Development and Application of Microbial Insecticides for Biological Control of Mosquito in Gaza Strip

A project proposal submitted by the Islamic University-Gaza

to

Science & Technology Office

Islamic Development Bank Group

Contact

Dr. abdelraouf A. Elmanama

Medical technology department

P.O Box 108, Gaza, Gaza Strip

PNA

1. breif analysis

1.1 Present status

Inhabitants of Gaza Strip are annually suffering from mosquito outbreaks during the summer and early fall seasons. This predominance of mosquitoes is mainly due to the availability of suitable site for their breeding. Stagnant water pools are ideal breeding sites. These pools can be formed from rain water runoff or from irrigation runoff.

The major breeding sites of mosquito in Gaza Strip are the inefficiently operated wastewater treatment plants, Wadi Gaza (Fig. 1) and open disposal of sewage. Other breeding sites include; standing water in water storage tanks, abandoned wells, opened sewerages, leakage from damaged water distribution systems, stagnant water located in animals watering baths, wet agricultural soil, collected water in trash like jars, cans, bottles and tires, accumulated water after plant irrigation, water accumulated at the roofs of buildings, leaky faucets and irrigation ponds. Due to the poor hygiene and sanitary conditions, these breeding sites of mosquito are abundant in Gaza Strip, which are further contribute to mosquito's outbreaks. A 42 species of mosquitoes including Culex pipiens and C. univittatus were identified in Palestine.

Fig. 1 Photographs illustrating standing water in Wadi Gaza.

Mosquitoes are primarily an annoyance, causing human pain, discomfort, and stress. The major concern however, is the fact that they are vectors of important human and animal diseases.

The currently practiced methods for mosquito control in Gaza include; direct application of oil and insecticides onto water bodies or open burning of adult mosquitoes. Although chemical insecticides have generally been efficacious, their production costs are high and they present environmental concerns. For example, because of their mode of action, they can cause many ecological problems by destroying harmful and harmless and even useful insects. Insecticides contaminate the soil and may end up in groundwater and thus lead to health problems. Additionally, many insecticides are no longer effective due to mosquitoes acquired resistance.

Obviously, there is an urgent need to encourage the development and implementation of cost-effective, environmentally sound alternatives to the currently employed practices for mosquito control.

This year Gaza Strip suffered from shortage of fuel and almost all basic stuff due to the siege imposed by the Israeli forces. One of the consequences of the siege is the lack of the chemical used to eradicate mosquitoes. Instead, most concerned localities were able to purchase Bacillus thuringiensis BT) and used it for the first time as biological insecticides for mosquito control. Since 1981, BTI has become the leading biopesticide in routine control programmes against mosquitoes. It has attracted industry to use it worldwide as an effective weapon against agricultural pests and insect vectors of human diseases. BT are sold in a variety of formulation (liquid, granule or briquette) under a wide variety of trade names such as:

Mosquito Dunks®, VectoBac™, Aquabac™, Bti Briquets™. Worldwide sales of BT dwarf those of any other biopesticide product. Annual sales in the early 1990s were estimated at $100 million, accounting for l-2% of the global insecticide market.

A local production of BT seems appropriate solution for this issue, especially from cheap local raw materials. This project proposal focuses on production of BT from potatoes and chicken feather (both are available locally).

1.2 BT as bio-pesticide

The use of BTI as biological insecticides has resulted in a valuable option to chemical insecticides. BT is a Gram-positive, spore-forming bacterium, producing at least four major protein toxins that are confined to a crystalliferous inclusion body. These toxins have been shown to be highly effective against a wide variety of mosquito species in different climatic. It has been also shown that BT strains can be very specific in their lethal activity against different insect pests while being harmless to mammals, birds or beneficial insects. In addition, food products treated with this insecticide are safe for human or animal consumption. The BT insecticide is also biodegradable and will not accumulate in the environment or cause pollution problems. Accordingly, increasing attention is directed to BT as a viable alternative to chemical pesticides.

1.3 Requirements for Successful Production of BT

Like other microorganisms BT needs a culture media with the following ingredients for growth, reproduction and spore formation: water, a carbon source for biosynthesis and energy, a nitrogen source, mineral elements and other optimal growing factors. Most strains of BT grow best at 30°C under vigorous aeration and a pH level between 6.8 and 7.2.

Fig. 2: The development stages of Bacillus thuringiensis

1.4 Commercial production of BT and economical impact

One of the keys to successful commercialization of BT insecticides is the development of an adequate culture medium. When cultured in appropriate media, vegetative cells sporulate and lyse, releasing spores and parasporal crystals into the medium (Fig. 2).

Current industrial production of BT is conducted by batch liquid fermentation process or submerged fermentation in which the cultures grow dispersed by air in synthetic liquid media at controlled pH and temperature. After harvest, the product is concentrated and either dried or stabilized as a liquid suspension using different preservatives such as sorbitol, sodium benzoate, xylol, etc. , to avoid further growth and germination of the spores.

1.5 Limitation of the industrial production of BT

Although different synthetic culture media for industrial production of BT are proposed for use, their high cost limits their use for the commercial production especially in developing countries. There is therefore a need for an alternative, inexpensive media which will overcome the above-mentioned limitation.

1.6 Proposed ingredients for cheap growth media

Various efforts have been made to evolve efficacious yet media. For example the following ingredients have been suggested in order to reduce the cost of the media for the commercial production of BT bio-insecticides: cheese whey, corn steep liquor, sorter liquor, cotton seed meal, wheat bran, extracts of potatoes, carrots and sweet potatoes, cassava starch, maize, cowpea liquor, fodder yeast, fish meal, cotton seed meal, horse beans, wheat bran, citrus peels seeds of dates, waste products of the coconut oil industrial processes, the by-products of monosodium glutamate production, the cow blood dehusked greengram powder, defatted soybean powder, soluble starch and cane sugar molasses, and bird feather extract.

These ingredients are generally added to synthetic media comprising water, glucose, yeast extract and a plethora of growth enhancing additives such as nitrogen sources, protein sources usually in the form of leguminous seeds, such as peanuts, chick peas, lima beans, horse beans, kidney beans and soya beans, mineral salts such as CaCO3, NaCl, K2HPO4., MgSO4, CaCl2, FeSO4 and CuSO4 and small amounts of some amino acids. Initial results of an unpublished research performed at the Islamic University, showed promising results fron cheap locllay available materials (Chicken feathers and potato extracts). Accordingly, this project proposal focuses on the economic production of BT by using inexpensive available materials, composed mainly of agro-industrial by-products.

2. The Project

2.1 Project summary

This project focuses mainly on the control of mosquitoes in Gaza strip using the biopesticide Bacillus thuringenesis as an alternative for chemical control. The activities of this work will include; the isolation of a local strain of BT in parallel with commercially available BT; optimizing cultural and environmental parameters. Testing and comparing the efficacy of both local and commercial strain will also be evaluated in the laboratory and in the field. The ultimate objective of this project is to commercially produce BT using locally available raw materials (Chicken feathers and Potato). This will allow safer and readily available mean for mosquitoes control.

2.2 Objectives and scope

The aim of this project is to reduce the potential threat of disease transmitted by mosquito. The main objectives of this study include the following:

1- To produce BT by a simple and effective process in order to obtain an active locally-produced biological control agent.

There are two main advantages in promoting development of local production facilities:

(1) Stability: locally produced microbial insecticides avoid lengthy shipping periods and long storage at variable temperatures before the product reaches the consumer.

(2) Formulations: local production provides material for formulations suitable for local environmental conditions

2- To reduce costs of BT production by using inexpensive available materials which make BT useful not only for small scale application but for large-scale commercialization.

3- To optimize the culture conditions to give high cell yield, high crystal protein concentration and high toxicity.

4- To encourage the application of microbial control in environmentally sensitive habitats (Wadi Gaza) to improve habitats for fish and wildlife.

5- To get mosquito control workers and the community acquainted with the basic aspects of biological control through preparation of an educational guide on the biological control of mosquitoes.

6- To enhance the environmental awareness of official institutions and the public with regard to biological control.

7- To develop the scientific and technical capability of local research institutions in conducting studies in the field of biological control.

8- To increase the public awareness concerning the areas where mosquitoes are likely to breed which can significantly reduce the mosquito's population.

To reduce mosquito-induced discomfort, annoyance, and distraction experienced by personnel

2.2 Scope

Local research is essential in order to implement and develop microbial insecticides for mosquito control. This requires laboratory and field scale studies for testing the efficacy of microbial larvicides under the prevalent environmental conditions. Protocols must be developed, standardized, documented and disseminated for the optimum use of microbial insecticides. The availability of qualified personnel and research laboratories will contribute to the success of the project.

Implementation and development of microbial insecticides for control of mosquitoes will directly benefit humans and the environment. Use of microbial insecticides will significantly lessen the use of chemical insecticides and therein reduce potential health hazards to humans and wildlife, and pollution of soil and groundwater.

This project proposal suggests that mosquito biological control strategy should be given a high priority and support. The proposed activities will contribute significantly to the application and development, for the first time in Gaza Strip, of biological control for the noxious mosquitoes.

The proposed study is expected to:

  1. Support the local organizations in the field of biological control through providing them with results on the efficacy of using bacteria on controlling mosquito populations.
  2. Produce educational guide on biological control of mosquitoes by the use of bacteria and other means.
  3. Disseminate knowledge and information gained in the field of mosquito biological control through the media.
  4. Develop the scientific and technical capability of Environmental & Rural Research Center in conducting studies in the field of biological control.

2.3 Project Location

2.3.1 Geographical location

Gaza strip is a narrow piece of land lying in the coast of the Mediterranean Sea. Its position on the cross roads from Africa to Asia made it a target for occupiers and conquerors over the centuries. The last of these was Israel who occupied the Gaza strip from Egyptians in 1967.

The Gaza Strip is situated in the south part of historical Palestine and southeast of the Mediterranean. The Palestinian National Authority (PNA) has gained a political control Gaza Strip and the West bank as a result of the Oslo agreement between Israel and the Palestinian Liberation organization. This report will only deal with the Gaza Strip coastal area (the only coastal area controlled by the PNA). The map is showing the major governorate of the Gaza Strip.

During the British Mandate Period, the area that is presently called Gaza Governorates was formerly part of the Gaza Sub-District of Palestine. It was one of its 18 sub-districts and it includes three towns: Gaza, Khan-Yunis and Majdal, and 54 villages. The area was 1111.5 km2 of which 49.3 km2 were Jewish owned. Gaza with its present area of about 365 km2, and its present borders and delimiting lines, first came into existence 50 years ago in 1948, after the first Israeli-Arab war.

Geographically, Gaza is part of the Palestinian coastal plain in Southwest Palestine, where it forms a long narrow rectangle. Its length is approximately 42 km2; and its width 5.7 kilometers in the Northern section, attaining a maximum, of 12 kilometers at the Southern end.

2.3.1 Demography

It is one of the most densely populated places on earth. Approximately 1.5 million Palestinians live in an area of 365Km. Political developments and economical situation in the region strongly influenced the growth and distribution of population in the Governorates. Gaza Strip have a very young population in comparison to other countries, 50.2 % of the population is 15 years. As much as 21% of the Gaza citizens are 4 years or younger. In addition, Gaza Governorates have an average of nearly 9 persons per household. Moreover, they have a higher average number of children per adult household members in comparison with West Bank and East Jerusalem.

2.3.2 Topography

Elongated ridges and depressions, dry streambeds and shifting sand dunes, characterize Gaza topography. The ridges and depressions generally extend in a NNE- SSW direction, parallel to the coastline. They are narrow and consist primarily of 'Kurkar' sandstone. In the south, these features tend to be covered by sand dunes. Land surface elevations range from mean sea level (msl) to about 10 meters above mean sea level (msl). The ridges and depressions show considerable vertical relief, in some places up to 60m. Surface elevations of individual ridges range between 20m and 90m above msl. The major topographic depressions, filled with alluvial sediments, collect considerable quantities of storm-water. Flooding is common in Rafah and Gaza City during the wet season.

The parallel, Kurkar ridges have been dissected by Wadi Gaza, the largest surface water feature in Gaza. It rarely flows due to numerous water diversion and storage projects upstream in Israel. However, it has historically cut an incised valley with river terraces to the sea. There are two other wadis in Gaza: Wadi Silka near Khan Younis (a fossil river and now a dry wash) and Wadi Halib near Beit Hanoun (a tributary of the much larger Nahal Shiqma (Israeli terminology) that drains to the sea just north of Gaza.

Gaza is essentially a foreshore plain gradually sloping westwards. The quaternary rocks are visible as Kurkar ridges go South West-North East. The ridges have an increasing height towards the east, from 20 m to 100 m above sea level. Among these ridges there are 20-40m deep depressions filled with soils. This geomorphologic shape continues to the west in to the Sinai Desert, and to North East to Magdal. Among different quaternary soil deposits, the sand dunes are of special interest. They are formed by wind (eolic), and located along the seashore. Their main distribution is in the South and the North. Sand is extensively quarried for construction purposes, e.g. for local use and export to Israel. The quarrying is an encroachment both on the landscape and the geomorphologic history of Gaza.

2.4 Utilities

The IUG has been established a number of research units and centers in different fields. The Environmental and Rural Research Center (ERRC and Environmental and Earth Science Department (EESD) have been established in 1991 as a part of the IUG program to contribute in developing the environmental research studies and to serve the Palestinian community with graduate a highly qualified persons in this vital and very important field that can contribute in solving the environmental problems and developing the country.

The IUG has a good infrastructure (water, power, networks and e- communication……) to carry out the planned activities of the project (See annexes)

2.5 Environmental Impacts

The project originally is oriented mainly to protect the human health and marine environment. The overall impact will be positive in the different environmental issue in local and regional levels.

In addition, all previously reported literature on the use and application of BT as a biocontrol agent for mosquitoes proved that it is safe because it is specific and has no harmful effects on health and environment.

2.6 PROJECT COST ESTIMATE

The following tables summarized the different cost estimates of the project:

1- FINANCIAL SUMMARY:

BUDGET LINE / Amount in $US
1 / Direct Personal costs (Local) / 127600
2 / Travel and subsistence expenses / 27,200
3 / External assistance / 10,000
4 / Equipment / 87,850
5 / Consumables / 19,500
6 / Results reporting / 5,500
7 / Other Program Supporting Costs / 56,000
Total / 333,650

BUDGET LINE 1:

DIRECT PERSONNEL COSTS FOR PROJECT IMPLEMENTATION (LOCAL)

Professional
category / Number
of persons / Monthly
rate / Number
of Months / Total
amount
Project Team leader / 1 / 1200 / 24 / 28,800
Senior technical manager / 1 / 1200 / 24 / 28,800
Technician workers (Graduate Students) / 5 / 400 / 12 / 24,000
Intermediate technical level staff / 3 / 700 / 12 / 25,200
Engineers / 2 / 1000 / 5 / 10,000
Other [Secretary] / 1 / 450 / 24 / 10,800
Total / 127600

BUDGET LINE 2:

TRAVEL & SUBSISTENCE COSTS

Items / amount $US
A. / Travel in the project area, Site Visits, Sampling, ....
- Transportation / 15,000
B. / Mission Outside the project area
- travel 10 trips X 700 $US
- subsistence (40 days X 130 $US ) / 7,000
5,200
Total / 27,200

BUDGET LINE 3: