed that at a concentration of 75% there was an inhibitory power for the growth of Staphylococcus aureus bacteria of 6.32 mm. Mas banana peel extract (Musa acuminata lady finger) has antibacterial content in the form of flavonoids, alkaloids, saponins and tannins which are able to inhibit the growth of Staphylococcus aureus bacteria in vitro. Conclusion: Mas banana peel extract (Musa acuminata lady finger) can inhibit the growth of Staphylococcus aureus bacteria in vitro with moderate criteria.

THE EFFECTIVENESS OF CEMCEM (Spondiaspinnata (L.f) Kurz) LEAF EXTRACT IN INHIBITING THE GROWTH OF Streptococcus mutans

Introduction: Dry socket is post-tooth extraction pain characterized by partial or complete loss of blood clots in the alveolar socket. One of the management of dry socket is by using antibiotic therapy. Inappropriate use of antibiotics in addition to being economically wasteful can also cause bacterial resistance to antibiotics. The use of herbal ingredients such as Cemcem (Spondiaspinnata (L.f) Kurz) is one solution. Cemcem leaves are thought to have antibacterial effects because they contain flavonoids, tannins and saponins. This study aims to determine the inhibition of cemcem leaf extract (Spondias pinnata (L.f) Kurz) against the growth of Streptococcus mutans. Materials and methods: The research method used is the Kirby Bauer method which is a sensitivity test with the agar diffusion method using the disc diffusion technique. In the Kirby Bauer sensitivity test using selective media, namely Muller Hinton Blood Agar media, the inhibition zone created is characterized by a clear zone around the disc. The data obtained were analyzed using the Kruskall Wallis test to determine whether or not there was a significant difference then continued with the Mann Whitney U - Test. Results: Cemcem leaf extract (Spondias pinnata (L.f) Kurz) with a concentration of 60% and 70% can inhibit the growth of Streptococcus mutans better than positive control and 80% concentration. Conclusion: 60% concentration of cemcem leaf extract is more effective in inhibiting the growth of Streptococcus mutans than 70% and 80% concentrations.

Symptomatology and Morpho-Cultural Characterization of Colletotrichum musae, a Causal Agent of Anthracnose Disease in Banana

The study focused on the identification and characterization of Colletotrichum musae as a significant post-harvest fungal pathogen causing anthracnose disease in bananas. Symptoms included sunken, water-soaked spots on infected tissues, leading to soft, red-brown to black-coloured spots with irregular shapes. The pathogen was isolated and cultured on Potato dextrose agar medium, and its pathogenicity was confirmed by inoculating healthy banana fruits, resulting in typical anthracnose-like spots. Cultural characteristics of C. musae on PDA revealed whitish to black colonies, and microscopic examination showed oblong to cylindrical, single-celled conidia. Acervuli, which were salmon-coloured initially and later turned dark brown to black, were observed on the lesions. The fungus exhibited maximum growth and sporulation on PDA, followed by oat meal agar and Richard’s agar. Additionally, the study highlighted the morphological features of C. musae, including septate, irregularly branched, and vacuolated mycelium. The findings contribute to understanding the pathology and characteristics of C. musae, providing valuable information for managing post-harvest anthracnose in bananas.

ANTIBIOTIC SUSCEPTIBILITY PROFILE AND NUTRITIONAL EVALUATION OF RAW AND COOKED MEAT OF CYPRINUS CARPIO

The Cyprinus carpio fish belongs to the family Cyprinidae and nutritionally consists of beneficial amino acids and omega-3 polyunsaturated fatty acids. However, consuming fish contaminated with pathogenic bacteria causes serious human health issues. The study aimed to identify pathogenic bacteria, antibiogram profile and extended-spectrum-beta-lactamase (ESBL) producing bacteria, and nutritional composition of C. carpio fish in Peshawar, Pakistan. Fish samples (30 uncooked and 30 cooked) were collected from Peshawar and analyzed in the laboratory to detect pathogenic bacteria. The bacteria were isolated, and different sorts of staining and biochemical tests including Catalase, Oxidase, Triple sugar iron, Urease, Citrate utilization, Indole etc,. were performed for identification. Antibiotic susceptibility pattern was tested on Muller Hinton agar medium (MHA) against nine antibiotics. Nutritional analysis of raw and cooked meat was performed using the Association of Official Analytical Chemists procedure. The results showed that E. coli has the highest resistance (100%) towards Tetracycline and Penicillin, while K. pneumonia showed the highest antibiotic resistance (100%) against Tetracycline, Oxacillin, Ampicillin, and Penicillin. Sheigilla was observed to the highest resistance against Penicillin and Oxacillin, which was 100%, and Salmonella showed the highest resistivity (100%) towards Tetracycline, Oxacillin, and Penicillin. Similarly, Tetracycline, Oxacillin, and Ampicillin showed the highest antibiotic resistance (100%) toward S. aureus. In 60 samples, 105 bacterial species (E. coli, Salmonella, Sheigella, Klebsiella, and S. aureus) were isolated. All isolated bacteria were ESBL producing except S. aureus. The results showed that crude fat (3.5-10.4%), crude fiber (0.23-0.38%), crude ash (5.6-5.7%), crude protein (69.03-89%), and moisture (4.6-5.8%) were greater in uncooked fish meat as compared to cooked fish meat. The study concludes that the high prevalence of pathogenic bacteria in uncooked meat is observed compared to cooked meat of C. carpio. Uncooked fish meat is more nutritious than cooked fish meat as the amount of protein, fats, fiber, ash, and moisture decreases during cooking.

Homofermentative Lactobacilli isolated from organic sources exhibit potential ability of lactic acid production.

There has been an increasing interest in recent years in lactic acid bacteria that are derived from organic sources for lactic acid production. This research article presents the isolation and identification of homofermentative lactic acid bacteria from various novel organic sources, followed by qualitative and quantitative analyses of lactic acid produced. A total of 32 isolates were identified initially from various sources, such as curd (C1, C2), probiotics (P1, P2, and P3), silage (Si1 and Si2), soil samples (S1, S2, and S3), vermicompost (V1 and V2), and Farmyard manure. Biochemical tests such as Gram's staining, catalase test, and oxidase test were conducted for preliminary identification of lactic acid bacteria using De Man, Rogosa, and Sharpe agar (MRS) media. Through selection and identification, based on colony morphology and biochemical characteristics, 18 isolates were identified as lactic acid bacteria. The subsequent analysis included a tube test, screening for organic acid production, and homofermentative screening using homofermentative-heterofermentative differential (HHD) medium for qualitative analysis of lactic acid. The results revealed that 9 out of 18 selected strains were homofermentative and had promising potential for the production of lactic acid. Furthermore, six isolates (P1-1, S1-3, C2-1, V2-3, P2-1, and C1-1) from all of the nine positive strains were subjected to pH testing (0, 24, 48, and 72 h) and titrimetric assay for estimation of % crude lactic acid present. The presence of lactic acid was confirmed using thin-layer chromatography (TLC). L (+)-Lactic acid was quantified using a K-LATE enzymatic assay kit, for the best three isolates (P1-1, S1-3, C2-1). Finally, the strains were subjected to 16SrRNA sequencing and were identified as Lactobacilli. Based on the findings of the study, it could be concluded that homofermentative lactic acid bacteria with significant LA-producing ability can be obtained from different organic sources and may prove to be useful in the successful production of lactic acid for biotechnological applications.

The HEMOLYSIS ACTIVITY OF BACTERIA ISOLATS FROM PELANGI FOREST OF IJEN GEOPARK

Ijen geopark is one of Indonesia's geoparks, which is located in East Java. A total of 153 bacteria have been isolated from Pelangi Forest, which were then given the isolate code IHP. These bacterial isolates can be utilized in industries, including organic matter decomposer agents, plant biocontrol agents, and probiotics. To ensure these bacteria are safe to use in various fields, they must be non-disease-causing (non-pathogenic). Safety evaluations based on the hemolysis reactions offer simple tests to ease the analysis of potential pathogenic bacteria. The study aimed to evaluate the safety of bacterial isolates from Pelangi Forest for their hemolysis reactions. The hemolysis test was conducted using blood agar media, from which isolates with a negative (λ) reaction. Based on the results of hemolysis tests, 30 out of 153 bacterial isolates (19.60%) were found to be negative reactions. These bacteria are safe to proceed with for potential analysis.

KELIMPAHAN DAN KARAKTERISTIK BAKTERI DALAM BAKASANG BIA GARU (Tridacna gigas L) [Abundance and Characteristics of Bacteria in Bakasang Bia Garu (Tridacna gigas)

Bakasang bia garu is a clam-based fermented product processed by the people of the Babar Islands, Maluku. Uncontrolled processing allows the presence of unwanted bacteria in these local food products. Preservatives added in the processing of bakasang bia garu are very diverse. This study aims to isolate and characterize lactic acid, halotolerant, and coliform bacteria from bakasang bia garu which are processed using different preservatives. Lactic acid bacteria were isolated using de Man Rogosa and Sharpe Agar (MRSA) media while halotolerant and coliform bacteria used Mannitol Salt Agar (MSA) and Eosin Methylene Blue Agar (EMBA) media, respectively. Characterization was carried out macroscopically and microscopically. The results showed that lactic acid bacteria were only found in bakasang with synthetic salt and vinegar preservatives of 3.5x103 CFU/g which was dominated by Pediococcus pentosaceous. Halotolerant and coliform bacteria have diverse characteristics that belong to the genera Staphylococcus, Escherichia, and Enterobacter. Bakasang bia garu needs to optimize its controlled processing to improve the quality of this local food product in the future

Properties of Potential Plant-Growth-Promoting Bacteria and Their Effect on Wheat Growth Promotion (Triticum aestivum) and Soil Characteristics

Plant-growth-promoting bacteria are an important economic and environmental resource as biofertilizers that can stimulate plant growth and improve agricultural yields. In this study, potential plant growth-promoting bacteria were isolated from soil samples collected in Russia. Strains that manifested active growth on a nitrogen-free medium, the Pikovskaya medium (with insoluble phosphates) and CAS (Chrome Azurol S) agar, were selected for the study. All bacterial isolates were identified by 16S rRNA gene sequencing analysis. Seventeen bacterial isolates of different species were purified and quantified for their ability to grow on nitrogen-free media; dissolve phosphate; and produce ammonium, indole-3-acetic acid, siderophores, and antifungal activity. Principal component analysis identified three groups of strains: one with the maximum signs of providing “plant nutrition”; one with signs of “antimicrobial activity”; and a group “without outstanding signs”. All 17 strains were involved in experiments involving growing inoculated wheat seeds (Triticum aestivum) in pots under natural environmental conditions, and were assessed by their effect on the wheat growth and yield as well as on the chemical composition of the soil. For the “plant nutrition” group, regression analysis revealed a connection between indicators of plant growth, ear length, and ammonium accumulation in the soil. However, in other groups, there were also strains showing a positive effect on plant growth, which suggests the necessity of involving additional factors to predict the ability of strains to affect plants when screened in the laboratory.

The study on the impact of sex on the structure of gut microbiota of bamboo rats in China.

Bamboo rats are rodents that eat bamboo, and their robust capacity for bamboo digestion is directly correlated with their gut flora. Chinese bamboo rat (Rhizomys sinensis) is a common bamboo rat in Chinese central and southern regions. As a single-stomach mammal, bamboo rats are a famous specificity bamboo-eating animal and their intestinal microbial composition may also play a key role in the digestion of cellulose and lignin. So, the gut microbiota of bamboo rat may play an important role in the adaptation of bamboo rats for digesting lignocellulose-based diet. To study the microbiome differences of bamboo rats from different sexes, the microbial genomic DNA was extracted from each fecal sample and the V4 region of 16S rRNA genes was amplified and sequencing on an IlluminaHiSeq6000 platform. The operational taxonomic units (OTUs) were classified, the OTUs in different sexes was identified and compared at phylum and genus levels. For isolation and screening of cellulose degradation bacteria from bamboo rats, fresh feces from randomly selected bamboo rats were collected and used for the isolation and screening of cellulose degradation bacteria using Luria Bertani (LB) Agar medium containing Carboxymethyl cellulose. The cellulase activity, biochemical characterization and phylogenetic analysis of the purified bacteria strains were characterized. A total of 3,833 OTUs were classified. The total microbial diversity detected in the female and male rats was 3,049 OTUs and 3,452 OTUs, respectively. The Shannon index revealed significant differences between the two groups (p < 0.05), though they were all captive and had the same feeding conditions. At the phylum level, Firmicutes, Bacteroidota, and Proteobacteria were prominent in the microbial community. At the genus level, the microbial community was dominated by Lachnospiraceae, Lactobacillus, Bacteroides, and Prevotella, but there was a significant difference between the two groups of bamboo rats; ~90 bacteria genus in the female group was significantly higher than the male group. Among them, Bacteroides, Colidextribacter, and Oscillibacter were significantly higher genera, and the genera of Lachnoclostridium, Oscillibacter, and Papillibacter had the highest FC value among the male and female bamboo rats. The KEGG function annotation and different pathways analysis revealed that membrane transport, carbohydrate metabolism, and amino acid metabolism were the most enriched metabolic pathways in the two groups, and multiple sugar transport system permease protein (K02025 and K02026), RNA polymerase sigma-70 factor (K03088), and ATP-binding cassette (K06147) were the three different KEGG pathways (p < 0.05). Two cellulose degradation bacteria strains-Bacillus subtilis and Enterococcus faecalis-were isolated and characterized from the feces of bamboo rats.

Antifungal Activity of Some Inorganic Salts on the Development of Fusarium spp., Causal Agents of Potato Tuber Dry Rot

Potato dry rot caused by Fusarium spp. is one of the main fungal diseases hampering the production and marketing of potato tubers in Cameroon. This study was carried out in order to improve the health status of potato tubers and their preservation. To achieve this objective, Fusarium spp. were isolated on the Potato Dextrose Agar (PDA) culture medium and identified on the basis of morphological and microscopic characteristics using reference documents. Antifungal activity of six inorganic salts was evaluated in vitro on the development of Fusarium spp. and on potato tubers artificially infected with these fungal species. The results showed that all the inorganic salts had a depressant effect on the development of Fusarium spp. Copper sulfate inhibited at 100%, the radial growth and sporulation of Fusarium spp. at all concentrations. Sodium metabisulfite and sodium carbonate inhibited at 100%, the growth and sporulation of all fungal species from a concentration of 6.4 mg/ml. No lesions were observed on inoculated potato tubers when they were treated with copper sulfate. These results suggest the implementation of a strategy to control dry rot of potato tubers through the use of inorganic salts, in order to reduce the losses caused by Fusarium spp.

Antibacterial Activities of Euphorbia hirta and Lantan camara Extracts on the Growth of Some Bacteria that Causes Banana Plant Disease

Bananas (Musa acuminata L.) are confronted with a number of challenges imposed by bacterial diseases which lead to huge yield losses. In order to improve the yields of the banana plantation, one of the recommended management strategies which would be inexpensive and environmentally friendly was developed. The objective of this study was to improve the sanitary state of banana plant. To achieve this objective, bacteria associated with the different banana organs were isolated on Nutrient Agar (culture medium) and their morphological identification was carried out based on the cultural characteristics and the colour of the bacterial walls observed under an optical microscope and using reference documents (bacteriological identification keys). Antibacterial activity of Lantana camara and Euphorbia hirta extracts were evaluated in vitro on agar medium aginst Xanthomonas campestris pv. Pseudomonas solanacearum and Ralstonia solanacearum. The results showed that the banana plant harbours a diversity of bacteria species, the most frequent being Ralstonia solanacearum as it was isolated from all infected organs. The yields of the equeous extacts of Euphorbia hirta and Lantana camara were particularly high (7% and 13%) respectively, compared to those of the ethanolic extracts which were lower. Aqueous extracts of Euphorbia hirta, at 50 mg/ml, had bactericidal activity against Ralstonia solanacearum and Pseudomonas celebensis. Aqueous extracts of Lantana camara, at concentrations of 25 mg/ml and 50 mg/ml, exhibited bactericidal activity against P. celebensis. Meanwhile, the ethanolic extracts of L. camara, at 50 mg/ml had bacteriostatic activity against the development of P. celebensis. Ethanolic extracts of E. hirta had bactericidal activity on the growth of Xanthomonas campestris at 25 mg/ml and 50 mg/ml. The same activity was obtained with P. celebensis at 50 mg/ml. These results suggest that the aqueous and ethanol extracts of the tested plants at high concentrations could be used as alternatives to chemical products in the fight against banana diseases especially Xanthomonas, Pseudomonas and Ralstonia. Hence further studies need to be undertaken in order to isolate the active compounds from these extracts with bactericidal potential.

Mushrooms Adapted to Seawater: Two New Species of Candolleomyces (Basidiomycota, Agaricales) from China.

Marine fungi have been studied for a long history in many realms, but there are few reports on marine mushrooms. In this study, marine fungi with conspicuous subglobose sequestrate basidioma were discovered from mangrove forests in South China. They grow on the deadwood of mangroves in the intertidal zone, periodically submerging into seawater due to the tide. Some marine animals were observed to nest in their basidiomata or consume them as food. The pileus-gleba-inner veil complex (PGI) of the basidioma was observed to be detached from the stipe and transferred into seawater by external forces, and drifting on sea to spread spores after maturity. The detachment mechanism of their PGIs was revealed through detailed microscopic observations. The contrast culturing experiment using freshwater and seawater potato dextrose agar media showed they have probably obligately adapted to the marine environment. Based on morphological and molecular phylogenetic evidence, two new species of Candolleomyces (Basidiomycota, Agaricales), namely C. brunneovagabundus and C. albovagabundus, were described. They are similar and close to each other, but can be distinguished by the size and color of the basidioma, and the size of the basidiospores.

Isolation and Molecular Identification of Proteolytic Bacteria from Rusip an Indonesian Fermented Food

Processing fish with the fermentation method can increase the nutritional content, bring out a distinctive taste, and make the fish last longer. Rusip is a type of complementary food from Bangka Belitung in the form of fish sauce that has been rotten or fermented. Rusip is made from fermented anchovy (Clupeoides borneensis). The concept of fermentation in Rusip is the breakdown of complex protein molecules by proteolytic bacteria into simple amino acid components.The purpose of this study was to determine the presence of proteolytic bacteria that play a role in rusip fermentation.The media used in screening proteolytic bacteria is skim milk agar (SMA) media. Then it was incubated at 37oCfor 24 hours. Bacterial colonies that form a clear zone on the media indicate that these bacteria can degrade protein. After the isolation process and isolates were obtained, the bacteria were characterized by Gram staining method and molecular identification.The results showed that there were proteolytic bacteria in Rusip food, and the 3 best isolates that produced protease enzymes were identified as Bacillus cereus.

Isolation and Identification of Fungal Species from the Insect Pest Callosobruchus maculatus (F.)

Cowpea seed beetle Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) is one of the most common and economically important pests of stored cereal products worldwide. Furthermore, these beetles can act as vectors for several fungal post-harvest diseases. The current research aimed to isolate the fungi associated with adults of C. maculatus (F.) (Coleoptera: Bruchidae). The present study aimed to isolate and identify fungi associated with adult insects and evaluate their potential as biocontrol agents against the cowpea beetle, C. maculatus. In this study, we collected C. maculatus adults from the Babylon local market and five regions (Hilah, Mahaweel, Al-Mashrou', Al-Haswa and Al-Musayyab) with no history of insecticide exposure. A potato dextrose agar medium was used to isolate the fungi attached to the surfaces of adult beetles. By Morphological and microscopic examination, Molecular identification and Sequencing analysis, Twenty-seven genera of fungi were isolated and identified from adult C. maculatus, including Aspergillus, Penicillium, Candida spp, white mycelium, Curvularia, Cladosporium, Chaetomium, Stachybotrys, Rhizopus, Drechslera, Scopulariopsis, Paecilomyces, Mucor, Geotrichum candidum, Streptomyces, Sarocladium, Beauveria bassiana, Talaromyces varians, Sporothrix flocculasa, Pseudozyma flocculasa, and Isaria fumosorosea. The molecular weights of the PCR products for the fungi isolates ranged from 650-700 bp, and the amplified ITS1-5.8S-ITS4 rDNA region of indigenous entomopathogenic fungal isolates showed a high sequence similarity (99-100%) with B. bassiana and M. anisopliae deposited in the NCBIGenebank. The phylogenetic tree analysis of the ITS region sequences showed a high degree of similarity between the isolates under study, ranging from 96.5-100. However, there were differences in the lines among clades, indicating genetic variation possibly due to mating or mutations in different environments. Our study suggests that the storage pest, C. maculatus, would be essential in spreading fungal contaminants and consequently increasing mycotoxin contamination in stored Cowpea. Keywords: Callosobruchus maculatus, Isolation, Fungi

Isolation and Identification of Fungal Species from the Insect Pest Callosobruchus maculatus (F.)

Cowpea seed beetle Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) is one of the most common and economically important pests of stored cereal products worldwide. Furthermore, these beetles can act as vectors for several fungal post-harvest diseases. The current research aimed to isolate the fungi associated with adults of C. maculatus (F.) (Coleoptera: Bruchidae); the present study aimed to isolate and identify fungi associated with adult insects and evaluate their potential as biocontrol agents against the cowpea beetle, C. maculatus. In this study, we collected C. maculatus adults from the Babylon local market and five regions (Hilah, Mahaweel, Al-Mashrou’, Al-Haswa and Al-Musayyab) with no history of insecticide exposure. A potato dextrose agar medium was used to isolate the fungi attached to the surfaces of adult beetles. By Morphological and microscopic examination, Molecular identification and Sequencing analysis, Twenty-seven genera of fungi were isolated and identified from adult C. maculatus, including Aspergillus, Penicillium, Candida spp, white mycelium, Curvularia, Cladosporium, Chaetomium, Stachybotrys, Rhizopus, Drechslera, Scopulariopsis, Paecilomyces, Mucor, Geotrichum candidum, Streptomyces, Sarocladium, Beauveria bassiana, Talaromyces varians, Sporothrix flocculasa, Pseudozyma flocculasa, and Isaria fumosorosea. The molecular weights of the PCR products for the fungi isolates ranged from 650-700 bp, and the amplified ITS1-5.8S-ITS4 rDNA region of indigenous entomopathogenic fungal isolates showed a high sequence similarity (99-100%) with B. bassiana and M. anisopliae deposited in the NCBIGenebank. The phylogenetic tree analysis of the ITS region sequences showed a high degree of similarity between the isolates under study, ranging from 96.5-100. However, there were differences in the sequences among clades, indicating genetic variation possibly due to mating or mutations in different environments. Our study suggests that the storage pest, C. maculatus, would be essential in spreading fungal contaminants and consequently increasing mycotoxin contamination in stored cowpeas. Keywords: Callosobruchus maculatus, Isolation, Fungi.

Draft genome sequences of Listeria monocytogenes strains of sequence type 1733 which constituted a pseudo-outbreak due to contaminated blood agar media.

We report genome sequences of Listeria monocytogenes sequence type (ST) 1733 from a 2013 pseudo-outbreak, where L. monocytogenes isolation from non-sterile sites (urine, wound, or abscess) was an artifact from contaminated sheep blood in the isolation media. Two ST1733 strains from wound and urine in 2005 are also reported.

Evaluation of substrate specificity and catalytic promiscuity of Bacillus albus cellulase: an insight into in silico proteomic study aiming at enhanced production of renewable energy

Cellulases are enzymes that aid in the hydrolysis of cellulosic fibers and have a wide range of industrial uses. In the present in silico study, sequence alignment between cellulases from different Bacillus species revealed that most of the residues are conserved in those aligned enzymes. Three dimensional structures of cellulase enzymes from 23 different Bacillus species have been predicted and based on the alignment between the modeled structures, those enzymes have been categorized into 7 different groups according to the homology in their conformational folds. There are two structural contents in Gr-I cellulase namely β1-α2 and β3-α5 loops which varies greatly according to their static position. Molecular docking study between the B. albus cellulase and its various cellulosic substrates including xylanoglucan oligosaccharides revealed that residues viz. Phe154, Tyr258, Tyr282, Tyr285, and Tyr376 of B. albus cellulase are significantly involved in formation stacking interaction during enzyme-substrate binding. Residue interaction network and binding energy analysis for the B. albus cellulase with different cellulosic substrates depicted the strong affinity of XylGlc3 substrate with the receptor enzyme. Molecular interaction and molecular dynamics simulation studies exhibited structural stability of enzyme-substrate complexes which are greatly influenced by the presence of catalytic promiscuity in their substrate binding sites. Screening of B. albus in carboxymethylcellulose (CMC) and xylan supplemented agar media revealed the capability of the bacterium in degrading both cellulose and xylan. Overall, the study demonstrated B. albus cellulase as an effective biocatalyst candidate with the potential role of catalytic promiscuity for possible applications in biofuel industries. Communicated by Ramaswamy H. Sarma

Viability of Bacillus coagulans spores with ofloxacin &amp; ornidazole: Implications for gut health

The balance of gut microbiota significantly impacts host health. Disruption of the natural gut flora, often caused by infections or the use of broad-spectrum antimicrobial drugs, can lead to dysbiosis, causing gastrointestinal disorders, such as diarrhea. Probiotics show promising outcomes in restoring gut health, but concerns remain about their interaction with antimicrobials and the viability of spores in the intended gut location. To address these uncertainties, the current study was devised to evaluate how well spores-forming bacteria endure and develop in an environment where antimicrobial agents are present.: The study investigated the survival and growth of spores under the influence of broad-spectrum antimicrobial agents, Ofloxacin and Ornidazole. To cultivate the spores in the presence of these antimicrobials, a mixture of MRS broth and PNY agar media was used. The number of colonies that developed were measured to assess the extent of spore survival and germination.In a simulated environment resembling human intestinal pH, spores exhibited viability. Starting with an initial count of 1.38 billion CFU, the spores multiplied to 8.75 billion CFU at 24 hours and further reached to 86.25 billion CFU at 72 hours in the presence of Ofloxacin and Ornidazole. On the other hand, the viable count reached to the level of 88 billion CFU in the absence of antimicrobial agents.This study offers evidence that spores are able to remain viable and germinate when co-administered with Ofloxacin and Ornidazole.