Incidences of Bronchial Asthma have been raised in recent decades due to increased industrialization and pollution. This miserable condition can be compared with Tamaka Shwasa in Ayurveda. Modern synthetic drugs will provide instant relief in these cases, but are tend to develop a number of adverse drug reactions. Knowing this, the current suffering population is looking towards few remedies from other systems of medicines, that are comparatively safe and provide better relief. Shirisha [Albizzia lebbeck Benth] is a drug with multi-dimensional activities emphasized in Ayurveda for different disease conditions. Considering this, two types of Shirishavaleha (confection of Shirisha) were prepared by Kwatha (decoction) of Twak (bark) and Sara (heartwood) of Shirisha to evaluate its comparative efficacy in Tamaka Shwasa (bronchial asthma). The results were assessed in terms of clinical recovery, symptomatic relief and pulmonary function improvement. A significant increase in Hb and considerable decrease in total eosinophil count, AEC and ESR were observed. The study revealed that Shirishavaleha can be used as an effective drug in bronchial asthma.
Avaleha; Sara; Shirisha; Tamaka Shwasa; Twak
The purpose of the present study was to evaluate the anti-inflammatory activity of Shirishavaleha prepared from two different parts of Shirisha (Albizia lebbeck Benth.), viz. the bark (Twak) and the heartwood (Sara). The activity was screened in the carrageenan-induced rat paw edema model in albino rats. The raw materials were collected and authenticated in the university and the trial formulations were prepared by following standard classical guidelines. Randomly selected animals were divided into four groups of six animals each. The test drugs were administered orally at a dose of 1.8 g/kg for 5 days. Phenylbutazone was used as the standard anti-inflammatory drug for comparison. Between the two different test samples studied, the formulation made from heartwood showed a weak anti-inflammatory activity in this model while that made from the bark produced a considerable suppression of edema after 6 h. It appears that the bark sample would be preferable for clinical use.
Albizia lebbeck; anti-inflammatory activity; Avaleha
The immunomodulatory activity of Shirishavaleha prepared from two different parts of Shirisha (Albizia lebbeck Benth), i.e., Twak (Bark) and Sara (Heartwood) as main ingredients was evaluated for humoral antibody formation and cell-mediated immunity in established experimental models. The study used Wistar rats of either sex weighing 200 ± 40 g, while the test drug was administered orally at a dose of 1.8 g/kg. Hemagglutination titer and body weight were recorded to assess effects on humoral immunity; immunological paw edema was assessed for cell-mediated immunity. Shirishavaleha prepared from heartwood shows significant enhancement in antibody formation, attenuation of body weight changes, and suppression of immunological paw edema, while Shirishavaleha prepared from bark shows weak immunomodulatory activity. The study therefore concludes that Shirishavaleha prepared from heartwood has significant immunomodulatory activity.
Albizia lebbeck; avaleha; hemagglutination titer; immunomodulatory activity
Iron is one among the major metals present in the earth's crust and is essential for sound sustenance of human body. Its deficiency leads to various health ailments. Contemporary medicine advises iron supplements in iron deficiency anemia. Ayurvedic classics also quote significant information about administration of iron. Lauha Kalpas are the unique compound herbo-mineral formulations where iron (Lauha) is used as a major ingredient. Relevant literature (Bhaishajya Ratnavali, Charaka Samhita, Rasendra Sara Samgraha etc.) reviewed to gather information about Lauha Kalpas. Critical analysis of these Lauha Kalpas reveals that ancient seers administered iron in a better acceptable form. Unlike popular understanding these are not only Khalviya preparations; but Churna (powders), Avaleha (confectionaries), Rasakriya (solidified decoctions), and Putapaka (incinerated) form of preparations are also found. Apart from solid dosage forms, semisolid dosage forms mentioned in classics are very much useful. Unfortunately most of the formulations are not found in the market. Hence Pharmaceutical firms may bring these unique dosage forms in to the market to supply the healthcare needs of the community. It is interesting that iron preparations are used in Ayurveda in different medical conditions apart from anemia (Pandu). This leaves a scope for further researches on different dosage forms of iron and their indications.
Ayurveda; Bhasma; Hematinics; Iron; Lauha Kalpa; pharmaceutics; Rasaushadhi
Vasa (Adhatoda vasica Linn.) is a well known and easily available drug in almost all the seasons. Easy availability of any drug gains popularity among physicians as well as pharmaceuticals and this is the reason why almost every Kalpana of Vasa is found described in the Ayurvedika text. The different dosage forms of Vasa like Kvatha, Avaleha, Sneha, and Sandhana have been used for the treatment of Shwasa Roga. A number of research studies have been performed on different formulations of Vasa and its effect on Shwasa Roga. Therefore, a review study has been carried out on the Vasa extract, Vasa Avaleha (prepared from Svarasa and Kvatha), Vasa Ghrita, Vasarishta, and Vasakasava on Shwasa Roga, to know which formulation is better. It was found in the review that Vasa Ghana, Vasa Ghrita (1), and Vasa Avaleha have shown good results on Tamaka Shwasa.
Ghana (extract); Avaleha; Shwasa; Asava; Arishta; Tamaka Shwasa; Adhatoda vasica
Avaleha, which are generally elegant preparations, can be used for both preventive and curative purposes. Many research works have been carried out on different Avalehas. A number of studies with different Avalehas and their respective clinical efficacy in Tamaka Shwasa (Bronchial Asthma) have been carried-out at Department of Rasashastra and Bhaishajya Kalpana, IPGT & RA, Jamnagar. A review has been carried out of such works to know the better one. In the work, it has been found that highly significant (P<0.001) results on Shwasakastata were found in all the groups, except in the Kamsa Haritaki Avaleha (B) group which was insignificant (P<0.02).
Avaleha; Brimhana; Shwasa
Vasa (Adhatoda vasica) is used to treat the diseases such as Shwasa, Kasa and Raktapitta in different dosage forms like Swarasa, Avaleha and Ghrita. Although the Avaleha Kalpana is not available in Brihattraya, but Gada Nigraha by Aacharya Sodhal and Bhava Prakasha have described its use in the form of Avaleha to treat the diseases of Respiratory System.
The objective of this study is to compare the efficacy of two types of Vasa Avaleha prepared with either ‘Swarasa’ or ‘Kwatha’ of Vasa during their preparation. The outcomes were assessed on the basis of relief in subjective symptoms and certain hematologicalparameters.
Total 35 patients were enrolled for the study. Both the groups showed highly significant results on cardinal symptoms like frequency, intensity and duration of Shwasa (dyspnoea), Kasa (coughing), Peenasa (rhinitis) with maximum percentage in Vasa Avaleha (Swarasa). The formulations also shows a insignificant decrease in haematocrit values which includes Neutrophil, Eosinophil, Lymphocyte count and TL.STL. Overall Vasa Avaleha (Swarasa) shows maximum percentage of improvement than the other group.
Mayur Puccha bhasma (Calx of peacock feather), an Ayurvedic animal product prepared from peacock feather with different methods mentioned in Siddhayogasangraha, Bhaishajya Samhita. It is mainly indicated in Chardhi, Hikka, and Swasa. Till today no research work has been done on Mayur Puccha bhasma prepared by two different methods and exact pharmacopeial standards are also not available. The present study was aimed to assess the Physico chemical components of Mayur Puccha bhasma prepared by Ghee flame and Gajaputa method & to lay down the Pharmacopeial standards for the assessment of its standard qualities.
Mayur Puccha bhasma was prepared by two classical procedures, one by burning on ghee flame (Sample A) second by giving four Gajaputas (Sample B) and finished products were subjected to various Physico chemical studies to find out Ash value, PH value, Sp gravity, Moisture content, Preliminary organic analysis and Gravimetric analysis.
Both the bhasmas exhibited marked difference in colour, moisture content and % of inorganic compounds. The bhasma prepared by Gajaputa method contains essential and beneficial inorganic elements, electrolytes in more quantity with less moisture content, neutral pH.
Sample B is having less moisture (1%) content which may contribute for increasing its stability and having high ash value as it consists of inorganic elements in more quantity and indicates that bhasma prepared properly. The neutral pH of Sample A and B indicates no significance difference in their pH value. Sample B contains Copper, Iron, Zinc, Sodium, Potassium, Calcium, Magnesium, Manganese, Aluminium, in more quantity than Sample A but in a permissible amount. As Mayur Puccha bhasma prepared by Gajaputa method contains essential and beneficial elements, having natural pH and free from steroids may contribute as one of the usefully and safe alternative medicine in Chardhi, Hikka, Swasa.
The use of Neutraceuticals has drastically risen in recent years. Dr Stephan De Felice coined the term Neutraceuticals from “nutrition” and “pharmaceutical” in 1989. Related terms are “functional food” and “dietary supplement”. In Ayurvedic pharmaceutics there are some secondary preparations like Avaleha Kalpana (Medicated semisolid preparation), Asavarista Kalpana (fermented preparation), Sneha Kalpana (Medicated fatty preparation), Ksheerapaka Kalpana (Medicated milk preparation) etc. which can be correlated with Neutraceuticals. In this paper “Neutraceuticals” and “Avaleha Kalpana” have been correlated and discussed.
Neutraceuticals; Functional food; Avaleha Kalpana
Ayurvedic concept is of the opinion that Tamaka Shwasa (Bronchial Asthma) is a Yapya Vyadhi. The etiopathogenesis, signs, and symptoms of Tamaka Shwasa may be correlated with Bronchial Asthma. Each child reacts differently to the factors that trigger asthma and treated symptomatically. Asthma is the most common chronic allergic disorder in childhood and third leading cause of hospitalization under the age of 15 years. As it is a Kapha-Vata predominant disorder, Ayurvedic medicine may help to decrease the recurrence, improve immunity, and check symptoms naturally. With this aim, a clinical study was undertaken on two groups for duration of 6 weeks. The drugs Bharangyadi Avaleha and Vasa Avaleha were given orally, separately in both the groups. All the patients were kept under strict dietary control during the treatment. The observation on effect of therapy was encouraging and showed less recurrence.
Bharangyadi Avaleha; childhood asthma; Tamaka Shwasa; Vasa Avaleha
The present study is carried out to evaluate the immuno-modulatory activity of Vacha Dhatryadi Avaleha in albino rats. Vacha Dhatryadi Avaleha was prepared by classical method and evaluated for humoral antibody formation and cell-medicated immunity in established experimental models. Test formulation was administered at the dose of 900 mg/kg and parameters like hemagglutination titer, ponderal changes, histopathology of immunological organs and immunological paw edema were recorded. Vacha Dhatryadi Avaleha significantly enhanced antibody formation and moderately suppressed the immunological edema. The present study concludes that Vachadhatryadi Avaleha has immunopotentiating activity.
Cell mediated immunity; hemagglutination titer; Vachadhatryadi
Dhatryarishta has been described for the first time in Charaka Samhita, in the context of Panduroga. The same reference is available in Chakradatta, Bhaishajya Ratnavali, and Sahastrayogam too. Generally Dhatri Swarasa [Amalaki (Emblica officinalis Gaertn.)] is used in the preparation of Dhatryarishta as per classical reference, but fresh Amalaki is not available in every season, so in the present study, Amalaki Kwatha (decoction) is used instead of Swarasa. A total of 13 batches of Dhatryarishta were prepared, out of which nine batches were prepared with Dhatri Swarasa and four batches with Dhatri Kwatha. For Dhatryarishta prepared by using Dhatri Swarasa, three methods were applied and in Dhatryarishta prepared by using Dhatri Kwatha two methods were applied. The study revealed that Dhatryarishta could only be prepared by using Amalaki Swarasa as quoted in the classics and not by using Amalaki Kwatha.
Dhatri; Dhatryarishta; Kwatha; Panduroga; Swarasa
Tamaka Shvasa which has been mentioned in Ayurvedic classics shares multiple similarities with Bronchial Asthma. Symptom of breathlessness is the main complaint in Bronchial Asthma which can be assessed objectively by Pulmonary Function Test (PFT).The assessment of respiratory function is now a routine part of clinical practice. The expiratory flow rates- Forced Expiratory Volume in first second (FEV), Forced Vital Capacity (FVC) and Peak Expiratory Flow Rate (PEFR) are assessed by an Electronic Spirometer (Kent, England). Six weeks treatment with a compound preparation of herbs including - Sati (Hedichum spicatum, Rose), Puskaramoola (Innula racemosa, Linn), and Amalaki (Emblica officinalis, Gaertn) powder showed a significant effect of increase in Pulmonary Function values. The mean grade score plus standard deviation before trial of FEV, FVC, and PEFR were 62.6±15.06, 2.03±o.53 and 189±44.05 respectively. After six weeks of treatment with Puskaramooladi choorna FEV, FVC and PEFR showed highly significant results with values 63.45±15.9, 2.81±0.33 and 199.6±41.58 respectively. Puskaramooladi choorna can be used as one of the potent medicine in the treatment of the Bronchial Asthma.
Ayurveda refers to bronchial asthma as Tamaka Swasa and it is well explained in Charaka Samhita. It contributes several modalities of the treatment for Swasa roga(asthma). Among all modalities of treatment, polyherbal combinations are said to be well-accepted, safe and effective in asthma. A study was carried out in 40 patients of either sex in between the age of 15-65 years to assure the clinical response of Padmapatradi yoga in bronchial asthma (Tamaka Swasa) at P.G. department of Kayachikitsa, D.G.M. Ayurvedic Medical College, Gadag, Karnataka. The sum total properties of Padmapatradi yoga is tikta katu rasa, laghu and tikna guna (light and penetrating properties), ushna virya (hot potency) and vatakaphagna (decrease vata and kapha dosa) Padmapatradi yoga is effective in increased peak expiratory flow rate, breath holding time, and reduces the absolute eosinophil count of studied cases and also found statistically highly significant at p<0.001 level. The drug is quite safe and acts as a bronchodilator, antihistaminic and anti-inflammatory.
Breath holding time; bronchial asthma; dysponea; forced expiratory volume; Padmapatradi yoga; peak expiratory flow rate; Tamaka Swasa; vital capacity
Tamaka Shvasa is a type of Shvasa Roga associated with difficulty in breathing as a result of which the patient prefers to sit in bed to get relief from his discomfort. Movement of air through Pranavaha Srotas is hampered in this disease resulting in the cry of organ heading toward complete failure for want of air. Tamaka Shvasa is well known for its episodic and chronic course which comes under the life-threatening disease. It is analogous to bronchial asthma due to similarity in symptoms, pathogenesis, onset, causes, and precipitating factors. In this study, 40 patients of Tamaka Shvasa were registered and randomly divided into two groups, out of which 31 patients completed the treatment. In Group A, Shvasahara Leha (5 g twice a day) was given for 2 months, while in Group B Vasa Haritaki Avaleha (5 g twice a day) was given for 2 months and follow-up was done for one month in both groups. The effects of therapy in both groups were assessed by a specially prepared proforma. Diagnosis was done by adult asthma diagnosis questionnaire and differential diagnosis with COPD (Chronic obstructive pulmonary disease) was done by differential diagnosis questionnaire as both these conditions are overlapping. The results of the study indicate that the Vasa Haritaki Avaleha provided better relief than Shvasahara Leha in Tamaka Shvasa.
Bronchial asthma; Shvasahara Leha; Tamaka Shvasa; Vasa Haritaki Avaleha
To verify the antidiabetic potential of stem bark of Albizzia lebbeck (A. lebbeck) and seeds of Mucuna pruriens (M. pruriens) using various in vitro techniques.
The plant extracts were studied for their effects on glucose adsorption, diffusion amylolysis kinetics and glucose transport across yeast cells.
Both the plant extracts adsorbed glucose and the adsorption of glucose increased remarkably with an increase in glucose concentration. No significant (P≤0.05) differences were observed between the adsorption capacities of A. lebbeck and M. pruriens. In amylolysis kinetic experimental model the rate of glucose diffusion was found to increase with time from 30 to 180 min, and both the plant extracts demonstrated significant inhibitory effects on movement of glucose into external solution across dialysis membrane as compared to control. The retardation of glucose diffusion by A. lebbeck extract was significantly higher (P≤0.05) than M. pruriens. These effects were reflected with higher glucose dialysis retardation index values for A. lebbeck than M. pruriens. The plant extracts also promoted glucose uptake by yeast cells. The rate of uptake of glucose into yeast cells was linear in all the 5 glucose concentrations used in the study. M. pruriens extract exhibited significantly higher (P≤0.05) activity than the extract of A. lebbeck at all concentrations.
The results verified the antidiabetic potential of A. lebbeck and M. pruriens. The hypoglycemic effect exhibited by the extracts is mediated by increasing glucose adsorption, decreasing glucose diffusion rate and at the cellular level by promoting glucose transport across the cell membrane as revealed by simple in vitro model of yeast cells.
Hypoglycemic; In vitro; Glucose diffusion
To investigate antimicrobial and bronchodialator effect of hydroalcholic extract of polyherbal drug Shirishadi containing Shirisha (Albezzia lebbeck), Nagarmotha (Cyprus rotandus) & Kantakari (Solanum xanthocarpum).
Antimicrobial activity was evaluated by disc diffusion method and MIC, MBC, MFC were calculated by micro dilution method. Hydroalcholic extract of this preparation was investigated for its phytochemical analysis, phenol and flavonoid were determined by spectrophotometric method and in vivo bronchodilator effect was analysed by convulsion time.
The phytochemical tests revealed presence of alkaloids, anthraquinones, carbohydrates, flavonoids, saponins and tannins. The antimicrobial result showed the MIC of 6.25 mg/mL against Staphylococcus aureus and 12.5 mg/mL for Escherichia coli and 12.5 mg/mL against remaining bacteria tested, with strong antifungal activity. The maximum inhibition zone is found against Pseudomonas aeruginosa with MIC 16 mg/mL. Drug showed significant bronchodilator effect with 27.86% & 36.13% increase in preconvulsion time of guinea pigs pretreated with 100 & 200 mg/kg body weight of extract.
The study reveals that the extracts possess antibacterial activity and antifungal activity in a dose dependent manner. This antimicrobial property may be due to presence of several saponins, further studies are highly needed for the drug development.
Shrishadi-polyherbal drug; Phytochemical; Phenolic & flavonoid content; Candida species; Bronchodilator
Shukti is an important component of Sudha Varga, which is considered as the latest class in the field of Rasa Shastra. Two types of Shukti have been mentioned in Rasa Shastra texts i.e. Jala Shukti and Mukta Shukti according to the availability. In present study, an attempt has been made to develop a standard manufacturing procedure (SMP) of Jala Shukti Bhasma and Mukta Shukti Bhasma. Five batches of Jala Shukti Bhasma and Mukta Shukti Bhasma were prepared and standardization was attempted by maintaining batch manufacturing records of individual batches. During pharmaceutical procedures like Shodhana, Bhavana, Marana, etc. due care of temperature, its duration, percentage of weight gain or loss and the cost factor of the end product, etc. were considered. The average weight loss observed was 12.08 g i.e. 2.42% and 14.62 g i.e. 2.92% during Jala Shukti and Mukta Shukiti Shodhana respectively. Average weight loss found was 38.94 g i.e. 7.79% in Jala Shukti Bhasma while in Mukta Shukti Bhasma, it was 35.24 g i.e. 7.05%. At the end of the pharmaceutical procedure, it was found that Mukta Shukti Bhasma is 2.8 times costlier than Jala Shukti Bhasma.
Bhasma; Jala Shukti; Marana; Mukta Shukti; Shodhana
Mamajjaka (Enicostemma littorale Auct. non Bl) is a well known folklore medicine frequently used for the treatment of Madhumeha (diabetes mellitus). There is no direct reference available for its antihyperglycaemic activity in Ayurvedic classics. Considering this, a study is planned towards developing pharmaceutical standardization of Mamajjaka Ghana. In this study, five batches of Mamajjaka Ghana were prepared and findings were systematically recorded to maintain the Standard Operating Procedure (SOP). An average of 14.78% Ghana was obtained. The physico-chemical parameters, qualitative test for various functional groups, quantitative estimation of total alkaloids, HPTLC profile, heavy metal analysis and microbial overload were carried out of Mamajjaka Ghana.
Enicostemma littorale; Ghana; Kwatha; Mamajjaka
In ancient days, Physicians having the comprehensive knowledge of Bhaishajya Kalpana, used to prepare the drugs themselves to treat their patients. So there was no doubt in obtaining genuine drug with desired therapeutic effect. But in recent years, the growing population and their life style, industrialization etc have forced physicians to depend on market preparations. As such we find the necessity of standardization of these preparations. The quality assessments of a drug, which is a chemical irrespective of the system is possible by ‘Thin Layer Chromatographic technique’ using known Chemical constituents as reference standards. A herbal preparation ‘Kutajarishta’, has been standardized by using this technique and the significance of the findings is discussed.
Three types of tamra bhasmas separately prepared by using the media of mercury, Solanum xanthocarpum and sulphur were tested in patients of tamaka svasa. Results suggest that the process of making tamra bhasma with sulphur is superior to others due to several reasons.
Modern medicine is rapidly developing medications that provide instant relief from symptoms, but the hazards of beta agonists, toxicity of xanthenes and complications of corticosteroids prompt the search for alternative modalities in the management of bronchial asthma. The present work evaluates easily available, inexpensive and non-controversial herbal drugs as well as Pranayama in the management of bronchial asthma.
For this clinical study 60 patients have been divided into 3 groups. 1st Group was treated with trial drug and 2nd Group with standard drug. The 3rd Group was treated with trial drug and Pranayama. All the groups have equal number of patients i.e. 20 each. The short-listed patients were diagnosed on the basis of a detailed questionnaire including the history, clinical examination as well as laboratory investigations. The diagnosis was further confirmed using spirometer. Diagnosis has been made according to bronchial reversibility test i.e. 15% increase in FEV1 after 15 minutes of two puffs of a adrenergic agonist. Subjective as well as objective parameters were adopted for assessing the response of trial drugs graded in to 0, 1, 2, and 3. Patients were advised to practice the anulom vilom pranayama BD, 10m.
Results were discussed on basis of % improvement, standard deviation, p value, p value for eg. N BT, AT, % imp, S.D., t, p, result, - Group III, 18, 2.05, 1.33, 35.12, 0.46, 6.65< 0.001 HS
Group I & III shows significant improvement in clinical symptoms whereas Group III patients also showed marked improvement in PULMONARY FUNCTION TESTS (FEV1, FVC, PEFR). It was observed that anulom vilom pranayama has a definite additive effect with standard drug therapy in the treatment of Tamak swasa (bronchial asthma) as shown by the subjective and objective assessment.
Beauty is not in the face; Beauty is a light in the heart. In our Ayurveda it is stated that Physical, Mental and Spiritual beauty as a combined unit projects the cosmetic sense of Ayurveda (Ch.Su 1/41). According to Ayurveda healthy skin is the result of overall condition of individuals and prescribes numerous skin care treatments that need to be pursued at every stage of life. Ayurveda determines beauty by Prakruti(body constitution), Sara (structural predominance), Samhanana (compactness of body), Twak (skin complexion), Pramana (measurement) and Dirghayu lakshana (symptom of long life). Ayurveda cosmetology started with garbhini paricharya, Dinacharya, Ratricharya and Ritucharya with practice of medicinal herbs and minerals. In day-to-day life, knowingly or unknowingly we exposing lots of chemicals /toxins to our body, those spoiling our natural beauty silently, this same concept is explained in Agada tantra under concept of Dhooshivisha (Su.Ka 2/33) and Garaviasha.
1. Literature review of concept related to cosmetology in relation to Agada Tantra.2. Analysis and Interpretation of mode of action of Agada yoga's in relieving the symptoms of Twak vikaras.
1. Specific Agada yogas like Bilwadi Agada , Dhooshivishari Agada, Moorvadi choorna, Paarantyadi Keram shows best preventive and curative action in twak vikaras.2. Some of the Agadas like Moorvadi choona (A.hrudaya) lepas have enhansess the internal and external beauty.
Agada tantra is not only a branch of Ayurveda it deals with management of acute and chronic diseases but also a added benefit of being used for cosmetology.
This paper deals with the standardisation of Eladhi choornam . It is a compound preparation of 6 ingredients along with sugar, viz 1. Nagakesara, 2. Maricham, 3. Twak, 4. Ela, 5. Sunti, 6. Pippali. Chromatographic technique was used for the detection of the ingredients. Different solvent systems were developed for each ingredients and spots were visualized in Iodine vapours. Presence of spots with identical Rf values in the single drugs and finished product indicate the presence of the single drugs in the finished product the parameters evolved in this stud can be considered viable for this preparation.
Mutation of sarA in Staphylococcus aureus results in a reduced capacity to form a biofilm, but the mechanistic basis for this remains unknown. Previous transcriptional profiling experiments identified a number of genes that are differentially expressed both in a biofilm and in a sarA mutant. This included genes involved in acid tolerance and the production of nucleolytic and proteolytic exoenzymes. Based on this we generated mutations in alsSD, nuc and sspA in the S. aureus clinical isolate UAMS-1 and its isogenic sarA mutant and assessed the impact on biofilm formation. Because expression of alsSD was increased in a biofilm but decreased in a sarA mutant, we also generated a plasmid construct that allowed expression of alsSD in a sarA mutant. Mutation of alsSD limited biofilm formation, but not to the degree observed with the corresponding sarA mutant, and restoration of alsSD expression did not restore the ability to form a biofilm. In contrast, concomitant mutation of sarA and nuc significantly enhanced biofilm formation by comparison to the sarA mutant. Although mutation of sspA had no significant impact on the ability of a sarA mutant to form a biofilm, a combination of protease inhibitors (E-64, 1-10-phenanthroline, and dichloroisocoumarin) that was shown to inhibit the production of multiple extracellular proteases without inhibiting growth was also shown to enhance the ability of a sarA mutant to form a biofilm. This effect was evident only when all three inhibitors were used concurrently. This suggests that the reduced capacity of a sarA mutant to form a biofilm involves extracellular proteases of all three classes (serine, cysteine and metalloproteases). Inclusion of protease inhibitors also enhanced biofilm formation in a sarA/nuc mutant, with the combined effect of mutating nuc and adding protease inhibitors resulting in a level of biofilm formation with the sarA mutant that approached that of the UAMS-1 parent strain. These results demonstrate that the inability of a sarA mutant to repress production of extracellular nuclease and multiple proteases have independent but cumulative effects that make a significant contribution to the biofilm-deficient phenotype of an S. aureus sarA mutant.