Phenotypic and molecular characterization of clinically isolated antibiotics-resistant S. aureus (MRSA), E. coli (ESBL) and Acinetobacter 1379 bacterial strains.

Antibiotic-resistant bacteria causing nosocomial infections pose a significant global health concern. This study focused on examining the lipid profiles of both non-resistant and clinically resistant strains of Staphylococcus aureus (MRSA 1418), E. coli (ESBL 1384), and Acinetobacter 1379. The main aim was to investigate the relationship between lipid profiles, hydrophobicity, and antibiotic resistance so as to identify the pathogenic potential and resistance factors of strains isolated from patients with sepsis and urinary tract infections (UTIs). The research included various tests, such as antimicrobial susceptibility assays following CLSI guidelines, biochemical tests, biofilm assays, and hydrophobicity assays. Additionally, gas chromatography mass spectrometry (GC-MS) and GC-Flame Ionization Detector (GC-FID) analysis were used for lipid profiling and composition. The clinically isolated resistant strains (MRSA-1418, ESBL-1384, and Acinetobacter 1379) demonstrated resistance phenotypes of 81.80%, 27.6%, and 63.6%, respectively, with a multiple antibiotic resistance index of 0.81, 0.27, and 0.63. Notably, the MRSA-1418 strain, which exhibited resistance, showed significantly higher levels of hemolysin, cell surface hydrophobicity, biofilm index, and a self-aggregative phenotype compared to the non-resistant strains. Gene expression analysis using quantitative real-time PCR (qPCR). Indicated elevated expression levels of intercellular adhesion biofilm-related genes (icaA, icaC, and icaD) in MRSA-1418 (pgaA, pgaC, and pgaB) and Acinetobacter 1379 after 24 h compared to non-resistant strains. Scanning electron microscopy (SEM) was employed for structural investigation. These findings provide valuable insights into the role of biofilms in antibiotic resistance and suggest potential target pathways for combating antibiotic-resistant bacteria.

Comparative Analysis of Various Materials Used for Mastoid Cavity Obliteration in Canal Wall Down Mastoid Surgery.

To study the surgical result and efficacy of different methods of mastoid obliteration with cavity care. This prospective study included 60 patients who had cholesteatoma, conducted in the Department of Ear Nose and Throat (ENT), Institute of Medical Sciences, Banaras Hindu University, Varanasi from July 2015 to July 2017. The mastoid cavity was obliterated with either muscle flap/bone dust/hydroxyapatite. detailed history otoscopic examination was done pre-operatively and follow up were recorded at 1 month and 3 months in postoperative period. 60 patients were included, who underwent canal wall down mastoid surgery. each group muscle flap (group 1), bone dust (group 2) and hydroxyapatite (group 3) included 20 patients, age group 31-40 year with its maximum incidence of 43.3%, Preop PTA value were almost equal in all group but on comparison at 1 month in postoperative period significant improvement was present in group 1 versus 2(0.021) and group 2 versus 3(0.003) but not in group 1 versus 3. Although at 3 month there were significant improvement was present in all groups. The incidence of pain, discharge, giddiness and wax formation were markedly reduced and healing of cavities was early and better in obliterated cavities done by muscle flap and bone dust material as compared to hydroxyapatite cavities, at the end of 3 months. outcome and quality of life was better and almost equal in muscle flap and bone dust material group as compared to hydroxyapatite group.

Draft genome sequence of methicillin-resistant Staphylococcus aureus strain D1418m22 isolated from human wound pus.

This study reveals the complete genome sequence of methicillin-resistant Staphylococcus aureus (MRSA) strain d1418m22, sourced from a Karnal, Haryana, human skin wound. Classified as community-associated MRSA, it features a 2.78-MB genome harboring Staphylococcus-specific genetic elements, encompassing 2,625 protein-coding genes and 18 antimicrobial resistance genes.

"In vitro antimicrobial and synergistic effect of fermented Indian zebu (Sahiwal) cow colostrum whey derived peptides with Lactobacillus rhamnosus against pathogenic bacteria".

Bioactive peptides (BAPs) have been found to promote health through various mechanisms. Among them, antimicrobial peptides are gaining recognition as promising novel treatments. This study aims to generate BAPs from bovine colostrum whey using the proteolytic activity of Lactobacillus rhamnosus C25 and to evaluate their potential antibacterial efficacy, including their ability to synergistic efficacy against resistant bacteria. Bioactive peptides were successfully generated from lactobacillus culture proteases that were cultivated through batch fermentation. The resulting peptide fractions were then evaluated for their antibacterial efficacy against a selection of strains, including E. coli ATCC25922, S. aureus MTCC1144, Acinetobacter baumannii ATCC 17978, as well as clinically isolated resistant strains of E. coli (ESBL 1384), Acinetobacter 1379, and S. aureus (MRSA 1418). Notably, the peptide fractions with a molecular weight of < 10 kDa (0-10 kDa) significantly increased the membrane permeability of both E. coli (70.30 ± 0.41%) and S. aureus (63.04 ± 0.31%) as assessed by the crystal violet assay. The checkerboard method was utilized to perform synergistic tests with peptides and antibiotics. The peptide fractions with a molecular weight of (< 10 kDa) demonstrated synergistic effects with several antibiotics, including gentamycin, Rifampicin, Levofloxacin, Ciprofloxacin, and Chloramphenicol, against the resistant ESBL 1384 strain, as indicated by ΣFICI values of 0.55, 0.53, 0.52, 0.54, and 0.52, respectively. Furthermore, the HT-29 cell line remained completely unaffected by both peptide fractions. These findings suggest that the < 10 kDa peptide fraction possesses significant antibacterial efficacy against both reference and ESBL 1384 resistant bacterial strain. Additionally, both MRSA 1418 and Acinetobacter 1379 displayed resistance to all fractions tested. To summarize the findings of this study, colostrum whey peptides with a broad spectrum of antimicrobial activity can be efficiently produced through fermentation. This method could prove valuable for both the pharmaceutical and food industries. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-023-05776-2.

Encapsulation and Assessment of Antidiabetic Potential of α-Lactalbumin-Derived Hydrolysates.

Dipeptidyl peptidase-IV (DPP-IV) is an exopeptidase mainly present in epithelial tissues of the liver, kidney, and intestine. It is involved in the cleavage of a variety of substrates including the incretin hormones like glucagon-like peptide-1 (GLP-1). GLP-1 binds to the GLP-1 receptors of pancreatic ß-cells and leads to ß-cell proliferation and increases insulin secretion through associated gene expression. In diabetes, a constant increase in the glucose level leads to glucotoxicity, which destroys pancreatic ß-cells, decreases the insulin level, and further increases the blood glucose level. Inhibition of DPP-IV is one of the strategies for the treatment of type 2 diabetes. In recent years, peptides derived from a variety of dietary proteins have been reported to exhibit inhibitory activity against the DPP-IV enzyme. Such peptides should also be protected from the action of digestive enzymes to keep their bioactivity intact. Therefore, the present investigation was aimed to evaluate the in vitro DPP-IV inhibition potential and in vivo antidiabetic potential of α-lactalbumin in non-encapsulated hydrolysate (NEH), freeze-dried encapsulated hydrolysate (FDEH), and emulsified encapsulated hydrolysate (EEH) forms. Percent DPP-IV inhibition by the NEH, FDEH, and EEH after simulated gastrointestinal digestion was 36 ± 2.28, 54 ± 2.02, and 64 ± 2.02, respectively. The oral administration of the NEH, FDEH, and EEH at a dose of 300 mg/kg body weight was evaluated in nicotinamide-streptozotocin-induced type 2 diabetic experimental rats in a study of 30 days. Rats in the diabetic control group showed an increase in the blood glucose level and liver function enzymes and a decrease in GLP-1, insulin, and antioxidative enzymes. Administration of hydrolysates reversed the parameters by lowering the blood glucose level and increasing GLP-1 and insulin levels in plasma. The blood lipid profile, liver enzyme (ALT, AST, and AP) levels, and catalase and superoxide dismutase activity were also found to be normalized and better managed in experimental diabetic rats.

Peptidomics-based identification of an antimicrobial peptide derived from goat milk fermented by Lactobacillus rhamnosus (C25).

Antimicrobial peptides (AMPs) are emerging as promising novel drug applicants. In the present study, goat milk was fermented using Lactobacillus rhamnosus C25 to generate bioactive peptides (BAPs). The peptide fractions generated were separated using ultrafiltration membranes with molecular weight cut-offs of 3, 5, and 10 kDa, and their antimicrobial activity toward Gram-positive and Gram-negative bacteria was investigated. Isolated AMPs were characterized using RP-HPLC and identified by LC-MS/MS. A total of 569 sequences of peptides were identified by mass spectrometry. Out of the 569, 36 were predicted as AMPs, 21 were predicted as cationic, and out of 21, 6 AMPs were helical peptides. In silico analysis indicated that the majority of peptides were antimicrobial and cationic in nature, an important factor for peptide interaction with the negative charge membrane of bacteria. The results showed that the peptides of <5 kDa exhibited maximum antibacterial activity against E. faecalis, E. coli, and S. typhi. Further, molecular docking was used to evaluate the potent MurD ligase inhibitors. On the basis of ligand binding energy, six predicted AMPs were selected and then analyzed by AutoDock tools. Among the six AMPs, peptides IGHFKLIFSLLRV (-7.5 kcal/mol) and KSFCPAPVAPPPPT (-7.6 kcal/mol), were predicted as a high-potent antimicrobial. Based on these findings, in silico investigations reveal that proteins of goat milk are a potential source of AMPs. This is for the first time that the antimicrobial peptides produced by Lactobacillus rhamnosus (C25) fermentation of goat milk have been identified via LC-MS/MS and predicted as AMPs, cationic charges, helical structure in nature, and potent MurD ligase inhibitors. These peptides can be synthesized and improved for use as antimicrobial agents. PRACTICAL APPLICATIONS: Goat milk is considered a high-quality source of milk protein. According to this study, goat milk protein is a potential source of AMPs, Fermentation can yield goat milk-derived peptides with a broad antibacterial activity spectrum at a low cost. The approach described here could be beneficial in that the significant AMPs can be synthesized and used in the pharmaceutical and food industries.

Antimicrobial bioactive peptides from goat Milk proteins: In silico prediction and analysis.

The main goal of this study was to assess the potential proteins of goat milk (i.e. α-s1-casein, α-s2-casein, ß-casein, κ-casein, α-lactoglobulin and ß-lactalbumin) as precursors of antimicrobial peptides (AMPs). Bioinformatics tools such as BIOPEP-UWM (enzyme action) were used for the in silico gastrointestinal digestion via a cocktail of pepsin, trypsin, and chymotrypsin A. The antimicrobial activity of peptides was predicted by using four algorithms, including Random Forest, Support Vector Machines, Artificial Neural Network and Discriminant Analysis on CAMPR3 online server, which works on Hidden Markov Models. Different online tools predicted the physiochemical properties, allergenicity, and toxicity of peptides as well. In silico gastrointestinal digestion simulation of proteins by enzymes cocktail yielded a total of 83 potential AMPs, with thirteen peptides being confident by all four algorithms. More AMPs were released from ß-casein (21) than from ß-lactoglobulin (16), α-s1-casein (15), α-s2-casein (12), κ-casein (11) and α-lactalbumin (9). A total of 17 peptides were cationic, and the majority of the peptides were extended AMPs. These peptides were released from α-s1-casein (SGK, IQK), α-s2-casein (SIR, AIH, TQPK), ß-casein (GPVR, AVPQR, AIAR, GVPK, SQPK, PVPQK, IH, VPK), k-casein (AIPPK, QQR, IAK, TVPAK). All of the AMPs were anticipated to be non-toxic, and 54 of the 83 peptides were confirmed to be non-allergic, with the remaining 29 suspected of being allergenic and 31 to be predicted to have good water solubility. Further the molecular docking was used to evaluate the potent dihydropteroate synthase (DHPS) inhibitors. On the basis of ligand binding energy, 17 predicted AMPs were selected and then analyzed by AutoDock tools. Among the 17 AMPs, 3 AMPs were predicted as high-potent antimicrobial. Based on these findings, in silico investigations reveal that proteins of goat milk are a potential source of AMPs. These peptides can be synthesized and improved for use in the food sector. PRACTICAL APPLICATIONS: Goat milk is regarded as a high-quality milk protein source. According to this study, goat milk protein is a possible source of AMPs, and therefore, most important AMPs can be synthesized and developed for use in the food sector.

In silico identification of antidiabetic and hypotensive potential bioactive peptides from the sheep milk proteins-a molecular docking study.

An in silico approach was used for hydrolysis of sheep milk proteins (α-s1, α-s2, ß-casein, κ-Cn, α-lactalbumin, and ß-lactoglobulin) by gastrointestinal enzymes in order to generate bioactive peptides (BAPs) that can inhibit ACE and DPP-IV. Sheep milk proteins showed higher similarity with goat milk proteins. These data were acquired via the Clustal Omega tool to perform sequence alignment analysis. The BIOPEP-UWM database was used to examine the ability of sheep milk protein sequences to generate BAPs, which included a description of their potential bioactivity as well as the frequency of fragments with specified activities. Using the "Enzyme(s) action" tool (BIOPEP-UWM), digestive enzymes pepsin, trypsin, and chymotrypsin, and three enzyme combinations were selected to computationally hydrolyze milk proteins for obtaining information about ACE and DPP-IV inhibitory peptides. Other online programs were used to test potential peptides for bioactivity, toxicity, and physicochemical properties. BAPs produced from PTC-hydrolyzed proteins were analyzed using a peptide ranker, and their inhibitory effects on ACE and DPP-IV were determined using molecular docking. Consequently, the results of molecular docking analysis show that the peptide PSGAW (αS1-Cn f155-159) binds to DPP-IV with binding energy (-8.9 kcal/mol). But in the case of ACE, two potential BAPs were selected: QPPQPL (ß-Cn f161-166) and PSGAW. These two BAPs revealed a higher binding affinity for ACE with a binding energy of -9.8 kcal/mol. Thus, the results showed that sheep milk proteins were a promising source of antidiabetic and hypotensive peptides. However, experimental and pre-clinical studies are necessary to assay their therapeutic effects. PRACTICAL APPLICATIONS: Sheep milk proteins are known as a high-quality milk protein resource. Effective enzymatic hydrolysis of sheep milk proteins can release bioactive peptides and also release potential ACE and DPP-IV inhibitory peptides. This in silico study specifies a theoretical root for sheep milk proteins as a novel source of potential bioactive peptides and may offer guidance for invitro hydrolysis of proteins for the production of bioactive peptides valuable for human consumption.

Novel role of xanthine oxidase-dependent H2O2 production in 12/15-lipoxygenase-mediated de novo lipogenesis, triglyceride biosynthesis and weight gain.

12/15-lipoxygenase (12/15-LOX) plays an essential role in oxidative conversion of polyunsaturated fatty acids into various bioactive lipid molecules. Although 12/15-LOX's role in the pathophysiology of various human diseases has been well studied, its role in weight gain is controversial and poorly clarified. Here, we demonstrated the role of 12/15-LOX in high-fat diet (HFD)-induced weight gain in a mouse model. We found that 12/15-LOX mediates HFD-induced de novo lipogenesis (DNL), triglyceride (TG) biosynthesis and the transport of TGs from the liver to adipose tissue leading to white adipose tissue (WAT) expansion and weight gain via xanthine oxidase (XO)-dependent production of H2O2. 12/15-LOX deficiency leads to cullin2-mediated ubiquitination and degradation of XO, thereby suppressing H2O2 production, DNL and TG biosynthesis resulting in reduced WAT expansion and weight gain. These findings infer that manipulation of 12/15-LOX metabolism may manifest a potential therapeutic target for weight gain and obesity.

A validation study of early warning system in high-risk pregnant women.

High-risk obstetric patients have chances of deterioration which can be detected by any early warning score. This study was aimed to assess the suitability of the Obstetrics National Early Warning System (ONEWS) for the pregnant women. This prospective study was conducted on 500 high-risk pregnant women attending a tertiary care teaching hospital. The ONEWS charts were plotted for each of them. The primary outcome measure was composite adverse maternal outcome (CAMO) in the form of one or more among mortality, severe maternal morbidity and intensive care unit admissions. Of the 500 women who participated, 200 (40%) had a score ≥3 (triggered an intervention). The CAMO among the triggered group [59.5% (n=119)] was significantly higher compared to that in the non-triggered group [13.3% (n=40) (P=0.001)]. The area under the receiver operating characteristic curve was 0.800 (95% confidence interval 0.752-0.847). The sensitivity of the ONEWS in predicting CAMO was 74.8 per cent, specificity 76.2 per cent, positive predictive value 59.5 per cent and negative predictive value 86.7 per cent at a cut-off score of 3. ONEWS appears to be a useful tool for predicting adverse maternal outcomes in high-risk pregnant women.

Revisiting the plant growth-promoting rhizobacteria: lessons from the past and objectives for the future.

Plant beneficial rhizobacteria (PBR) is a group of naturally occurring rhizospheric microbes that enhance nutrient availability and induce biotic and abiotic stress tolerance through a wide array of mechanisms to enhance agricultural sustainability. Application of PBR has the potential to reduce worldwide requirement of agricultural chemicals and improve agro-ecological sustainability. The PBR exert their beneficial effects in three major ways; (1) fix atmospheric nitrogen and synthesize specific compounds to promote plant growth, (2) solubilize essential mineral nutrients in soils for plant uptake, and (3) produce antimicrobial substances and induce systemic resistance in host plants to protect them from biotic and abiotic stresses. Application of PBR as suitable inoculants appears to be a viable alternative technology to synthetic fertilizers and pesticides. Furthermore, PBR enhance nutrient and water use efficiency, influence dynamics of mineral recycling, and tolerance of plants to other environmental stresses by improving health of soils. This report provides comprehensive reviews and discusses beneficial effects of PBR on plant and soil health. Considering their multitude of functions to improve plant and soil health, we propose to call the plant growth-promoting bacteria (PGPR) as PBR.

Consumption of Probiotic Lactobacillus fermentum MTCC: 5898-Fermented Milk Attenuates Dyslipidemia, Oxidative Stress, and Inflammation in Male Rats Fed on Cholesterol-Enriched Diet.

There is a growing and alarming prevalence that increased serum cholesterol is closely related to increased cardiovascular disease risk. Probiotic consumption could be a safe and natural strategy to combat. Therefore, we sought to examine the cholesterol-lowering potential of co-supplementation of probiotic bacteria Lactobacillus fermentum MTCC: 5898-fermented buffalo milk (2.5% fat) in rats fed cholesterol-enriched diet. Male Wistar rats were divided into three groups on the basis of feed, viz. group 1, fed standard diet (SD); group 2, fed cholesterol-enriched diet (CED); and group 3, fed cholesterol-enriched diet along with L. fermentum MTCC: 5898-fermented milk (CED+LF) for 90 days. At the endpoint, significantly higher levels of serum total cholesterol, low-density lipoprotein cholesterol, triacylglycerols, very low density lipoprotein cholesterol, atherogenic index, coronary artery risk index, hepatic lipids, lipid peroxidation, and mRNA expression of inflammatory cytokines (TNF-α and IL-6) in the liver while significantly lower levels of serum high-density lipoprotein cholesterol and anti-oxidative enzyme activities, catalase, superoxide dismutase, and glutathione peroxidase in the liver and kidney were observed in the CED group compared to the SD group. Compared to the CED group, these adverse physiological alterations were found significantly improved in the CED+LF group. Hence, this study proposes that L. fermentum MTCC: 5898 is a potential probiotic bacteria that can be consumed to tackle hypercholesterolemia. Graphical Abstract ᅟ.

A Comparative Study of Energy Contents in Mosquito Vectors of Malaria and Dengue Prevailing in Jodhpur City (Thar Desert) of Rajasthan State, India.

BACKGROUND: Transmission of malaria and dengue in the desert part of India is mainly caused by Anopheles stephensi and Aedes aegypti respectively. The maintenance and transmission of the pathogens that cause malaria and dengue are dependent on the physiology of the mosquito vectors. We aimed to measure the energy contents in the mosquitoes transmitting malaria and dengue in the desert part of the country. METHODS: Immature stages of mosquitoes were collected from six different larval habitats situated in Jodhpur City of Rajasthan state, India. The immature stages of both the mosquitoes were collected once in fortnightly from each location. Quantitative estimations of the lipid, glucose, and glycogen of the laboratory-reared and field collected An. stephensi and Ae. aegypti were made by spectrophotometric method. The energy contents of the larvae, pupae, females, and males were estimated in triplicates on six different occasions. RESULTS: The lipid content of laboratory-reared larvae, pupae and female mosquitoes of An. stephensi and Ae. aegypti was found to be lower than their conspecific field-collected specimens. Whereas, the glycogen content in the laboratory-reared larvae, pupae and female mosquitoes of An. stephensi and Ae. aegypti was higher than that of their conspecific field-collected specimens. The glucose content in all the stages of the laboratory-reared An. stephensi was lower than their conspecific field-collected specimens except in few cases. CONCLUSION: The higher amount of lipid in field-collected mosquitoes may be because of the availability of food in the natural habitat and adaptation of mosquitoes. Mosquitoes living in desert climate are physiologically better equipped to survive in the desert environment.

Evaluation of anti-diabetic attributes of Lactobacillus rhamnosus MTCC: 5957, Lactobacillus rhamnosus MTCC: 5897 and Lactobacillus fermentum MTCC: 5898 in streptozotocin induced diabetic rats.

Interest in probiotics has grown significantly in the last decades due to their reported nutritional and health promoting effects. The aim of this study is to investigate the therapeutic potential of probiotic fermented milk (PFM) prepared using three different probiotic strains i.e. Lactobacillus rhamnosus MTCC: 5957, Lactobacillus rhamnosus MTCC: 5897 and Lactobacillus fermentum MTCC: 5898; independently or in combination, for treating streptozotocin induced type-1 diabetes in male Wistar rats. Diabetic rats were fed with PFM preparations for 6 weeks and then analyzed for the various biochemical parameters associated. The results indicated that feeding of PFM significantly improved glucose metabolism (fasting blood glucose, glycated hemoglobin, serum insulin), serum inflammation status (tumor necrosis factor-α, and serum interleukin-6), oxidative stress (thiobarbituric acid reactive substance, catalase, superoxide dismutase and glutathione peroxidase activities in liver and kidney), serum lipid profile (total cholesterol, low density lipoprotein-cholesterol, very low density lipoprotein-cholesterol, triglycerides) in diabetic rats. In addition, feeding of PFM has significantly reduced mRNA expression of pepck and g6pase genes that code the key enzymes of gluconeogenesis pathway. The results of this study showed that daily consumption of PFM can be effective in combating of type -1 diabetes and its complications.

Evaluation of potassium solubilizing rhizobacteria (KSR): enhancing K-bioavailability and optimizing K-fertilization of maize plants under Indo-Gangetic Plains of India.

Imbalanced potassium (K) fertilization in agricultural fields has led to considerable negative impacts and remains to be the foremost challenge for maize production in India-Gangetic region. Plant growth-promoting rhizobacteria, particularly potassium solubilizing rhizobacteria (KSR), could serve as inoculants and a promising strategy for enhancement of plant absorption of K hence reducing dependency on chemical fertilizers. Maize seeds were microbiolized for 30 min with KSR suspensions. In the present study, the use of chemical fertilizers along with Agrobacterium tumefaciens strain OPVS10 showed pronounced beneficial effect on growth and yield attributes in maize. There was a significant difference among different parameters studied when varying doses of K and KSR strains were applied. Results showed that the combined application of KSR strain OPVS10 with 100% RDK (recommended dose of K) was most effective in modulating growth, physio-biochemical, and yield attributes in maize thus could be regarded as a promising alternative to mineral K-fertilization. Principal component analysis (PCA) revealed that 100-grain weight and grain yield were the most important properties to improve the sustainable growth of maize. Therefore, these KSR strains have different mechanisms for modulating various activities in maize plants. Results suggested that the synergistic application of KSR strain OPVS10 with 100% RDK can be used for optimized breeding, screening, and nutrient assimilation in maize crop. Hence, this eco-friendly approach may be one of the efficient methods for reducing dependency on chemicals, which pose adverse effects on human health directly and indirectly.

Camel milk ameliorates hyperglycaemia and oxidative damage in type-1 diabetic experimental rats.

This study was designed to assess anti-diabetic potential of goat, camel, cow and buffalo milk in streptozotocin (STZ) induced type 1 diabetic albino wistar rats. A total of 48 rats were taken for the study where one group was kept as non-diabetic control group (8 rats) while others (40 rats) were made diabetic by STZ (50 mg/kg of body weight) injection. Among diabetic rats, a control group (8 rats) was kept and referred as diabetic control whereas other four groups (8 rats each) of diabetic rats were fed on 50 ml of goat or camel or cow or buffalo milk for 4 weeks. All the rats (non-diabetic and diabetic) were maintained on standard diet for four weeks. STZ administration resulted in enhancement of glucose, total cholesterol, triglyceride, low density lipoprotein, HbA1c and reduction in high density lipoprotein in plasma and lowering of antioxidative enzymes (catalase, glutathione peroxidase and superoxide dismutase) activities in pancreas, kidney, liver and RBCs, coupled with enhanced levels of TBARS and protein carbonyls in pancreas, kidney, liver and plasma. OGTT carried out at the end of 4 week milk feeding indicated that all milks helped in early maintenance of glucose level. All milks reduced atherogenic index. In camel milk fed diabetic group, insulin concentration enhanced to level noted for non-diabetic control while goat, cow and buffalo milk failed to restore insulin level. HbA1c level was also restored only in camel milk fed diabetic group. The level of antioxidative enzymes (catalase, GPx and SOD) in pancreas enhanced in all milk fed groups. Camel milk and to a reasonable extent goat milk reduced formation of TBARS and PCs in tissues and blood. It can be concluded that camel milk ameliorates hyperglycaemia and oxidative damage in type-1 diabetic experimental rats. Further, only camel milk completely ameliorated oxidative damage in pancreas and normalised insulin level.

Common Mechanism of Cross-Resistance Development in Pathogenic Bacteria Bacillus cereus Against Alamethicin and Pediocin Involves Alteration in Lipid Composition.

To understand the mechanism of development of cross-resistance in food pathogen Bacillus cereus against an antimicrobial peptide pediocin and antibiotic alamethicin, the present study was designed. Pediococcus pentosaceus was taken as a source of pediocin, and it was purified by ammonium sulphate precipitation followed by cation exchange chromatography with 14.01-fold purity and 14.4 % recovery. B. cereus strains alamethicin-resistant strains (IC50 3.23 µg/ml) were selected from sensitive population with IC50 2.37 µg/ml. The development of resistance in B. cereus against alamethicin was associated with decrease in alamethicin-membrane interaction observed by in vitro assay. Resistant strain of B. cereus was found to harbour one additional general lipid as compared to sensitive strain, one amino group lacking phospholipid and one amino group containing phospholipid (ACP). In addition, ACP content was increased in resistant mutant (29.7 %) as compared to sensitive strain (14.56 %). The alamethicin-resistant mutant B. cereus also showed increased IC50 (58.8 AU/ml) for pediocin as compared to sensitive strain (IC50 47.8 AU/ml). Cross-resistance to pediocin and alamethicin in resistant mutant of B. cereus suggested a common mechanism of resistance. Therefore, this understanding could result in the development of peptide which will be effective against the resistant strains that share same mechanism of resistance.

Highly Specific Culture-Independent Detection of YGNGV Motif-Containing Pediocin-Producing Strains.

A novel method based on (1) initial microbiological screening and (2) a highly specific PCR is described for selection of strains expressing YGNGV motif-containing pediocin. Initial screening is carried out using spot on the lawn assay for selection of acid-free, hydrogen peroxide (H2O2)-free and secreted heat-stable inhibitory activity producing strains. This is followed by highly specific PCR for amplification of 406-bp fragment using forward primer: 5'-tggccaatatcattggtggt-3' targeting signal peptide sequence of pediocin structural gene and reverse primer: 5'-ctactaacgcttggctggca-3' encoding N-terminus of immunity gene. The assay was validated with Pediococcus pentosaceus NCDC273 and Pediococcus acidilactici NCDC252 using (1) digestion of amplified 406-bp fragment with HindIII restriction enzyme-producing two restriction fragments of expected sizes (227 and 179 bp), (2) nucleotide sequencing of 406-bp fragment from both strains found these pediocins identical to pediocin PA-1/AcH and (3) identification of both pediocins as pediocin PA-1 at protein level using RP-HPLC. The assay was used for screening six strains (3 pediococci, 2 lactobacilli and an Enterococcus faecium) producing acid-free, hydrogen peroxide (H2O2)-free and secreted heat-stable inhibitory activity. This resulted in the detection of three new strains (P. pentosaceus NCDC35, E. faecium NCDC124 and Lactobacillus plantarum NCDC20) producing YGNGV motif-containing pediocins.