Tracing probiotic producing bacterial species from gut of buffalo (Bubalus bubalis), South-East-Asia / Rastreando espécies bacterianas produtoras de probióticos de intestino de búfalo (Bubalus bubalis), sudeste da Ásia

Due to extensive application of antibiotics as growth promoters in animal feed, antimicrobial resistance has been increased. To overcome this challenge, rumen microbiologists search for new probiotics to improve the rate of livestock production. The present study was aimed to isolate and evaluate breed-specific lactic acid bacteria (LAB) as potential animal probiotics. The current study was conducted during 10 months from July 2020 to April 2021, in which a total of n=12 strains were isolated from different samples including milk, rumen, and feces of Nilli Ravi Buffaloes. These isolates were evaluated for their antimicrobial potential against common animal pathogens (Bacillus spp., E. coli, Staphylococcus aureus, Salmonella spp., Listeria spp.). All the isolates were identified using 16S rRNA gene sequencing and the phylogenetic analyses inferred that these strains showed close relations to the species of various genera; Enterococcus lactis, Pediococcus pentosaceus, Bacillus subtilis Weissella cibaria, Weissella soli, Bacillus tequilensis, Weissella bombi, Bacillus licheniformis, Lactococcus lactis, Bacillus megaterium, Lactobacillus ruminis, and Lactococcus lactis. NMCC-Ru2 has exhibited the enormous potential of antimicrobial activity, 28 mm, for Salmonella typhimurium;23 mm for Listeria monocytogenes 21 mm for E.coil. Highest resistance was seen in NMCC-Ru2 agasint test antbiotic, like 25.5 mm for Tetracycline. Overall results revesl that the probiotic profile of isolates was achieved using standard criteria, particularly with animal probiotic properties

Devido à extensa aplicação de antibióticos como promotores de crescimento na alimentação animal, a resistência aos antimicrobianos aumentou. Para superar esse desafio, os microbiologistas do rúmen buscam novos probióticos para melhorar a produtividade do gado. O presente estudo teve como objetivo isolar e avaliar bactérias lácticas específicas de raças (BAL) como potenciais probióticos animais. 12 cepas foram isoladas de diferentes amostras, incluindo leite, rúmen e fezes de búfalos Nilli Ravi. Esses isolados foram avaliados quanto ao seu potencial antimicrobiano contra patógenos animais comuns (Bacillus spp., E. coli, Staphylococcus aureus, Salmonella spp., Listeria spp.). Todos os isolados foram identificados por meio do sequenciamento do gene 16S rRNA e as análises filogenéticas inferiram que essas cepas apresentaram estreita relação com as espécies de vários gêneros; Enterococcus lactis, Pediococcus pentosaceus, Bacillus subtilis, Weissella cibaria, Weissella soli, Bacillus tequilensis, Weissella bombi, Bacillus licheniformis, Lactococcus lactis, Bacillus megaterium, Lactobacillus ruminis e Lactococcus lactis. O perfil probiótico dos isolados foi obtido usando critérios padrão, particularmente com propriedades probióticas animais.

Tracing probiotic producing bacterial species from gut of buffalo (Bubalus bubalis), South-East-Asia

Abstract Due to extensive application of antibiotics as growth promoters in animal feed, antimicrobial resistance has been increased. To overcome this challenge, rumen microbiologists search for new probiotics to improve the rate of livestock production. The present study was aimed to isolate and evaluate breed-specific lactic acid bacteria (LAB) as potential animal probiotics. The current study was conducted during 10 months from July 2020 to April 2021, in which a total of n=12 strains were isolated from different samples including milk, rumen, and feces of Nilli Ravi Buffaloes. These isolates were evaluated for their antimicrobial potential against common animal pathogens (Bacillus spp., E. coli, Staphylococcus aureus, Salmonella spp., Listeria spp.). All the isolates were identified using 16S rRNA gene sequencing and the phylogenetic analyses inferred that these strains showed close relations to the species of various genera; Enterococcus lactis, Pediococcus pentosaceus, Bacillus subtilis Weissella cibaria, Weissella soli, Bacillus tequilensis, Weissella bombi, Bacillus licheniformis, Lactococcus lactis, Bacillus megaterium, Lactobacillus ruminis, and Lactococcus lactis. NMCC-Ru2 has exhibited the enormous potential of antimicrobial activity, 28 mm, for Salmonella typhimurium;23 mm for Listeria monocytogenes 21 mm for E.coil. Highest resistance was seen in NMCC-Ru2 agasint test antbiotic, like 25.5 mm for Tetracycline. Overall results revesl that the probiotic profile of isolates was achieved using standard criteria, particularly with animal probiotic properties

Resumo Devido à extensa aplicação de antibióticos como promotores de crescimento na alimentação animal, a resistência aos antimicrobianos aumentou. Para superar esse desafio, os microbiologistas do rúmen buscam novos probióticos para melhorar a produtividade do gado. O presente estudo teve como objetivo isolar e avaliar bactérias lácticas específicas de raças (BAL) como potenciais probióticos animais. 12 cepas foram isoladas de diferentes amostras, incluindo leite, rúmen e fezes de búfalos Nilli Ravi. Esses isolados foram avaliados quanto ao seu potencial antimicrobiano contra patógenos animais comuns (Bacillus spp., E. coli, Staphylococcus aureus, Salmonella spp., Listeria spp.). Todos os isolados foram identificados por meio do sequenciamento do gene 16S rRNA e as análises filogenéticas inferiram que essas cepas apresentaram estreita relação com as espécies de vários gêneros; Enterococcus lactis, Pediococcus pentosaceus, Bacillus subtilis, Weissella cibaria, Weissella soli, Bacillus tequilensis, Weissella bombi, Bacillus licheniformis, Lactococcus lactis, Bacillus megaterium, Lactobacillus ruminis e Lactococcus lactis. O perfil probiótico dos isolados foi obtido usando critérios padrão, particularmente com propriedades probióticas animais.

Antibacterial Activity of Economically Important Medicinal Plants in Pakistan Against Different Bacterial Strains.

The emergence of medication resistance and unfavorable side effects from existing antibiotics has prompted the quest for novel antimicrobial agents over the last 2 decades. Plant extracts have been shown to have antibacterial effects in numerous studies. The objective of this study was the evaluation of the antibacterial effect of economically important medicinal plants found in Pakistan. Onosma bracteatum (flowers and leaves), Viola odorata (flowers and leaves), Cuscuta reflexa (whole plant), Swertia chirata (whole plant), and Fagonia arabica (whole plant) were used against Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa. Water and ethanol extracts were obtained from different parts of the plants. To evaluate the antibacterial effect of these plants, qualitative assay agar well diffusion method was performed. The minimum inhibitory concentration (MIC) was determined by the broth micro dilution method. Results revealed that the highest inhibition zone (18 mm) was shown by ethanol extract of V odorata flower against P aeruginosa. Ethanol extract of C reflexa plants is best for all 3 tested microbes (P aeruginosa, B subtilis, and E coli). The results concluded that all these plants have abilities to fight against these tested bacteria. Ethanol extract of V odorata flower has the highest activity against P aeruginosa.

Evaluation of Phytochemical Contents, Antimicrobial, and antioxidant Potential of Haloxylon Griffithii Collected From Northern Region of Balochistan, Pakistan.

Background: The therapeutic potential of Haloxylon griffithii found in northern region of Balochistan, so far has been neglected. Purpose: The current study was aimed to assess the phytochemicals and pharmacological potential of fractions isolated from H griffithii. Research Design: During phytochemicals analysis of H griffithii using GC/MS showed various bioactive compounds like alkaloids, flavonoids, terpenoids, tannins, saponins, and carboxylic acids. In vitro antioxidant activity of H griffithii was determined by 2, 2'- diphenyl-1-picrylhydrazyl (DPPH) assay. Disc diffusion method was used to evaluate the antimicrobial activity. Results: The quantitative analysis of ethyl acetate showed highest total flavonoid contents (1.19 ± .05) while ethanol with lowest value (.52 ± .01). The total phenolic contents in ethyle acetate was 1.50 ± .42, whereas ethanol showed lowest value (.77 ± .02). Ethanol exhibited excellent (88.68 ± 3.0) free radical scavenging potential measured by 1,1-diphenyl-2-picryl-hydrazyl radical scavenging assay. For antimicrobial activity, different bacterial and fungal strains like B subtilis, S aureus, E coli, S typhi, C albicans, and A. niger were selected. The essential oil showed maximum % inhibition diameter (9 mm) against B. Subtillus and (5 mm) against C albicans, respectively. The ethyl acetate presented % inhibition diameter (9 mm) against S aureus and (6 mm) against A niger. Anti-urease activity also showed positive response. Conclusions: The presence of high (%) bioactive compounds with great therapeutic potential suggest that H griffithii can be used as natural alternative of synthetic drugs without side effects.

Efficacy of Saccharothrix algeriensis NRRL B-24137 to suppress Fusarium oxysporum f.sp. vasinfectum induced wilt disease in cotton.

Fusarium cotton wilt is a devastating disease of the cotton crop throughout the world, caused by Fusarium oxysporum f.sp. vasinfectum (FOV). Chemical control has many side effects, so, biological controls have been widely used for the management of Fusarium wilt. This study aimed to investigate the possible use of an actinomycetes Saccharothrix algeriensis (SA) NRRL B-24137 to control FOV. To access in-vitro anti-Fusarium ability of SA NRRL B-24137, dual culture assay, spore germination and seed germination tests were carried out. Following in-vitro investigations, several pot tests in a greenhouse environment were used to evaluate the biological control potential of SA NRRL B-24137 against FOV. Dual culture assay and spore germination revealed that SA NRRL B-24137 showed significant anti-Fusarium activity.During spore germination 87.77% inhibition of spore germination were observed. In pot experiments, SA NRRL B-24137 primed cotton seeds resulted in a 74.0% reduction in disease incidence. In soil there was a significant reduction in FOV spores in the presence of SA NRRL B-24137. Positive correlation was also observed on different concentrations of SA NRRL B-24137 towards FOV reduction. The results of this study showed that SA NRRL B-24137 has the potential to be employed as a biocontrol agent against Fusarium cotton wilt, improving cotton growth characteristics and yield.

Biosorptive Potential of Pseudomonas species RY12 Toward Zinc Heavy Metal in Agriculture Soil Irrigated with Contaminated Waste Water.

Industrial waste is mainly responsible for accumulating Zn (II) in the soil, which needs to be removed to avoid its bioaccumulation and hazardous effects on the environment. In a recent study, the potential of the RY12 strain was evaluated as a biosorbent of Zn (II) ions in an aqueous medium. Different microbiological techniques like biochemical, molecular characterization, and 16S rRNA gene sequencing were used for the identification of RY12. The impact of different parameters such as the initial zinc ion concentration, pH, temperature, and the removal of other metals such as manganese, lead, cobalt, silver, copper, mercury, and chromium was also evaluated on the reduction of Zn (II). Fourier Transform Infrared spectroscopy (FTIR) was also carried out to investigate the role of cellular surfaces in the sorption of Zn+2 ions. Both biochemical and phylogenetic analyses established that strain RY12 Pseudomonas sp. capable of reducing Zn+2 up to 89% at 28°C (pH = 6.5; initial Zn+2 concentration = 200 mg/L). The FTIR analysis revealed that the bacterial cell wall's amino, carboxyl, and phosphate groups were involved in the reaction with Zn (II). Our findings suggest that Pseudomonas sp. RY12 is a proficient bacterium for removing zinc from industrial waste and could be a valuable bioremediation agent.

Green Synthesis of Silver Nanoparticles (AgNPs), Structural Characterization, and their Antibacterial Potential.

Background: In the field of nanotechnology, the metallic nanoparticles are of remarkable interest because of their unique electronic, magnetic, chemical, and mechanical properties. Purpose: In the present work, silver nanoparticles (AgNPs) were synthesized using bio-reduction method. Research Design: Silver nitrate was used as metallic precursor and the extract of Moringa oleifera leaves with different concentrations was used as reducing as well capping agent. The extract exhibited strong potential in rapid reduction of silver ions for the synthesis of silver nanoparticles. The synthesized silver nanoparticles were characterized by UV-visible spectroscopy, X-ray diffraction (XRD), and scanning electron microscopy (SEM) techniques. Results: The absorption SPR peaks appeared in the range of 415 to 439 nm. SEM analysis exhibited that particles were spherical in shape with size distribution range from 10 nm to 25 nm. The synthesized silver nanoparticles were pure crystalline in nature as confirmed by the XRD spectra with average crystallite size 7 nm. In vitro antibacterial activity of the prepared silver nanoparticles colloidal samples as well the extract was studied using different concentrations of AgNPs (C1 = 100 µg/ml, C2 = 50 µg/ml, C3 = 25 µg/ml) by well diffusion method against Gram negative Escherichia coli. The antibacterial performance was assessed by measuring the zone of inhibition (ZOI). Conclusions: The results suggested that AgNPs prepared by green approach can be considered as an alternative antibacterial agent.

Tracing probiotic producing bacterial species from gut of buffalo (Bubalus bubalis), South-East-Asia.

Due to extensive application of antibiotics as growth promoters in animal feed, antimicrobial resistance has been increased. To overcome this challenge, rumen microbiologists search for new probiotics to improve the rate of livestock production. The present study was aimed to isolate and evaluate breed-specific lactic acid bacteria (LAB) as potential animal probiotics. The current study was conducted during 10 months from July 2020 to April 2021, in which a total of n=12 strains were isolated from different samples including milk, rumen, and feces of Nilli Ravi Buffaloes. These isolates were evaluated for their antimicrobial potential against common animal pathogens (Bacillus spp., E. coli, Staphylococcus aureus, Salmonella spp., Listeria spp.). All the isolates were identified using 16S rRNA gene sequencing and the phylogenetic analyses inferred that these strains showed close relations to the species of various genera; Enterococcus lactis, Pediococcus pentosaceus, Bacillus subtilis Weissella cibaria, Weissella soli, Bacillus tequilensis, Weissella bombi, Bacillus licheniformis, Lactococcus lactis, Bacillus megaterium, Lactobacillus ruminis, and Lactococcus lactis. NMCC-Ru2 has exhibited the enormous potential of antimicrobial activity, 28 mm, for Salmonella typhimurium;23 mm for Listeria monocytogenes 21 mm for E.coil. Highest resistance was seen in NMCC-Ru2 agasint test antbiotic, like 25.5 mm for Tetracycline. Overall results revesl that the probiotic profile of isolates was achieved using standard criteria, particularly with animal probiotic properties.

Breast Cancer Risk and Human Papillomavirus Infection: A Bradford Hill Criteria Based Evaluation.

BACKGROUND: The association between human papillomavirus (HPV) and human breast cancer (BC) has already been thoroughly studied worldwide with contradictory findings. Although the researchers have tried to minimize the conflict using statistical meta-analysis because of its shortcomings, there is still a need to evaluate the correlation between HPV and BC using any additional method. OBJECTIVES: This study was launched to investigate the correlation between HPV and BC through the application of Bradford Hill criteria postulates. METHODS: Population-wide studies associating HPV with BC were searched using the PubMed database. Then, the information of HPV burden in BC, normal/benign samples was analyzed, and ultimately Bradford Hill criteria postulates were applied on the collected evidence to explore the relationship between HPV and BC. In addition, to make the outcomes more authentic, we also reviewed the methodologies of previous studies to address the propensity of false results. RESULTS: After a careful evaluation of the obtained data against major Bradford Hill criteria postulates, it was noted that all these postulates, including strength, consistency, biological gradient, temporality, plausibility, experiment, specificity, and analogy were not fulfilled. CONCLUSION: The results of the present study have failed to establish a causal association between HPV and BC, but they did suggest HPV as a cause-effective agent or at least a co-participant in the pathogenesis of BC. Because of the weakness of association, particularly the lack of consistency between studies and the lack of effect specificity, more research into Bradford Hill criteria postulates is required.

Integrative analysis reveals methylenetetrahydrofolate dehydrogenase 1-like as an independent shared diagnostic and prognostic biomarker in five different human cancers.

BACKGROUND: Defects in methylenetetrahydrofolate dehydrogenase 1-like (MTHFD1L) expression have earlier been examined in only a few human cancers. OBJECTIVES: Multi-omics profiling of MTHFD1L as a shared biomarker in distinct subtypes of human cancers. METHODS: In the current study, for the multi-omics analysis of MTHFD1L in 24 major subtypes of human cancers, a comprehensive in silico approach was adopted to mine different open access online databases including UALCAN, Kaplan-Meier (KM) plotter, LOGpc, GEPIA, Human Protein Atlas (HPA), Gene Expression across Normal and Tumor tissue (GENT2), MEXPRESS, cBioportal, STRING, DAVID, TIMER, and Comparative Toxicogenomics Database (CTD). RESULTS: We noticed that the expression of MTHFD1L was significantly higher in all the analyzed 24 subtypes of human cancers as compared with the normal controls. Moreover, MTHDF1L overexpression was also found to be significantly associated with the reduced overall survival (OS) duration of Bladder urothelial cancer (BLCA), Head and neck cancer (HNSC), Kidney renal papillary cell carcinoma (KIRP), Lung adenocarcinoma (LUAD), and Uterine corpus endometrial carcinoma (UCEC). This implies that MTHFD1L plays a significant role in the development and progression of these cancers. We further noticed that MTHFD1L was also overexpressed in BLCA, HNSC, KIRP, LUAD, and UCEC patients of different clinicopathological features. Pathways enrichment analysis revealed the involvement of MTHFD1L-associated genes in five diverse pathways. We also explored few interesting correlations between MTHFD1L expression and its promoter methylation, genetic alterations, CNVs, and between CD8+ T immune cells level. CONCLUSION: In conclusion, our results elucidated that MTHFD1L can serve as a shared diagnostic and prognostic biomarker in BLCA, HNSC, KIRP, LUAD, and UCEC patients of different clinicopathological features.

Multi-Omics Analysis Identified TMED2 as a Shared Potential Biomarker in Six Subtypes of Human Cancer.

INTRODUCTION: Cancer is one of the most common malignancies and the leading cause of death worldwide. As a member of the transmembrane emp24 domain (Tmed)/p24 family of proteins, TMED2 expression variations have been documented earlier in only a few subtypes of human cancers, and the multi-omics profiling of TMED2 as a shared biomarker in different other subtypes of human cancers remains to be uncovered. METHODS: In the current study, TMED2 multi-omics analysis in 24 major subtypes of human cancer was performed using different authentic online databases and bioinformatics analysis including UALCAN, Kaplan-Meier (KM) plotter, Human Protein Atlas (HPA), GENT2, MEXPRESS, cBioportal, STRING, DAVID, TIMER, and CTD. RESULTS: In general, the TMED2 expression in 24 major subtypes of human cancers was higher relative to normal controls and was also strongly associated with the lower overall survival (OS) and relapse-free survival (RFS) duration of CESC, ESCA, HNSC, KIRC, LIHC, and LUAD patients. This implies that TMED2 plays a significant role in the development and progression of these cancers. Furthermore, the TMED2 overexpression was also correlated with different clinicopathological features of CESC, ESCA, HNSC, KIRC, LIHC, and LUAD patients. TMED2-associated genes network was involved in 3 diverse pathways, and finally, few stronger correlations were also explored between TMED2 expression and its promoter methylation level, genetic alterations, and CD8+ T immune cells level. CONCLUSION: In conclusion, via this in silico study, we have elucidated that TMED2 can serve as a shared diagnostic and prognostic biomarker in CESC, ESCA, HNSC, KIRC, LIHC, and LUAD patients of different clinicopathological features but, further in vitro and in vivo research should be carried out to confirm these findings.

CTHRC1 expression is a novel shared diagnostic and prognostic biomarker of survival in six different human cancer subtypes.

According to the previous reports, the collagen triple helix repeat containing 1 (CTHRC1) causes tumorigenesis by modulating the tumor microenvironment, however, the evidence is limited to a few human cancer subtypes. In the current study, we analyzed and validated the CTHRC1 expression variations in 24 different human cancer tissues paired with normal tissues using publically available databases. We observed that CTHRC1 was overexpressed in all the 24 major subtypes of human cancers and its overexpression was significantly associated with the reduced overall survival (OS) duration of head and neck squamous cell carcinoma (HNSC), kidney renal clear cell carcinoma (KIRC), liver hepatocellular carcinoma (LIHC), Lung adenocarcinoma (LUAD), stomach adenocarcinoma (STAD), and Uterine corpus endometrial carcinoma (UCEC). This implies that CTHRC1 plays a significant role in the development and progression of these cancers. We further noticed that CTHRC1 was also overexpressed in HNSC, KIRC, LIHC, LUAD, STAD, and UCEC patients of different clinicopathological features. Pathways enrichment analysis revealed the involvement of CTHRC1 associated genes in seven diverse pathways. We also explored few interesting correlations between CTHRC1 expression and promoter methylation, genetic alterations, CNVs, CD8+ T immune cells infiltration, and tumor purity. In conclusion, CTHRC1 can serve as a shared diagnostic and prognostic biomarker in HNSC, KIRC, LIHC, LUAD, STAD, and UCEC patients of different clinicopathological features.

Saccharothrix Algeriensis NRRL B-24137 Potentiates Chemical Fungicide Carbendazim in Treating Fusarium Oxysporum f.sp. Vasinfectum-Induced Cotton Wilt Disease.

Cotton (Gossypium hirsutum) wilt is one of the destructive disease caused by Fusarium oxysporum f. sp. vasinfectum and lead to 100% yield loss under favorable conditions. This study aims to estimate the potential of biological control agents Saccharothrix algeriensis NRRL B-24137 (SA) and chemical fungicides against cotton wilt pathogen under in-vitro and in-vivo conditions. The in-vitro study revealed that carbendazim showed maximum mycelia growth inhibition with a mean of 91% over control, which was further validated in glasshouse assay. In-vitro dual culture test of biocontrol agents with F. oxysporum determined that SA had a potential to inhibit mycelia growth by 68% compared to control. Further in glasshouse assay, the combination of the SA and carbendazim (10 µg/mL) showed a significant (p < 0.05) disease control. Moreover, results demonstrated that carbendazim and SA remarkably decreased the disease development up to 83% and subsequently, significant improvement was observed in the plant growth parameters (plant length, root length, and plant weight) compared to untreated plants. Conclusively, exploration and utilization of bioagent for fungal diseases in cotton may provide a better line with maximum efficacy and with lesser adverse effects, which will pave a way toward better consequences in fungal treatments.

In vivo antiviral potential of Glycyrrhiza glabra extract against Newcastle disease virus.

Newcastle disease is highly infectious viral disease causing huge economic losses worldwide. These losses can be prevented by control of viral diseases. Medicinal plants have been traditionally used for treatment of different diseases since long. In this study the effect of extracts from Glycyrrhiza glabra leaves are investigated against Newcastle disease virus (NDV) by an in-vivo assay. Seven groups of nine-day-old embryonated chicken eggs were inoculated with various treatments of different plant extracts. All the groups except uninoculated negative control group were inoculated with velogenic NDV strain; five groups received different concentrations of the three extracts. Daily observe the rate of embryo survival. Allantoic fluid from treated eggs was collected for hem agglutination test. Results showed that embryo survival rate was higher 300µg/mL treated group as all the extracts showed antiviral activity. Similarly, the plant extracts effectively control virus as no viruses were identified in the allantoic fluids of all groups treated with low doses of plant. The current results have clearly verified that all the extracts especially that of methanol 300µg/mL from leaves of Glycyrrhiza glabra have strong antiviral activity against NDV in vivo.