Title: Environment and Sustainability: Transforming Biomass into a Resource Fibre waste management systems: opportunities, costs and technology advisory resources”

Responsible: Dr. Pedro de Oliva Neto

Department of Biological Science – São Paulo State University- UNESP - Assis – SP - Brazil Av. Dom Antonio 2100 – Assis –SP CEP 19806-390 - poliva@assis.unesp.br

Project: Development of research with waste of sisal (Agave sisalana Perrine) – Pharmacological properties.

Research carried out by UNESP in collaboration with:

- Common Fund of Food and Agriculture Organization of the United Nations - Sindycate of industries of vegetables fibers of Bahia State SINDIFIBRAS – Bahia – Brazil- Secretary of Science, Technology and Invention of State of Bahia - Brazil

Agreement signed by SINDIFIBRAS and FUNDUNESP (Fundation of São Paulo State University) in May 2010.

• Researchers :

• Prof. Dr. Pedro de Oliva Neto - Coordinator (Biotechnology, Microbiology)

• Prof. Dra. Lucinéia dos Santos (Toxicology, Pharmacology)• Dra Ana Flavia Azevedo Carvalho (Microbiology , Biochemistry)• Dr. Regildo Márcio Gonçalves da Silva – (Pharmacology - Phytochemistry)• Dra. Edislane Barreiros de Souza – (Genetic – Mutagenicity)• Dra. Isabel Cristina Cherici (Cell Biology – Reproduction)• Dr. Alvimar José da Costa (Insect Control – mits in animals) • Dr. Carlos Amadeu Leite de Oliveira (Insect Control in agriculture)• Dr. Antonio Carlos Maringoni (Phytopatology – Bacteriology) • Dr. Daniel J. de Andrade (Insect Control in agriculture) • Dra. Dejanira de Franceschi de Angelis (Ecotoxicity)• Dr. Edson Luiz Lopes Baldin (Insect control in agriculture and animals) • New member: Dr. Marcelo Mattos Dinato ( Doctor specialized in pre-

clinical and clinical tests).

Introduction

The Agave - plant originated from Yucatan - Mexico. Precious fiber - for the manufacture of various utensils such as carpets, ropes, twine, marine cables, bags, etc, and also for natural food source (feed) and alcoholic beverages like a international tequila.  .

Agave sisalana Perrine (sisal) - the main hard fibre produced worldwide. It was introduced in Brazil in 1903 (the largest exporter of this fibre in the world) In Brazil - cultivation in states of Bahia (93.5%), Paraiba and Rio Grande do Norte, in a poor and semi arid region - small producers (family labor). The sisal fibre - 100 million dollars is exported from Brazil and generate more than half million jobs.

General Goal of this project To study the technical use of sisal waste in order to develop new commercial products contributing to sisal workers.

Results of the Project 1. Data Process of Pulping and pressing Sisal leaves.___________________________________________________________________ Sisal waste %(dried w/w) Wet Mass %___________________________________________________________________ Total cut leaves (1 m in length): 1645 Total weight leaves: 580.0 kg 100% Total wet fibres1: 65.2 kg 11.2% Total full wet mucilage 14.6% 503.0 kg 86.7% Loss 2.1% Total liquid waste (sisal juice)2 5.0% 176.8 kg 30.5% Total solid waste (EMT)3 18.1% 326.2 kg 56.2% Liquid from process full mulcilage (EMS) 2.5% 378.0 kg ___________________________________________________________________ 1. the pulping process of the leaves was 1 h. 2. after pressing and filtering the

mucilage 3. Solid waste from full mucilage after pressing and filtering the full mucilage

The pulping machine The sisal leaves

• The pressing machine is very simple and economically viable (very low cost) .

• Press machine

• Chemical composition of sisal juice

•

Chemical determinations Results

Total caloric value 31.31 kcal/100g

Total Carbohydrates 7-12 g/100g

Fiber 0 g/100g

Protein 0.31 g/100g

Fat 0.23 g/100g

Mineral Material 1.03 g/100g

Moisture 91-95 g/100g

Sodium (Na) 97.71 mg/kg

Flowchart of the treatments of mucilage from sisal leaves in

some products studied.

Preliminary cost for some products of sisal waste.__________________________________________________ main products operations workers production cost in 8 h. (reais) ___________________________________________________ _______________ EMS - Liquid from FM pressing and filtration 2 3038 l. R$ 0.01/l OJ - Original juice pressing 2 1414 l. R$ 0.02/l DP - Dried Precipitate centrifugation, drying 1 4.45kg R$ 9.73/kg AP - Acid Precipitate heating, acidification and centrifugation 1 4.95kg R$ 14.92/kg___________________________________________________________________ DP – R$ 15.00/1 worker (8 h)+ R$ 28.28 (cost of 1,414 l juice) AP - R$ 15.00/ 1 worker (8 h)+ R$ 28.28 (cost of 1,414 l juice) + cost of H2SO4 34 l x R$ 0.90/l = R$ 30.6 total = R$ 73.88

Obs. Several costs is not considered (energy, depreciation, investiments, etc), but it is calculating by an expert.

2. Biological tests

• The preliminar study was divide in three categories depending on: • the results of analysis , degree of technical difficulties, costs and time demanded to become a

commercial product.

• Category A – Products with high chances of commercial application.

• 1) Agent against mites on citrus (orange, lemon, tangerine).2) Cream for mycoses.3) Shampoo antifungal (antidandruff) 

• Malassezia ssp.

Class B - Products with intermediate chances of commercial application.1) Antioxidant for use in cosmetic creams .2) Action on inhibition of ticks.

3) Anti-inflammatory Action 4) Anti-parasitic nematodes of plants 5) Action contraceptive

Class C - Products that technically has no recommendation for further studies - (due to showed very weak or non existent action for the target) .

1. Insecticidal action of sisal juice 2. Antibacterial action against against whitefly (attack leaves of legumes) Xanthomonas (phytopatogenic bacteria)

Toxicological studies of Sisal waste were:• Ecotoxicity (Daphnia) • Male Reproductive toxicity in rats • Mutagenicity• Median Lethal Dose (dosage to kill 50% of the rats)

Tests of antifungic activities of sisal waste .

Based on MIC (Minimal Inhibitory Concentration) test of several products formulated by sisal waste . MIC Test – Macrodilution of antimicrobial compounds

Microorganisms used:

Candida albicans (ATCC 10231) Malassezia pachydermatis (ATCC 14522)

Cream for C. albicans 60gShampoo antimycotic 100 ml

Cream for skin mycosis 60g

Lotion for nail mycosis30ml

Mass of Sisal dried precipitate for bottle

8.4 g 1.7g 1.03g 0.51g

Cost of dried precipitate for bottle1 R$ 0.082 R$ 0.016 R$ 0.01 R$ 0.005

Cost of formulation more bottle 2 R$1.13 R$ 2.18 R$ 1.0 R$ 1.00

Total cost (only the formula and bottle)

R$ 1.21 R$ 2.20 R$ 1.01 R$ 1.00

Trade value R$ 16.38 (Nystatin) R$ 39.00 (cetoconazol) R$ 18.00 (cetoconazol) R$ 142.00 (12 ml)

Cost for the formulation of creams, lotion and shampoo from sisal waste (Dried Precipitate).

Obs. Not included others cost like energy, governament taxes, transport and employers for the cosmetic industry

Class A – Acaricide Effect of Sisal waste on mite Brevipalpus phoenicis, (vector of citrus leprosis)

Experiments with direct effect of Agave waste on Brevipalpus phoenicis in 12 h.______________________________________________________________________ Product Concentr. Death efficiency vol/ha2 cost/ha (R$)______________________________________________________________________________Original juice (5% solid) 100% 94-100% 4000 l. 80.00 2.7 time conc.juice (12% solid) 100% 100%Dried Acid precipitate 2% 100% 80 kg 1194.00 Dried precipitate 0.2% 25-46%EMT - Solid from Mucilage (18% sol.) 2% 37-46% EMS – Liquid from Mucilage (2.5 % sol.) 80% 100% 3200 l. 64.00 cyhexatin (Sipcatin) 0,05%1 100% 2 l. 300.00 Control (distilled water) _____________________________________________________________________

Sipcatin is used 50 mL to 100 L of water or 1 liter of commercial product for 2,000 liters (or 0.05%) of water and 2 liters per hectare – R$150/l. 2. volum spend per Hectare.

Flowchart of Pilot plant for waste sisal processing

Category - Class B

1.Effect against nematodes . Family Strongylidae gen.Strongylus

- Represents the main source of losses for Sheep farmers around the world. Other animals are infected (ex. horses).

Method - Egg counts of nematodes (gastrointestinal strongyles) per gram of feces (EPG) in sheep after treatment with the Acid Precipitate of sisal juice (dosage: 0.02 g/ml - 1 ml/ 10 kg of animal, de animal, administered orally.

Results : Reduction in average - 68.1% of number of eggs per gram of feces of the animal.

Conclusion: These results are preliminary but promising, since only one treatment was used and only one dose. A more detailed tests are recommended. PS. We do not test all products of sisal waste (EMT, EMS, Dried Precipitate)

Reproductive parameters of female ticks Rhiphicephalus

The most important arthropod in veterinary medicine as result of economic losses and health problems.

Methodology: the reproductive parameters of female ticks were evaluated after treatment with the juice of sisal. The parameters analyzed were:% reduction in oviposition,% reduction of hatchability, reproductive efficiency.

Products tested: Pure and diluted juice, Acid Precipitate (2%)

RESULTS:

% reduction in oviposition = results were not significant at different treatm. % reduction of hatchability = Best result was with sisal juice (75% diluted in water) – 47% reduction of hatchability and efficience of 57% in the ticks control.

Conclusion: The efficience in the control of ticks was limited however it is recommended that a new research is conducted, in more depth. 

Evaluation of antioxidant potential of concentrated juice of Agave sisalana (Agavaceae).

Methodology based on the stable free radical DPPH and absortion of H + quantified by Absorbance at 517 nm (spectrophotometry). 2.7 times concentrated juice was used in several dilution._____________________________________

Concentrated juice Antioxidant activity Conclusion: ____________________________________ Sisal juice has a potential for use

0.05 % 3.31% as a natural antioxidant for cosmetic

0.1 % 4.75% or other products depending on

0.5 % 13.9% more applied studies.

1.0 % 24.5%

2.5 % 57.8%

5.0 % 68.1%

10% 75.0%

_______________________________________

Sisal waste toxicity

• Acid precipitated from sisal juice showed:

• Spermicide action on rats if ingested.• Ecotoxicity is considered high (superior of oil refinery,

so not recommended to discharge into rivers. • Not mutagenic in dosages up to 100 mg/kg, superior

dosages was not tested.• We need to test Liquid from Mucilage and Dried

precipitate toxicity.

Final Conclusion

This research especially with the insecticide and antimycotic activity, respectively, with the products Liquid from full mulcilage and Dried Precipitate from sisal juice needs to be continued.

The new steps will require a pilot plant for processing greater amount of full mucilage to new tests in animals (field tests) and clinical tests in humans.

We believe this research points to a promising technology development from

sisal waste in order to obtain economic value and generating wealth and welfare for the workers of sisal.

Thank You