Complete genome sequencing and strain characterization of a novel marine Bacillus velezensis FTL7 with a potential broad inhibitory spectrum against foodborne pathogens.

Bacillus velezensis FTL7 which exhibited potent antimicrobial peptide producing capacity was isolated from a marine sediment sample of the West Coast region, South India, and characterized through experimental and genomic analysis approaches. FTL7 showed potential antimicrobial activity against a broad range of foodborne pathogenic bacteria like Listeria monocytogenes Scott A, Bacillus cereus (ATCC 11778), Salmonella Typhimurium (MTCC 1251), Staphylococcus aureus (ATCC 25923), and Escherichia coli (MTCC 443). It also exhibited strong inhibitory activity against Kocuria rhyzophila (ATCC 934) and Bacillus subtilis subsp. spizizenii (ATCC 6633). Phylogenetic analysis by 16S rRNA gene sequence showed that Bacillus velezensis FTL7 was closely related to B. velezensis LBUM288 (GenBank accession number MG461457) with 100% identity. Whole-genome sequencing of the strain FTL7 was carried out using Illumina sequencing technology to get a better insight into the mechanisms of controlling pathogens by FTL7. The strain FTL7 has a chromosome size of 3849,077 bp with a GC content of 46.56%. The genome consists of 3635 coding sequences, 64 RNA, 59 tRNAs, 5 ncRNAs, and 69 pseudogenes. The presence of genes responsible for the synthesis of non-ribosomal peptides and bacteriocins was identified through genome annotation. Thus, many Bacillus strains, including B. velezensis, have been demonstrated as excellent producers of antimicrobial substances.

Production of egg white hydrolysate by digestion with pineapple bromelain: optimization, evaluation and antioxidant activity study.

Chicken egg white is known to be an excellent source of good quality proteins to make hydrolysate with potential bioactive properties. Enzymatic digestion is a well-known method to produce protein hydrolysates; however, the type of enzyme determines the bioactive potential of the protein hydrolysates due to difference in their catalytic specificity. In this study, process optimization, production and evaluation of whole egg white protein hydrolysate (WEWPH) using pineapple bromelain through the Box-Behnken design were carried out. The design experiment (r 2 = 0.9557) displayed a significant (p < 0.01) effect of pH of egg white (9.0), hydrolysis time (24 h), and enzyme/substrate ratio (3.2 unit/g substrate) on hydrolysis and to form bioactive WEWPH. Antioxidant activity of the WEWPH was confirmed by DPPH radical scavenging assay. Gel filtration chromatography, SDS-PAGE and FTIR spectroscopy analysis of WEWPH revealed the digestion of egg white and the integrity of WEWPH in terms of secondary structure. The WEWPH exhibited strong scavenging activities of DPPH (EC50 = 238.3 µg/ml), ABTS ABTS (EC50 = 54.9 µg/ml), peroxyl (EC50 = 391.6 µg/ml) and superoxide radicals. The WEWPH also displayed reducing power and singlet oxygen quenching activity. These results reveal that the bioactive WEWPH could be a promising ingredient in health food and nutraceuticals. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-021-05188-0.

Malabar sole (Cynoglossus macrostomus) skin as promising source of type I acid and pepsin solubilized collagens with potential bioactivity.

Fish skin is one of the major non-edible by-products formed during fish processing. This investigation focused on the sustainable valorization of Malabar sole (MS) skin for collagen, which can be utilized as potential alternative of mammalian collagen. Acid and pepsin solubilized collagen (ASC and PSC) were successfully isolated from MS skin with a yield (%, dry weight basis) of 49.5 ± 0.6 and 67.6 ± 0.5, respectively. The isolated collagens were characterized by SDS-PAGE, UV-absorption, DSC, SEM, FTIR spectroscopy, etc., analysis. Both collagens were characterized as type I by SDS-PAGE and the well preserved triple helical structure by FTIR and UV absorption analysis. Denaturation temperature (°C) of the MS skin collagens confirmed by DSC analysis was 33.67 (ASC) and 33.38 (PSC). Both collagens showed high solubility in acidic pH and low NaCl level, and also exhibited a comparatively high degree of fibril-forming capacity. Antioxidant potential of the isolated collagens was confirmed by DPPH (31.4-34.6% at 1.5 mg) and peroxyl (64.6-68.3% at 0.3 mg) radical scavenging assays and observed a dose dependent manner activity. Overall, the results suggested the possibility of using the MS skin as a potential substitute source of realistic type I collagen and also help to reduce issues of fish processing discards. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at (10.1007/s13197-021-04996-8).

Optimization of conditions for isolation of high quality chitin from shrimp processing raw byproducts using response surface methodology and its characterization.

Chitin is one of the most abundant bioactive biopolymer on earth. It is commercially extracted from seafood processing crustacean shell byproducts by harsh thermochemical treatments. The extraction conditions, the source and pretreatment of raw material significantly affect its quality and bioactivity. In this investigation response surface methodology (RSM) has been applied to optimize and evaluate the interaction of variables for extraction of high quality chitin from shrimp processing raw byproducts. Variables such as, concentration of HCl (%, v/v) 4.5 (for wet) and 4.9 (for dry), reaction time 3 h, solid liquid ratio of HCl (w/v) 1:5.5 (for wet) and 1:7.9 (for dry) with two treatments achieved >98 % demineralization of shrimp byproduct. Variables such as, concentration of NaOH 3.6 % (w/v), reaction time 2.5 h, temperature 69.0 ± 1 °C, solid liquid ratio of NaOH 7.4 (w/v) and two treatments accomplished >98 % deproteinization of demineralized byproducts. Significant (p ≤ 0.05-0.001) interactive effects were observed between different variables. Chitin obtained in these conditions had residual content (%, w/w) of ash <0.4 and protein <0.8 and the degree of N-acetylation was >93 % with purity of >98 %. In conclusion, the optimized conditions by RSM can be applied for large scale preparation of high quality chitin from raw shrimp byproduct.

Application of chitosan for improvement of quality and shelf life of table eggs under tropical room conditions.

The current investigation was conducted to study the effectiveness of chitosan coating in preserving the internal quality of table eggs stored under tropical room conditions of 32 ± 1 °C and 60-70 % r. h. Internal, physical and microbiological quality of eggs coated with chitosan was evaluated during 5-week storage at different temperature (22 ± 1 and 32 ± 1 °C). Chitin was extracted from shrimp processing raw byproducts and deacetylated to high quality chitosan. The prepared chitosan was analyzed for its characteristic properties. The chitosan with a viscosity of 2206 mPa.S was used to prepare the coating solution. The weight loss, Haugh unit, and yolk index values suggested that coating of eggs with shrimp α-chitosan increased the shelf life of eggs by almost 4-week at 22 ± 1 °C and 3-week at 32 ± 1 °C compared with controls (non chitosan coated and acetic acid coated) eggs. Three-time repeated coating was more effective in preserving the internal quality and preventing weight loss than with single-time coating of chitosan on egg. Therefore, three-time coating of eggs with 2206 mPa.S chitosan offer a protective barrier for preserving the internal quality of eggs stored at tropical room conditions and concomitantly prevent contamination with microorganisms.

Extracellular chitin deacetylase production in solid state fermentation by native soil isolates of Penicillium monoverticillium and Fusarium oxysporum.

Extracellular chitin deacetylase production by native soil isolates of Penicillium monoverticillium CFR 2 and Fusarium oxysporum CFR 8 in solid state fermentation (SSF) using commercial wheat bran (CWB) and shrimp processing by-products (SPP) as solid substrate has been studied. P. monoverticillium produced maximum chitin deacetylase activity of 547.7 ± 45 and 390.2 ± 31 units/g initial dry substrate (U/g IDS) at 96 h of incubation in CWB and SPP media, respectively. While, F. oxysporum produced maximum chitin deacetylase activity of 306.4 ± 22 U/g IDS at 72 h of incubation in CWB medium and 220.1 ± 20 U/g IDS at 120 h of incubation in SPP medium. Along with chitin deacetylase, P. monoverticillium and F. oxysporum produced other chitin degrading enzymes such as endo-chitinase and ß-N-acetylhexosaminidase. P. monoverticillium produced maximum activity (U/g IDS) of endo-chitinase 4.6 ± 0.20 at 120 h incubation and ß-N-acetylhexosaminidase 82.6 ± 03 at 120 h incubation in CWB medium. While, F. oxysporum produced maximum activity (U/g IDS) of endo-chitinase 7.8 ± 0.20 at 144 h incubation and ß-N-acetylhexosaminidase 38.3 ± 02 at 120 h incubation in CWB medium. Production of extracellular chitin deacetylase by P. monoverticillium CFR 2 and F. oxysporum CFR 8 in SSF is being reported for the first time.

Extraction optimization, partial purification, and characterization of sialoglycoproteins from Labeo rohita roes.

In India, fish roes are generally considered worthless garbage and disposed of without recovering the valuable molecules, creating environmental and disposal problems. The present investigation aimed to optimize the extraction conditions, partial purification, and characterization of sialoglycoproteins (RRSGP) from Labeo rohita (rohu) roes. RSM generated optimum conditions for maximum RRSGP (70.49 %) extraction, which were 1.25 M NaCl, 1:32.5(w/v) solid-to-liquid ratio, 47.5 °C temperature, and 3 h time. Further, sialoglycoproteins from RRSGPs were partially purified, and result revealed that obtained peak-1 (PRRSGP) using QFF anion exchange chromatography exhibited higher glycoprotein and sialic acid content (p < 0.05). SDS-PAGE pattern of PRRSGP presented dominant bands of 97 kDa and 27 kDa glycoproteins. FTIR spectrum of PRRSGP confirmed the presence of glycated proteins. HPLC analysis revealed that PRRSGP consists of Neu5Ac. Furthermore, ß-elimination reaction elucidated that PRRSGP contained N-glycosidic linkage. PRRSGP exhibited tyrosine and glutamate as primary amino acids. Glycan part of PRRSGP presented mannose and N-acetyl galactosamine as dominant neutral and amino sugar, respectively. Furthermore, PRRSGP exhibited antioxidant activity with EC50 value for DPPH (8.79 mg/ml) and ABTS (2.21 mg/ml). Besides, RRSGP displayed better protein solubility, foaming, and emulsion properties. Therefore, rohu roes are potential source of sialoglycoproteins that can be recovered and used as bio-functional ingredients in food and nutraceutical applications.

Anti-listerial peptides from a marine Bacillus velezensis FTL7: production optimization, characterizations and molecular docking studies.

Antimicrobial peptides (AMPs) with potent anti-listerial activity were characterized from a novel marine Bacillus velezensis FTL7. A Box-Behnken statistical experimental design was used to study the combined impact of culture conditions on the production of AMPs by B. velezensis FTL7. The conditions optimized by statistical experimental design were 34.5 °C incubation temperature, 23 h incubation time, and 7.6 initial pH of the medium. AMP purification was performed by ammonium sulphate fractionation and butanol extraction followed by reversed-phase C18 solid-phase extraction. Tricine-SDS-PAGE analysis revealed a peptide with a molecular mass of ~ 6.5 kDa in an active AMPs fraction, whereas the mass spectrometry (MS) analysis showed the presence of AMPs in the mass range of 1-1.6 kDa, along with a 6.5 kDa peptide. Both MS and MS/MS analysis confirmed the AMPs as lipopeptides including surfactin, fengycins and iturin A and a circular bacteriocin amylocyclicin. The minimum inhibitory concentration of these AMPs against L. monocytogenes Scott A was 2.5 µg/mL. Further, the in-silico docking studies showed that the AMPs from B. velezensis FTL7 have high binding energy and stable binding patterns towards L. monocytogenes target proteins. Thus, this new combination of AMPs can serve as an effective food bio-preservative. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-024-03944-5.

Effect of defatted Lagenaria siceraria (Molina) Standley seed flour as a fat replacer on physicochemical, technological, and sensory properties of beef patty.

Defatted Lagenaria siceraria seed flour (DLSSF) was obtained from defatted seed cake, dried, and ground through a sieve of 500 µm and characterized. A 2 × 4 factorial design (two flour hydration rates and four fat substitution rates) was used to produce a low-fat beef patty by replacing fat with DLSSF. Beef kidney fat was used to formulate the control sample. Chemical, physical, technological, sensory, and nutritional characteristics of low-fat beef patties manufactured were evaluated. DLSSF contains mainly protein. As fat replacers, DLSSF induces a significant increase in the pH of the raw and cooked patty, the moisture and protein contents, the cooking yield, the cohesion, chewiness, springiness, and lightness of the cooked beef patty with fat substitution rate. There is a decrease in fat content, total calories, water retention capacity, hardness, and redness of the cooked patty with a fat substitution rate. From the sensory analysis, the substitution of fat improves the acceptability of samples. Based on the overall parameters analyzed, DLSSF containing 60% water can be used to produce low-fat beef patty by replacing fat at 100%. From these results, hydrated DLSSF could be an effective method to solve the problems of noncommunicable diseases related to animal fat consumption.

Facial phenotype at different ages and cardiovascular malformations in children with Williams-Beuren syndrome: a study from India.

Williams-Beuren syndrome (WBS) is one of the microdeletion syndromes associated with distinct facial features, characteristic behavior phenotype (overfriendly behavior), congenital heart disease, and other malformations. Clinical features in WBS are age dependent. It is important to be aware of variable age dependent phenotype, especially facial phenotype due to its crucial role in diagnosis. Here we describe the facial phenotype of WBS at different ages (3 months to 15.1 years) and congenital heart malformations in 27 patients FISH positive for 7q11.23 microdeletion.

Enhanced degradation of α-chitin materials prepared from shrimp processing byproduct and production of N-acetyl-D-glucosamine by thermoactive chitinases from soil mesophilic fungi.

Soil isolates of mesophilic Penicillium monoverticillium CFR 2, Aspergillus flavus CFR 10 and Fusarium oxysporum CFR 8 were cultivated in solid state fermentation (SSF) using wheat bran solid medium supplemented with α-chitin in order to produce chitinolytic enzyme. Under SSF cultivation, maximum enzymes (U/g IDS) production was 41.0 (endo-chitinase) and 195.4 (ß-N-acetylhexosaminidase) by P. monoverticillium, 26.8 (endo-chitinase) and 222.1 (ß-N-acetylhexosaminidase) by A. flavus and 13.3 (endo-chitinase) and 168.3 (ß-N-acetylhexosaminidase) by F. oxysporum after 166 h of incubation. The crude endo-chitinase and ß-N-acetylhexosaminidase derived from A. flavus and F. oxysporum revealed optimum temperature at 62 ± 1°C, but the enzymes from P. monoverticillium showed optimum temperature at 52 ± 1°C for maximum activity. Several fold increase in endo-chitinase and ß-N-acetylhexosaminidase activities in the crude enzymes preparation was achieved after concentrating with polyethylene glycol. The concentrated crude chitinases from P. monoverticillium, A. flavus and F. oxysporum, respectively yielded 95.6, 96.6 and 96.1 mmol/l of N-acetyl-D: -glucosamine (GlcNAc) in 48 h of reaction from colloidal chitin. While, the crude enzyme preparations of P. monoverticillium, A. flavus and F. oxysporum produced 10.11, 6.85 and 10.7 mmol/l of GlcNAc respectively, in 48 h of reaction from crystalline α-chitin. HPLC analysis of colloidal chitin hydrolysates prepared with crude chitinases derived from P. monoverticillium, A. flavus and F. oxysporum revealed that the major reaction product was monomeric GlcNAc (~80%) and a small amount of (GlcNAc)(4) (~20%), indicating the potential of these enzymes for efficient production of GlcNAc from α-chitin.

Biodegradation of shrimp processing bio-waste and concomitant production of chitinase enzyme and N-acetyl-D-glucosamine by marine bacteria: production and process optimization.

A total of 250 chitinolytic bacteria from 68 different marine samples were screened employing enrichment method that utilized native chitin as the sole carbon source. After thorough screening, five bacteria were selected as potential cultures and identified as; Stenotrophomonas sp. (CFR221 M), Vibrio sp. (CFR173 M), Phyllobacteriaceae sp. (CFR16 M), Bacillus badius (CFR198 M) and Bacillus sp. (CFR188 M). All five strains produced extracellular chitinase and GlcNAc in SSF using shrimp bio-waste. Scanning electron microscopy confirmed the ability of these marine bacteria to adsorb onto solid shrimp bio-waste and to degrade chitin microfibers. HPLC analysis of the SSF extract also confirmed presence of 36-65 % GlcNAc as a product of the degradation. The concomitant production of chitinase and GlcNAc by all five strains under SSF using shrimp bio-waste as the solid substrate was optimized by 'one factor at a time' approach. Among the strains, Vibrio sp. CFR173 M produced significantly higher yields of chitinase (4.8 U/g initial dry substrate) and GlcNAc (4.7 µmol/g initial dry substrate) as compared to other cultures tested. A statistically designed experiment was applied to evaluate the interaction of variables in the biodegradation of shrimp bio-waste and concomitant production of chitinase and GlcNAc by Vibrio sp. CFR173 M. Statistical optimization resulted in a twofold increase of chitinase, and a 9.1 fold increase of GlcNAc production. These results indicated the potential of chitinolytic marine bacteria for the reclamation of shrimp bio-waste, as well as the potential for economic production of chitinase and GlcNAc employing SSF using shrimp bio-waste as an ideal substrate.

Solid state fermentation production of chitin deacetylase by Colletotrichum lindemuthianum ATCC 56676 using different substrates.

Production of extracellular chitin deacetylase by Colletotrichum lindemuthianum ATCC 56676 under solid substrate fermentation was studied. The suitability of shrimp shell chitin waste (SSCW) and commercial wheat bran (CWB) was evaluated for maximal enzyme production. CWB medium (pH 6.4 ± 0.2) supplemented with chitosan favoured maximal chitin deacetylase yield of 460.4 ± 14.7 unit/g initial dry substrate (U/g IDS) at 96 h as compared to maximal yield of 392.0 ± 6.4 U/g IDS at 192 h in SSCW medium (pH 8.7 ± 0.2) at 25 °C incubation temperature and 60% (w/w) initial moisture content of medium. Along with chitin deacetylase, C. lindemuthianum ATCC 56676 produced maximum endo-chitinase (0.28 ± 0.03 U/g IDS at 144 h) and ß-N-acetylhexosaminidase (0.79 ± 0.009 U/g IDS at 192 h) in CWB medium and 0.49 ± 0.05 U/g IDS of endo-chitinase at 264 h and 0.38 ± 0.04 U/g IDS of ß-N-acetylhexosaminidase at 96 h of incubation in SSCW medium. SEM studies indicated the difference in the morphology of mycelia and hyphae of C. lindemuthianum ATCC 56676 when grown on different solid substrates. Production of chitin deacetylase by SSF is being reported for the first time.

Indian Guidelines for Indications and Timing of Intervention for Common Congenital Heart Diseases: Revised and Updated Consensus Statement of the Working Group on Management of Congenital Heart Diseases. Abridged Secondary Publication.

JUSTIFICATION: A number of guidelines are available for management of congenital heart diseases from infancy to adult life. However, these guidelines are for patients living in high income countries. Separate guidelines, applicable to Indian children, are required when recommending an intervention for congenital heart diseases, as often these patients present late in the course of the disease and may have co-existing morbidities and malnutrition. PROCESS: Guidelines emerged following expert deliberations at the National Consensus Meeting on Management of Congenital Heart Diseases in India, held on 10th and 11th of August 2018 at the All India Institute of Medical Sciences, New Delhi. The meeting was supported by Children's HeartLink, a non-governmental organization based in Minnesota, USA. OBJECTIVES: To frame evidence based guidelines for (i) indications and optimal timing of intervention in common congenital heart diseases; (ii) follow-up protocols for patients who have undergone cardiac surgery/catheter interventions for congenital heart diseases. RECOMMENDATIONS: Evidence based recommendations are provided for indications and timing of intervention in common congenital heart diseases, including left-to-right shunts (atrial septal defect, ventricular septal defect, atrioventricular septal defect, patent ductus arteriosus and others), obstructive lesions (pulmonary stenosis, aortic stenosis and coarctation of aorta) and cyanotic congenital heart diseases (tetralogy of Fallot, transposition of great arteries, univentricular hearts, total anomalous pulmonary venous connection, Ebstein anomaly and others). In addition, protocols for follow-up of post surgical patients are also described, disease wise.

Characterization and application of a native lactic acid bacterium isolated from tannery fleshings for fermentative bioconversion of tannery fleshings.

Lactic acid bacteria (LAB) species isolated from limed and delimed tannery fleshings (TF) were evaluated for their fermentation efficiency and antibacterial property. The native LAB isolates efficiently fermented TF and resulted in a fermented mass with antioxidant properties, indicating their potential for effective eco-friendly bioconversion of TF. From among the LAB isolated, a proteolytic isolate showing better antimicrobial spectrum and reasonably good fermentation efficiency was identified as Enterococcus faecium HAB01 based on various biochemical and molecular tests. This isolate afforded a better degree of hydrolysis (81.36%) of TF than Pediococcus acidilactici (54.64%) that was previously reported by us. The bacteriocin produced by E. faecium was found to be antagonistic to several human pathogens including Listeria, Aeromonas, Staphylococcus and Salmonella. Further, E. faecium HAB01 bacteriocin was thermostable and had a molecular weight of around 5 kDa, apart from being stable at both acidic and alkaline conditions. The bacteriocin was unstable against proteases.

Severe mitral regurgitation: a misleading presentation.

Anomalous left coronary artery arising from the pulmonary artery (ALCAPA) is a rare congenital anomaly commonly presenting in infancy with severe congestive cardiac failure due to myocardial ischemia. We present a two and half year old child who presented with severe mitral regurgitation and near normal left ventricular function and turned out to be having ALCAPA. High pulmonary artery pressures prevented coronary steal into the pulmonary arteries and preserved LV function (near noprmal) at presentation.

Cardiac mucormycosis with T-cell immunodeficiency.

We present a case of a 2 year old apparently healthy child who presented with fever and mass on the mitral valve. Excision histopathology of the mass revealed mucormycosis. After 4 months, she had CNS embolisation with recurrence of cardiac lesion when investigations revealed associated T-cell immunodeficiency.

Indian guidelines for indications and timing of intervention for common congenital heart diseases: Revised and updated consensus statement of the Working group on management of congenital heart diseases.

A number of guidelines are available for the management of congenital heart diseases (CHD) from infancy to adult life. However, these guidelines are for patients living in high-income countries. Separate guidelines, applicable to Indian children, are required when recommending an intervention for CHD, as often these patients present late in the course of the disease and may have coexisting morbidities and malnutrition. Guidelines emerged following expert deliberations at the National Consensus Meeting on Management of Congenital Heart Diseases in India, held on August 10 and 11, 2018, at the All India Institute of Medical Sciences. The meeting was supported by Children's HeartLink, a nongovernmental organization based in Minnesota, USA. The aim of the study was to frame evidence-based guidelines for (i) indications and optimal timing of intervention in common CHD; (ii) follow-up protocols for patients who have undergone cardiac surgery/catheter interventions for CHD; and (iii) indications for use of pacemakers in children. Evidence-based recommendations are provided for indications and timing of intervention in common CHD, including left-to-right shunts (atrial septal defect, ventricular septal defect, atrioventricular septal defect, patent ductus arteriosus, and others), obstructive lesions (pulmonary stenosis, aortic stenosis, and coarctation of aorta), and cyanotic CHD (tetralogy of Fallot, transposition of great arteries, univentricular hearts, total anomalous pulmonary venous connection, Ebstein's anomaly, and others). In addition, protocols for follow-up of postsurgical patients are also described, disease wise. Guidelines are also given on indications for implantation of permanent pacemakers in children.

Guidelines for the management of common congenital heart diseases in India: A consensus statement on indications and timing of intervention.

INTRODUCTION: A number of guidelines are available for management of congenital heart diseases from infancy to adult life. However, these guidelines are for patients living in high-income countries. Separate guidelines, applicable to Indian children, are required when recommending an intervention for congenital heart diseases, as often these patients present late in the course of the disease and may have co-existing morbidities and malnutrition. PROCESS: Guidelines emerged following expert deliberations at the National Consensus Meeting on Management of Congenital Heart Diseases in India, held on the 10th and 11th of August, 2018 at the All India Institute of Medical Sciences. OBJECTIVES: The aim of the study was to frame evidence-based guidelines for (i) indications and optimal timing of intervention in common congenital heart diseases and (ii) follow-up protocols for patients who have undergone cardiac surgery/catheter interventions for congenital heart diseases. RECOMMENDATIONS: Evidence-based recommendations are provided for indications and timing of intervention in common congenital heart diseases, including left-to-right shunts, obstructive lesions, and cyanotic congenital heart diseases. In addition, protocols for follow-up of postsurgical patients are also described.

Recovery of carotenoids from ensilaged shrimp waste.

A method for fermentation of shrimp waste was standardized using a statistically designed experiment, with respect to three variables namely, levels of glucose and starter culture and time of fermentation. The optimized levels for achieving the desired pH was 20.5% glucose, 19.5x10(4)cells/g of starter culture and fermentation time of 70h. Recovery of carotenoids from fermented and acid ensiled shrimp waste was assessed during 75 days of storage. Acid ensilaging resulted in the reduction of solvent extraction yield of carotenoids from 43.09 to 26.76 microg/g by the end of 75 days of storage. The yield of oil extracted carotenoids was higher in both types of silage at the end of 75 days storage compared to the initial yield, being 31.30 microg/g in fermented silage and 26.18 microg/g in acid silage. The results indicated the usefulness of fermentation as a method for stabilization and recovery of carotenoids in the shrimp waste.