ENHANCING POISON BAIT ACCEPTANCE AND TRAPPING OF Rattus rattus USING POULTRY EGG COMPONENTS AS BAIT...

CHAPTER – I

INTRODUCTION

Rodents have been identified as the most important

mammalian pests at the global level (Cuong et al 2002). They

cause significant damage to crops at pre- and post-harvest

stages throughout the world (Prakash 1988, Buckle and Smith

1994, Parshad 1999a, Singleton et al 1999, Amusa et al 2005,

Fayenuwo et al 2007, Meerburg and Kijlstra 2008, Singla and

Parshad 2010, Singla and Babbar 2010 and 2012), with yield

losses of 5-15% in most of the countries (Singleton and Petch

1994, Singleton 2003). Damages caused by rodents lead to huge

amount of food shortages (Palis et al 2007). The rodent fauna of

the Indian sub-continent is represented by 46 genera and 128

species (Ellerman 1961, Roonwal 1987). Of these 18 species are

commensal and agricultural pests. Present checklist in India

reports 103 species and 89 subspecies under 46 genera which

belong to 7 families (Pradhan and Talmale 2011). Of these, 20

species have been reported to be of economic importance in

India (Sridhara and Tripathi 2005). Their habitat,

distribution, abundance and economic significance vary in

different crops, seasons and geographical regions of the

country (Rana et al 2006). Despite the development of a wide

variety of rodent control strategies to limit their damage

(Parshad 1999a, Singleton et al 1999, Parshad et al 2006, Weihong

et al 1999), rodent control has not yet become an integral

component of crop production and storage strategies in India.

Analysis of the information available on the damage and

economic losses caused by rodents in various crop fields,

horticulture and forestry, poultry farms, rural and urban

dwellings and storage facilities showed that chronic damage

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ranging from 2 to 15% persisted throughout the country

(Parshad 1999a). There are reports on rodents causing 6-8%

losses in rice crop and 10-12% in wheat crop in India (Santra

and Manna 2008, Chattopadhyay et al 2010). Annual food losses

caused by rodents in India are estimated to be 10-15% of the

total national production (Malhi and Sheikher 1989). Storage

losses to rodents in India alone are 25-30% costing at least

$5 billion annually (Cao et al 2002). In addition, rodents also

destroy food by contaminating it with their urine, faecal

droppings and hair. They are also important vectors or

reservoirs of numerous diseases that infect humans, domestic

animals and other wildlife species (Gratz 1994, Singla et al

2003, 2008a and 2012, Pai et al 2005, Meerburg et al 2009). They

are known to serve as hosts for at least 60 diseases (Hugh-

Jones et al 1995). The need to control commensal rodents is

rarely controversial, as the public has always associated rats

with lethal, disease-causing organisms such as bubonic plague.

While plague has long been absent from some countries,

including India, the potential health threat from commensal

rodents is now focused on other zoonotic diseases, such as

leptospirosis and salmonellosis (Cowan et al 2003).

The reasons for the subsistence of large rodent

populations in most of the storage premises and human

dwellings is the inadequate maintenance of buildings combined

with lack of hygiene, poor handling of food materials leading

to spillage and serious neglect of rodent proofing. The house

rat, Rattus rattus (Linnaeus) is one of the most common commensal

rodent pest worldwide. It often damages, contaminates and

spoils packed food and non-food materials in transit and

storage (Parshad 1999a). It is purely an indoor pest in

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Pakistan (Roberts 1977, Brooks et al 1987, Ahmed et al 1995, Khokhar

et al 1999, Hussain and Iqbal 2002). It is the predominant pest

species infesting and depredating poultry farms in India with

highest annual productivity of 69.59 young/female/year

reported for any Indian rodent species (Sridhara and

Krishnamurthy 1992, Parshad 1999b).

Poultry is one of the important meat industries of India.

Poultry farms provide a most favourable and stable habitat

throughout the year for large populations of R. rattus (Parshad et

al 1987). It causes severe economic losses to poultry

production directly by feeding on poultry feed, contaminating

it with their excrements, damaging eggs, attacking and killing

chicks, causing structural damage to buildings, doors, windows

and feed containers and indirectly by transmitting or carrying

several diseases (Ahmad et al 1984, Chopra and Dhindsa 1987,

Parshad et al 1987, Soni and Rana 1988, Chopra 1992, Sridhara and

Krishnamurthy 1992, Gomez Villafane et al 2001, Hussain et al

2006, Chopra et al 2008a and b). The species acts as a wild

reservoir horizontally transmitting infectious organisms to

other rodent species and arthropod vectors living closer to

anthropized environments, thus leading to inevitable exchange

of pathogens between rodents, animals and humans (Chopra et al

2008b). Hence it is very important to control this species.

A wide range of control measures have been used from time

to time for the control of commensal rodents depending upon

the ecological conditions. Conventional methods of rodent

control all over the world include proofing, trapping and the

use of rodenticides (Parshad 1999b, Weihong et al 1999, Parshad

et al 2006). Trapping rats is an age old method which is

ecologically sound and environment friendly. Some scientists

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have shown that trapping can, under some circumstances, be an

effective method of rodent pest management (Gebauer et al 1992,

Tobin et al 1993, Islam and Karim 1995). The use of appropriate

baits is, however, important when attempting to attract

certain species into traps (Sivaprakasam and Durairaj 1995,

Szocs et al 2004, Fuller et al 2005). Currently, the use of

rodenticides is the major method of rodent pest management.

Acute rodenticides and anticoagulants are the most effective

and widely used groups (Buckle 1994). Both have their own

merits and demerits and are recommended for use against

rodents under different conditions. Out of these two kinds of

rodenticides, acute poisons are more preferred and frequently

applied as people are anxious to see a rapid kill and get rid

of damage caused by them (Sterner et al 1996, Steven 2008).

Rodent control with rodenticide baits in crop lands and

other premises has often been found ineffective in reducing

rodent densities due to several factors (Pank 1976). Although

zinc phosphide has been reported to be an effective acute

rodenticide (Matschke et al 1982, Sterner et al 1996), numerous

researchers have reported bait acceptance problems related to

bitter taste, sub-lethal toxicosis and subsequent conditioned

aversion after rodents have ingested minimal levels of zinc

phosphide bait (Reidinger and Mason 1983, Prakash and Ghosh

1992, Reidinger 1997) resulting in reduced efficacy of

rodenticide baiting (Rzoska 1953, Prakash and Jain 1971,

Berdoy and MacDonald 1991, Sterner et al 1996). In no-choice

tests, zinc phosphide often produces 100% mortality, but in

bi-choice tests that more closely simulate its use in crop

fields, mortality levels can range from 50 to 75%.

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Low bait uptake often occurs in connection with enhanced

neophobia (Berdoy 1994). There is, however, no evidence for a

genetic basis of different levels of neophobia in rat

populations (Macdonald et al 1999). Rats show initial avoidance

towards unfamiliar bait and new bait boxes in a familiar

environment (Barnett 1958). After initial intake of sublethal

dose, rats rapidly learn to avoid eating a poisonous mixture

or particular cereals used as bait base and become bait shy

(Howard and Marsh 1970, Bhardwaj and Khan 1979). Bait-shyness

induced through conditioned taste aversion, can last more than

a year, even when the rodenticides have been removed from the

baits (Howard and Marsh 1970, Owan 1978, Owan et al 1979,

Shepherd and Inglis 1993). Once shy, the rats prefer to remain

hungry than eating an apprehensive food (Sood and Gill 1980,

Prescott et al 1992). Several factors may influence selection of

bait by different species such as taste (Frank 1988),

olfactory sensitivity (Vander Wall et al 2003) and learned

feeding behavior (Galef and Whiskin 2001).

Several researchers have reported the need for an

additive that could be added to rodenticide baits and traps

(Smythe 1976, Marsh 1988, Shafi et al 1990, Parshad and Jindal

1991, Shafi et al 1991, 1992a, b, c and 1993, Koehler et al 1994,

Reidinger 1997, Khan et al 2000, Pervez et al 1999, 2003 and 2005)

for increasing their efficacy. Effective attractants would

allow managers to attract rodents to remote detection devices,

bait stations, or traps. The bait stations, in turn, might

have oral baits containing rodenticides or fertility control

materials (where non-target hazards preclude lethal control).

Most research along these lines, however, has been on food

flavor additives to enhance palatability and the amount of

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time spent feeding on toxic baits (Meehan 1984, Marsh 1988).

Unfortunately, identification of effective rat attractants has

eluded researchers to date. Bullard (1985) reported that rats

were more interested in familiar foods than in a wide array of

odours tested. Additionally, much more effort has been put

into research on rodent repellents than on attractants (Meehan

1984). Bait additives have a long, if not ancient history.

But, till date, no rat attractant has ever been developed. The

best "attractant" so far is just a good bait.

Different workers have suggested the efficacy of

different locally available bait additives against different

rodent species (Kumar and Gangwar 2001, Kamal and Hossain

2003, Pervez et al 2003, Kaur and Parshad 2005, Alam et al 2007,

Witmer et al 2010, Naeem et al 2011, Kok et al 2013, Mushtaq et al

2013). Usually it is recommended that poultry feed may be usedas baiting medium to contain rodent menace in poultry farms

(Bhardwaj 1983). There are some reports on preference for

poison bait containing egg shells and egg contents by rodents

in rice and wheat crop fields, respectively (Khan et al 2000,

Pervez et al 2005). Usually it is recommended that poultry feed

may be used as baiting medium for preparation of rodenticide

baits to curtail rodent menace in poultry farms (Bhardwaj

1983). These additives if added in zinc phosphide bait may

benefit the farming community in increasing its acceptance as

it still remains one of the most widely used rodenticides in

the subcontinent.

Since R. rattus is the predominant rodent pest species found

in poultry farms and feeding on poultry eggs, the bait

containing poultry egg components may be more acceptable to

it. Present study was therefore proposed to formulate a cereal

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based bait package for R. rattus using poultry egg components such

as egg shell and egg albumin for enhancing poison bait

acceptance and trapping of R. rattus with following objectives:

i) To study the acceptance of different concentrations of egg

shell and egg albumin by R. rattus in laboratory experiments.

ii) To study the acceptance of best concentration of two

bait additives by R. rattus under food scale consumption

monitor.

iii) To determine the effect of best concentration of best

bait additive in enhancing poison bait acceptance and

trapping by R. rattus in poultry farms.

7

CHAPTER – II

REVIEW OF

LITERATURE

Rodents inflict incalculable losses to standing crops,

harvested crops in threshing floors and to stored food grains

and other commodities. Their damaging propensities in

rangelands, afforested lands, fruit orchards and plantation

crops and poultry farms are well known (Parshad 1999b). No

country in the world is free from quantitative and qualitative

losses inflicted by rodents. But the losses are more

pronounced in the tropical countries where rodents play a

significant role in retarding agricultural production through

damage to field crops (Jackson 1977, Parshad 1999a, Singla and

Parshad 2010, Singla and Babbar 2010, Borah and Bora 2012,

Gogoi and Borah 2013). The damage caused by field rodents maysometimes be estimated roughly for local districts where a

measurable proportion of various crops is destroyed by the

rodents. Commensal rodent damage is, however, difficult to

assess primarily because so many different items can be

involved and rodents can invade almost any type of structure

(Lund 1994). In food stores and warehouses, rodents can give

great problems, not only by consuming or fouling a substantial

part of the food, but also because they destroy sacks, bags,

boxes and other packaging material. In the slum areas of large

towns and certain villages in developing countries, there is a

constant fight between humans and rodents when hungry rats try

to gnaw fingers and toes of elderly and helpless people during

the night or babies when not looked after (Lund 1994). Rodent

consumption of stored food grains and damage to storage

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structures and containers, and indirect losses caused by

spillage, spoilage, or contamination by hair or faecal

droppings that results in condemnation or rejections of

shipments by the importing country are important economic and

public health problems worldwide (Jackson 1977, Brooks and

LaVoie 1990, Conover et al 1995).

Rodents are very intelligent animals with a strong

smelling and tasting sense, which prevents them from instant

ingestion of poisons applied directly. The rodenticides are

therefore applied with different bait materials. But again

their behavioral characteristic termed as neophobia defends

and restricts them to immediately switch on to poison baits.

Such behavioral phenomena render the rodents control very

tricky, because once shy, the rats prefer to remain hungry

than eating an apprehensive food (Canby 1977, Riley and Clarke

1977, Owan 1978, Owan et al 1979, Sood and Gill 1980, Prescott et

al 1992). Many workers have tried to find out the most

attractive bait materials (Shumake 1978). Because of the large

space or diastema behind their incisors, rodents can use

incisors to investigate or nibble unfamiliar materials without

actually taking them inside their mouths. Rodents are

omnivorous, exhibiting choices and preferences in their diet,

but often selecting the most abundant, palatable food

available. They readily learn to reject or avoid unpalatable

foods or those containing toxins, which presents a problem for

the development of bait materials for effective delivery of

rodenticides.

2.1 Commensal rodents

Rodent pest species as commensal, literally means ‘eating

at the same table’. Pest rodents eat man’s foods, damage his

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possessions and use his transport to distribute all over the

globe. Rodent species of the genus, Rattus have probably been

responsible for more human sufferings than any other group of

vertebrates, not only through their destructive impact on food

crops, but also through their role in the transmission of

fatal or debilitating diseases. However, the genus also

contains species of immense scientific and clinical

significance, such as the familiar Norway or laboratory rat

(Rattus norvegicus), as well as a host of lesser known but

presumably beneficial species that inhabit a wide range of

natural habitats from mangrove forests to subalpine

grasslands. The genus Rattus, with a total of 61 currently

recognised species, is the only single largest mammalian genus

of all which is most complex and least well understood. R.

rattus and R. norvegicus, both are the best-known commensal species

having global distribution (Aplin et al 2003). The house mouse

(Mus musculus) is also widely distributed and together with the

house rat and Norway rat, are known as commensal rodents

because of their close association with human habitation.

Since most rodent species involved in stored product damage

are nocturnal, heavy infestations may persist unnoticed

without careful inspection of stores or premises (Jackson

1990).

2.1.1 The house rat, Rattus rattus

The house rat, R. rattus (Rodentia: Muridae), is one of the

most commonly encountered and economically important commensal

rodents. It is nocturnal and prefers to forage for food above

ground in elevated areas indoors and outdoors. It is an agile

climber and travels through trees and along vines, wires,

rafters, and rooftops. It often uses trees and utility lines

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to reach food and to enter buildings, but can also be found

foraging in dense ground cover. It can swim and may use sewer

systems to disperse to new areas. Outdoors, it can travel

several hundred feet in a single night to find its survival

resources. It prefers to nest in secluded areas above ground

in such places as attics, soffits, overhead garage storage, in

the vine cover of fences or buildings, and in wood piles or

other stored materials where harborage can be found (Askham

and Leonard 1987). It favours dense non-deciduous trees or

trees with hollow cavities and the crowns of palm trees,

especially when old fronds are not removed. It sometimes

burrows in the ground especially in hot, dry environments. In

these areas, it may use trees, materials stored on the ground,

concrete slabs and sidewalks to support shallow burrows. Roof

rat signs include smudge marks on surfaces from oil and dirt

rubbing off their fur as they travel. Sounds in the attic are

often the first indication of the presence of house rats in a

residence. At night when the house is quiet, the rats may be

heard scurrying about (Sullivan 2002).

The house rats have a high reproductive potential and may

breed year-round in warmer areas (Chattopadhyay et al 2010).

They can reproduce when they are 3 to 5 months old. Females

produce 5 to 8 pups per litter with average 4 to 5 litters per

year. The number of litters depends on the area and varies

with nearness to the limit of their climatic range,

availability of nutritious food, density of the local rat

population and the age of the rat (Marsh 1994). The gestation

period is between 21 and 29 days. The young are born in a

nest. At birth they are hairless and their eyes are closed.

The young ones in the litter develop rapidly, growing hair

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within a week (Sullivan 2002). Between 9 and 14 days, their

eyes open and they begin to explore for food and move about

near their nest. In the third week, they begin to take solid

food.

The house rat has been implicated in the transmission of

a number of diseases to humans and domestic animals (Gratz

1997). Examples include murine typhus (rickettsial disease),

various spirochetal diseases (lyme disease, leptospirosis) and

some protozoal diseases (leishmaniasis, toxoplasmosis). Of

these, leptospirosis is of special concern as an emergent

infectious disease, with a recent upsurge in the rate of

diagnosis of this previously ‘hidden’ disease (Singleton and

Petch 1994). Commensal house rat populations have been found to

carry heavy leptospira infections in widespread regions of

Africa, Asia, Europe and North America (Gratz 1997). These are

also suspected in the transfer of ectoparasites from one place

to another. The role of R. rattus as an agent of disease is most

spectacularly illustrated through the example of the great

plague pandemics. House rats or possibly just their fleas,

harbouring in cloth and other trade goods, first carried the

plague organism (Yersinia pestis) from Central Asia to the Middle

East and Europe in the 5th century AD. Shipping routes

provided even more effective means of transporting plague

bearing rats around the world, leading to the infamous Black

Death of the 14th century. Plague remains endemic in Central

and Southeast Asia, many parts of Africa and South America and

across much of the United States of America. In each area,

rodents serve as the primary or enzootic hosts (Biggins and

Kosoy 2001), with the pathogenic agent maintaining a sensitive

balance between infection rate and death rate in order to

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persist indefinitely in the wild population. During the latest

resurgence of plague in India in 1994, about 4000 persons

suffered from infection and about 100 persons died and the

resulting panic led to tremendous loss of agricultural and

industrial production (Ramalingaswami 1994).

In Hawaii, North and South America and in Africa, house

rats are seen damaging ripening sugarcane crops (Taylor 1984,

Fiedler 1988, Prakash and Mathur 1988, Wood 1994). Coconuts

grown commercially in many tropical areas are subject to

damage particularly by R. rattus and R. tanezumi (Tobin et al 1990).

These species climb palms of all ages, primarily to feed on

developing nuts, which then fall prematurely on the ground

(Fiedler et al 1982, Wood 1994). The proportion of nuts that drop

prematurely due to rat damage can be quite high. However,

impacts on yield may not be proportional to the number of

developing coconuts that fall to the ground (Williams 1974,

Reidinger and Libay 1981, Fiedler et al 1982). Trees in some

areas may compensate for early damage by increasing the size

and weight of remaining nuts (Tobin et al 1997). The economic

impact of this damage is thus not clear (Tobin and Richmond

1993). In situations where rats feed on coconut flowers or

damage very small nuts, yield losses may be underestimated by

counts of fallen, maturing nuts. Macadamia orchards in Hawaii

and Australia sustain extensive damage from R. rattus (Tobin 1992,

White et al 1997). Five to ten percent of developing nuts are

damaged by rats in some Hawaiian orchards. The rats feed on

the fruit and vegetative portions of many landscape and garden

plants including the bark of trees. Their feeding and gnawing

may completely girdle young trees.

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The house rat poses a serious threat to poultry

operations by feeding on poultry feed, contaminating it with

their excrements, damaging eggs, attacking and killing chicks,

causing structural damage to buildings, doors, windows, feed

containers and transmitting or carrying several diseases

(Ahmad et al 1984, Khatri and Veda 1984, Parshad et al 1987, Soni

and Rana 1988, Gora et al 1995, Gomez Villafane et al 2001,

Hussain et al 2006, Chopra et al 2008a and b). Even poultry

buildings with fibre glass or polystyrene insulated walls are

vulnerable to damage (Turner 1986). In addition, they also

cause general nuisance for birds in the poultry house due to

their noise and movements. The birds may be frightened, which

results in poor performance. However, due to financial

limitations many farmers build poor quality sheds inside the

crop fields and close to villages which naturally attracts

both field and commensal rodents leading to considerable

financial losses (Chopra and Sabharwal 1992, Gora et al 1995,

Munjal 2000, Chopra et al 2008a). A high carrying capacity of

poultry farms for rodents is evident from the reports of 292

rodents/3600 sq. ft floor area (Malhi et al 1991) and 72

rodents/100m2 floor area (Ahmad et al 1992). Damage to poultry

feed is a major cause of economic loss as feed accounts for

50–75% of the operational cost of a poultry farm. By causing

frequent structural damage to wooden doors, windows and

electric cables (by gnawing) and to floor and foundations (by

burrowing) the rats increase the maintenance costs

of the building.

2.2 Management methods

Different methods of commensal rodent management include

environmental sanitation, habitat manipulation, rodent-

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proofing, use of chemosterilants, ultrasonic and

electromagnetic devices, traps, repellents and rodenticides

(Greaves and Rowe 1969, Kassa and Jackson 1984, Buckle 1994,

Smith 1994, Prakash et al 2003, Tobin and Fall 2006, Singla and

Parshad 2000 and 2009, Singla and Mittal 2012, Singla and Garg

2013, Singla et al 2008b and 2013). There has been heavy

reliance on rodenticides to control rat populations, although

other methods such as traps and exclusion are also used (Timm

1994). Traditional baiting or trapping on the ground or floor

may intercept very few house rats unless bait and/or traps are

placed at the very points that rats traverse from and to a

food resource. House rats have a strong tendency to avoid new

objects in their environment and this neophobia can influence

control efforts, for it may take several days before they will

approach a bait station or trap. Neophobia is more pronounced

in house rats than in Norway rats (Marsh 1994). The use of

rodenticides, in the form of poison bait is inexpensive and

effective. The success of this programme is, however, based on

the type of poisons and their formulations (Prakash and Mathur

1992). Baiting technique if appropriately applied is the most

reliable strategy to control rodent pests. Behavior modifying

components may play a significant role in developing the most

attractive baits. The criteria for food selection in rats are

complex and may depend upon many factors including

palatability (Young 1946). Taste of food plays a significant

role in food preference. According to Marsh (1986) taste often

supersedes any earlier influence of odour in food selection to

a degree that is not paralleled in humans.

2.2.1 Attractancts for rodenticide baits

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Rodent control with poison baits in crop lands is often

ineffective in reducing rodent densities due to several

factors. Poor bait acceptance, sub-lethal dosing and dietary

preferences can reduce the efficacy of rodenticide baiting

(Berdoy and MacDonald 1991). Although zinc phosphide has been

reported to be an effective acute rodenticide (Matschke et al

1982, Sterner et al 1996), numerous researchers have reported

bait acceptance problems related to bitter taste, sub-lethal

toxicosis and subsequent conditioned aversion after rodents

have ingested minimal levels of zinc phosphide bait (Reidinger

and Mason 1983, Prakash and Ghosh 1992, Reidinger 1997). Bait-

shyness, induced through conditioned taste aversion, can last

more than a year, even when the zinc phosphide has been

removed from the baits (Shepherd and Inglis 1993). It has been

established that field as well as commensal rodents develop

poison aversion and bait shyness after a single exposure to

the widely used rodenticide, zinc phosphide (Prakash and Jain

1971, Prakash et al 1975, Bhardwaj and Khan 1979, Saxena and

Mathur 1995). The persistence of shyness in rodents which are

not pre-baited prior to poison baiting is of a higher

magnitude than in those which are exposed to the toxicant

after prebaiting (Bhardwaj and Prakash 1982). The development

of poison aversion and bait-shyness among rodents of economic

importance makes the control of any residual population

surviving a zinc phosphide baiting operation a difficult task,

since bait carrying the same poison on the second day is not

accepted by them (Bhardwaj and Prakash 1979). Sampling of food

is well known in R. rattus (Barnett 1966), R. norvegicus (Cowan 1977,

Barnett et al 1978) and B. bengalensis (Parshad and Jindal 1991).

Even in natural habitats, this sampling behaviour has survival

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value as it enables the rats not only in finding new sources

of food but also in avoiding toxic baits (Barnett 1966,

Siddiqui and Khan 1982, Chopra et al 1984 and 2009).

Several researchers have noted the need for an attractant

odour that could be added to zinc phosphide baits to

successfully compete with the alternate field crop foods

(Koehler et al 1994, Reidinger 1997, Khan et al 2000). A more

rapid consumption rate for rodenticide baits may also lead to

a lessened need to leave baits exposed to non-target species

over extended time periods (Watkins et al 1999).

2.2.1.1 Pheromones as attractants

Pheromones are known to play an important role in the

social behaviour of mammals like attraction to opposite sex,

aggregation, parental care, individual identification,

territorial marking etc. The major source of chemical cues

involved for pheromonal communication are the secretions of

the specialized scent glands, urine and feces. In rats, the

preputial glands and cheek glands are reported to be important

as they stimulate a variety of social functions. Preputial

glands, in particular, appear to play an important role in

production of olfactory substances which attract the opposite

partner. When faced with a choice among several novel foods,

naïve (observer) rats choose novel foods that have previously

been ingested by conspecifics (demonstrators) with whom they

have interacted (Strupp and Levitsky 1984). This socially

mediated transfer of food preference has been observed even

when demonstrators are anesthetized and wire-mesh barriers are

placed between demonstrator and observer (Galef and Wigmor

1983). Such findings and results of other experiments (Galef et

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al 1985) suggest that diet preference is mediated in part by the

volatile chemical cues.

Researchers have shown that rodents will respond to the

biologically-derived odors of conspecifics, but the responses

are variable depending on the age, sex, social dominance and

breeding condition of the animal (Salmon and Marsh 1989,

Drickamer 1997). Consequently, in terms of management tools,

we cannot expect rodents to respond reflexively and

consistently to pheromones as do some insects (Howard 1988).

One of the few odours that some researchers have found to be

attractive to rodents is carbon disulphide (Galef et al 1988,

Shumake et al 2002), however, other researchers did not find

this material to be particularly attractive (Koehler et al

1994). Carbon disulphide present in the rodent breath is

responsible for inducing diet preferences in rodents (Galef et

al 1988). The addition of carbon disulphide to a bait has also

been shown to increase bait consumption and time spent at that

site (Bean et al 1988). Carbon disulphide is therefore a

biologically meaningful odour to rodents that increases

attractiveness of foods. However, despite the publication of

these results, carbon disulphide was never used in commercial

baits. This may in part be due to its volatility and/or

toxicity. Although environmental marking by animals generally

relays social, sexual, or reproductive status, it has also

been shown that scent marking by Norway rats may play an

additional function in communicating food preferences.

Excretory deposits that surround food sites render those

sites, and the food, more attractive to Norway rats than

unmarked sites (Laland and Plotkin 1991). Further manipulation

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of this behaviour at a bait station may not lure a rat to the

site but, should assist in bait uptake.

Conspecific urine odour is reported to improve poison

bait acceptance by reducing neophobia and masking poison

aversion and bait shyness behaviour in Bandicota bengalensis,

Meriones hurrianae and Tatera indica (Kumari and Prakash 1980 and 1988,

Kaur and Parshad 2006a). Scent gland secretions (Selvaraj and

Archunan 2002) and urine (Soni and Prakash 1987) have been

reported to overcome poison bait shyness to some extent in

laboratory rats and soft furred field rats, respectively.

The sex pheromone which is found in female rodent urine

or vaginal discharge could be an important substance for

attracting males to traps or to stations with toxic bait or

chemosterilants. An attempt to evaluate this potential

possibility has been made by Field (1971) who reported

increased acceptability of chemosterilant bait by rats on

treatment with estrous urine. However, more research is needed

to quantify the attractant capacity of these pheromones, and

to isolate and identify the chemical components that are

responsible for this presumed attraction. Odour plays a

considerable part in sexual activity and sex lures have been

used successfully in insect control. Proprietary rodent baits

are available “with added sex attractant" but the sex for

which the attractant is designed is not specified. The use of

sex attractants has considerable appeal to the public but

their use in rodent control has yet to be substantiated (Bull

1972).

2.2.1.2 Poultry egg components as bait additives

Pervez et al (2000) reported 3% egg yolk to be the

preferred bait additive over the yeast powder, minced meat and

19

`

fish meal, for controlling short tailed mole-rat, Nesokia indica

in laboratory studies. Shafi et al (1990) reported the

preference of egg yolk over egg shell powder, against the wild

caught R. rattus in Pakistan, while Pervez et al (1999) suggested

preference of egg shell powder over egg yolk by field rodents

(B. bengalensis, N. indica, Millardia meltada and Mus spp.). The egg, when

used as additive (2%), allowed a higher consumption of poison

bait for a successful control (Pervez et al 2005) and 3% egg

additive could mask the undesirable characters of the bait

material, rendering it palatable to commensal R. rattus and field

rodents (B. bengalensis and N. indica) (Pervez 2007). Abbas (2003),

however, reported preference of rats towards peanut butter

than egg shell powder as additives to bait in B. bengalensis in

wheat crop.

Shafi et al (1990) recommended that 2% egg yolk as additive

in poison bait can be useful in enhancing bait acceptance by R.

rattus in an effort to reduce its infestation in various

situations. It increased acceptability by 72.7 and 67.9% of

the bait containing brodifacoum and bromadiolone, respectively

against reference poison bait. Khan et al (2000) reported

preference for poison bait containing egg shells by rodents in

rice crop fields. Two second generation anticoagulants i.e.

brodifacoum and bromadiolone and one acute rodenticide i.e. zinc

phosphide each mixed with whole egg were tested in three

separate blocks of wheat crop (Pervez et al 2005). High intake

of egg mixed zinc phosphide bait by rodents was reported by

Prakash (1976). The improved formulation of zinc phosphide may

benefit the farming community as it still remains one of the

most widely used rodenticides in the subcontinent. Shafi et al

(1992b) reported highest intake of brodifacoum mixed with egg

20

`

shell by field rodents of wheat, over other additive baits.

Similar pattern of bait intake was recorded in bromadiolone

and zinc phosphide baits.

Khan et al (2000) conducted field trials in rice crop to

control rodent populations by improving poison bait acceptance

using the additives viz., minced meat, egg-yolk, egg-shell and

yeast. Poison baits, brodifacoum (0.005%), bromadiolone

(0.005%) and zinc phosphide (2%) were employed as

rodenticides. Egg-shell ranked first in the preference over

the other additives and non-additive poison baits. Mixing of

additive with brodifacoum reduced the population of M. musculus

to 92.4%, with bromadiolone to 89.7% and with zinc phosphide

to 94.7%. Similarly, for B. bengalensis, a reduction of 91.5% with

brodifacoum, 92.0% with bromadiolone and 94.7% with zinc

phosphide was recorded, while for both N. indica and M. meltada,

the respective decrease of 87.5, 90.5, 94.2, and 88.7, 85.7

and 94.7% was reported. Two baitings, one at flowering and

other at maturity stage of the crop reduced damage to 94.0%

with brodifacoum, 92.0% with bromadiolone and 91.5% with zinc

phosphide. It was concluded that the two baitings, one at the

flowering stage, and the other at maturity stage may be

employed to obtain a robust production.

It was suggested that the texture of the egg shell marks

an attractive taste for the rodents. For the zinc phosphide,

more preference for the egg shell bait than the non-additive

bait was significant as fundamentally, zinc phosphide causes an

immense "bait-shyness" and poison aversion (Prakash 1976). It

was hypothesized that an improved formulation using zinc

phosphide and egg shell may prove beneficial to the farmers as

it is one of the widely employed acute rodenticide against the

21

`

control of rodent species. However, with lack of expertise, its

grievous consequences cannot be done away with readily.

Addition of egg shell and fish meal did not show a significant

increase in relative consumption for the cereal base by B.

bengalensis (Naeem et al 2011). Rather they observed an inverse

trend indicating decreased preference for plain millet based

bait when mixed with egg shell and fish meal. Shafi et al (1993)

observed high preference toward minced meat by B. bengalensis than

egg shell and egg yolk mixed bait.

2.2.1.3 Other bait additives as attractants

Meehan (1984) cautioned that it is impossible to say

which particular foodstuff will be preferred by individual

rats or even whole populations. There is no such thing as

‘universally acceptable bait’. Rowe (1973) made a similar

comment about mice. These statements have been confirmed by

studies of consumption of foods by wild rats (Clark 1982).

Lund (2008) gave a general summary of the ideal

characteristics of rodent baits and additives. No magical

additives that made baits irresistible were identified. Marsh

(1988) summarised that sugars and vegetable oils and animal

fats are the most universally effective additives for cereal

baits to improve acceptance and palatability. Flavour

additives to bait have often decreased rather than increased

consumption.

A variety of methods were used to control rodent

populations and to reduce their damage (Witmer et al 1995).

Effective odour attractants would allow managers to attract

rodents to remote detection devices, bait stations, or traps.

The bait stations, in turn, might have oral baits containing

rodenticides or fertility control materials. Witmer et al (2008)

22

`

tested 3 odor attractants i.e. almond, lemon and ginger, that

appeared to have potential for effectively attracting Norway

rats but, none of the 3 potential attractants were found

effective. Wines (2004) examined the efficacy of liquid

concentrated banana extract as a potential attractant for

trained Gambian rats.

Bullard and Shumake (1977) used the responses of

Philippine rice field rats (R. rattus mindanensis) to show that

intensifying the flavour cues of familiar or favoured foods

could be a useful way of enhancing bait intake. Such

attractants help to keep rodents feeding longer at food

sources. They may be effective because they mask the taste of

a rodenticide and/or they are palatable in their own right

(Meehan 1984). Sugar is a well-known effective additive for

rats and mice, while bitter flavours tend to be rejected (Rowe

1961, Howard et al 1972, Shimizu et al 1980, Marsh 1988, Yamaguchi

1995). Norway rats in trials preferred salty and sweet tastes

over sour and bitter ones (Karasawa and Muto 1978, Kolody et al

1993). Singla et al (2010) studies the efficacy of sodium

chloride in improving acceptance of bait by rodents. The most

attractive concentrations of sugars have also been assessed in

various trials (Collier and Bolles 1968). When given the

choice between sugar on its own and flour, Norway rats

consumed only small amounts of the sugar (Barnett 1956).

Inverted sugars like maltose, dextrose, fructose and levulose

also are acceptable to rodents (Smythe 1976). Of the

artificial sweeteners, saccharin was favoured by laboratory

rats over cyclamate, while wild Norway and house rats

preferred glucose to saccharin (Wagner 1971a and b). Sugars

also help preserve the baits, potentially increasing shelf and

23

`

field life, but also increase bait palatability to

invertebrates and reptiles. Babbar et al (2009) found higher

acceptance of zinc phosphide bait containing 2% sugar by R.

rattus. Mice in a study exhibited a preference for monosodium

glutamate and sodium chloride (common salt), but more so for

sugars (Yamaguchi 1995). Salt and monosodium glutamate are

variable in their effect, and were disliked at concentrations

above 0.5% (Ohara and Naim 1977, Marsh 1986 and 1988, Kolody et

al 1993). Female rats consumed more salt than male rats (Flynn

et al 1993).

Oils can be effective bait additives. Meehan (1984) found

that the higher was the level of oil in bait, the more readily

it was taken. Mice have exhibited a preference for high-fat

foods (Imiazumi et al 2001) and rats have an appetite for

dietary fats and oils (Elizalde and Sclafani 1990, Ramirez

1993). Glycerine, corn oil, arachis oil and mineral oil were

more palatable to mice than olive, linseed or cod-liver oils

(Rowe et al 1974). Groundnut oil probably has a neutral flavour

to rodents (neither attractive nor repellent), but may act to

mask the flavour of cereals to which rats have developed bait

shyness (Bhardwaj and Khan 1979). In conjunction with gur,

groundnut oil increased bait consumption by mice more than

just the gur alone (Pathak and Saxena 1995). Olive oil

enhanced the acceptance of baits to male house rats in no-

choice tests, while soybean oil, sunflower oil, mustard oil,

groundnut oil and coconut oils were all preferred over plain

bait in choice tests, and groundnut oil was preferred over

coconut and mustard oils (Ahmad et al 1994). However, in the

same study, female rats preferred coconut, mustard and to some

extent, sunflower and soybean oil in mixed diets. In a study

24

`

in the USA, coconut, peanut and corn oils were most preferred

by Norway rats, whilst corn oil was preferred over peanut oil

by house rats (Meehan 1984).

In Australia, fishmeal was a successful additive to bait

for mice (Robards and Saunders 1998, Jacob et al 2003). These

authors listed a range of other additives such as chocolate,

cheese, aniseed, peanut oil and honey, that made no difference

to consumption. By contrast, chocolate is used elsewhere as an

effective trap attractant for mice to such an extent that UK

researchers have developed a chocolate mouse trap (Singh

2003). The addition of 2% onion pulp improved consumption of

bajra flour by mice in India (Saxena et al 1995). Toasted

coconut has been used successfully as bait in traps in

Rarotonga (Robertson et al 1998). Tabuchi et al (1991) listed

black pepper, milk and coffee as highly preferred food-related

odours, while, nut, peppermint, plum, orange and cheese also

elicited a bar-pressing response from rats. Raw fish and beef,

dried dog food, coconut oil, fresh or dried blood, chicken

offal, cinnamaldehyde, raspberry, aniseed and other commercial

products have all been attributed with attractive properties

for Norway rats (Bull 1972). McFadden (1984) listed aniseed,

banana, coconut, clove, eucalyptus and vanilla as acceptable

lures for Kiore. Schisla et al (1970) described the success of

‘Dexide’ - a carbohydrate with flavour material that increased

the consumption and efficacy of warfarin to mice, house rats

and Norway rats. Witmer et al (2008) reported almond, ginger and

lemon extract as potential attractants against Norway rats,

but they were not effective under field conditions. Kamal and

Hossain (2003) reported paddy grain as the most preferred bait

by rats in rice field, while dry fish and coconut was the

25

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second and third preferred bait, respectively. The comparison

of average daily intake of rats between foods with and without

additives (2% sugar and 2% groundnut oil) revealed that the

rats showed significant preference for food items with

additives over their plain alternatives in most of the cases.

This preference for foods with additives was maintained over

different days of exposure. Taste of food has already been

reported to influence feeding response of R. rattus both in

laboratory conditions and poultry farms (Khan 1974, Prakash et

al 1980, Munjal 2000).

The preference of rats for a particular food depends more

upon its flavour than its nutritional value (Naganuma et al

1973). Additives like sugar and vegetable oils at 1 to 3%

concentration increase the food intake of rodents as they make

foods more acceptable and palatable to rats (Brooks and LaVoie

1990, Malhi and Kaur 1995). Moreover, selection of sweet food

has also an adaptive value as in natural food. Bitter taste is

associated with occurrence of toxic alkaloids or poisonous

foods while sweetness indicates presence of starch and

carbohydrates and the latter form the major component of diet

of rodents (Barnett and Prakash 1975, Rana et al 1992). Based on

overall mean daily food consumption, different foods were

eaten by R. rattus in the preference order of cracked maize >

cracked wheat > wheat flour > poultry feed > fish meal. Maize

is a major constituent of the poultry feed and (Bhardwaj 1983)

reported high preference for cracked maize than poultry feed

by R. rattus. This contradicts previous report that poultry feed

may be used as bait in poultry farms. Interestingly, cereals

and pulses have been recorded with more consumption than that

of poultry feed in Punjab and Rajasthan (Parshad et al 1987 and

26

`

1991, Mathur et al 1992). Thus, high preference of rats for

cracked maize than poultry feed appeared to relate to its

texture and taste qualities.

Naeem et al (2011) reported that dietary preferences of B.

bengalensis depend upon many factors including particle size,

palatability, taste, flavor and nutritious value of bait base

and the taste enhancers. Experiments performed to evaluate

inclination of the rodents towards cereal bases revealed that

millet was the most preferred over the other cereals. The

cereal bait bases preferred by the B. bengalensis were in the

order of Millet > wheat > maize > rice. The use of 2% butter

oil, however, greatly enhanced intake of cracked millet.

Jackson (1965) revealed that texture, odour and taste play a

significant role in selection of a particular bait base.

Preference for millet as compared to other cereals is also

reported by Parshad and Jindal (1991) who enunciated that B.

bengalensis prefers soft and small sized grains. Watson and Perry

(1954) found that millet was preferred by N. indica due to its

small size. Similar inferences were also drawn by Prakash et al

(1970) and Kumar and Khan (1978). Shafi (1991) argued that

particle size of grain plays an important role in enhancing

bait acceptability. Use of poison baits is still the most

reliable strategy for controlling field as well as commensal

rodents. However, baiting techniques should be modified

according to the psychological characteristics like neophobia,

bait shyness as well as feeding behaviour, exploratory

behaviour, transporting, hoarding and territoriality behaviour

of the target species (Lund 2008).

2.2.2 Bait attractants for traps

27

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Trapping rodents in fields and premises is a common old

practice (Fitzwater and Prakash 1989). Two basic types of

traps are being used, the snap or kill trap and the live trap.

Among the snap traps, the Tanjore bow trap, a low cost bamboo

trap generally used by professional trappers in rice fields

(Neelanarayanan et al 1995); the wooden snap trap, a locally

fabricated trap using timber splinters (Srihari and

Chakravarthy 1992); the urang or arrow trap (Prakash and Mathur

1987) and the break-back spring loaded snap traps with wooden

or jawed iron base (Prakash and Mathur 1987) have been used

traditionally. Among these the break-back snap trap is most

popular. Glue boards are also effective for trapping indoor

rodents (Srivastava and Srivastava 1985). However, these have

not become popular so far because of the cost, hygiene

problems and inhumane method of killing.

Trapping efficiency is dependent on trap type chosen

(Wiener and Smith 1972), type of bait and pre-baiting method

(Chitty and Kempson 1949), social interaction between the

individuals (Ylenon et al 1990) and on effects of recent

trapping efforts. The later includes the effect of odour of

previously trapped conspecifics (Krebs and Boonstra 1984),

competitors or predators (Stodddart 1982) and the previous

experience of being caught in a trap (Tanaka 1963). Alam et al

(2007) evaluated six different baits for trapping rodents in

the poultry farms. Among them the preferred order was dry

fish, coconut meat, potato, bread, soap and wax. Hasanuzzaman

et al (2009) evaluated the effectiveness of bread, potato,

coconut meat, dry fish and paddy grains in trapping B. bengalensis

at different stages of wheat crop. Witmer et al (2010) tested 15

attractants for use in traps for capturing or detecting

28

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Gambian rats. They found conspecific scents from faeces and

urine as the best attractants for single and paired Gambian

rats. They suggested that peanut butter, anise, ginger and

fatty acid scents can also be used as attractants in traps.

Kaur and Parshad (2006b) reported the efficacy of conspecific

urine in enhancing trapping of rodents. Kok et al (2013) used

different baits in traps for surveying small mammals at high

altitude in South Africa and concluded that the most suitable

bait is peanut butter and oats.

CHAPTER – III

MATERIAL AND METHODS

The present study was carried out at Animal House

Laboratory and Rodent Research Laboratory, Department of

Zoology, Punjab Agricultural University, Ludhiana and poultry

farms situated in the campus, Guru Angad Dev Veterinary and

Animal Science University (GADVASU), Ludhiana and village

Ghutani Kalan, District Ludhiana. Approval of the

Institutional Animal Ethics Committee was obtained for the use

of animals.

3.1 Laboratory experiments

3.1.1 Collection and maintenance of animals

For present studies, the house rat, R. rattus of both sexes

were trapped with the help of single catch and multi catch rat

traps from store houses and poultry farms in and around

Ludhiana. In laboratory, rats were acclimatized individually

in cages of size 36 x 23 x 23cm (Plate 1a) for 10-15 days

before the commencement of experiment with food and water

provided ad libitum. Food consisted of preferred cereal based

29

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bait containing loose mixture of cracked wheat, powdered sugar

and groundnut oil (WSO bait) in ratio 96: 2: 2. Proper

hygienic conditions were maintained during the period of

experimentation.

3.1.2 Bait additives used

During present studies, poultry egg components such as

egg shell powder (ESP), egg albumin (EA) and crushed egg

shells (CES) were used as bait additives.

3.1.3 Experimental design of bi-choice feeding tests

During experimentation, mature and healthy rats of both

sexes were weighed and divided into four groups of ten rats

each (5 of each sex in each group). In first experiment, rats

of groups I, II and III were exposed to WSO bait containing 2,

5 and 10% ESP, respectively as additive in bi-choice with WSO

bait for 5 days. Rats of group IV kept as untreated control

were fed on WSO bait without additive. There was no

significant difference in average body weight of rats in

treated and untreated groups. Bait was kept in bowls and water

was provided ad libitum. The position of bowls was changed daily

to avoid any difference in consumption due to site preference.

In second experiment, rats of groups I, II and III were

exposed to WSO bait containing 2, 5 and 10% EA as bait

additive, respectively, whereas in experiment third, similar

groups of rats were exposed to WSO bait containing 2, 5 and

10% CES as bait additive, respectively. In all the three

experiments, rats of group IV were kept as untreated control

and were fed on WSO bait without additive. Before and after

the treatment period, all the rats were fed on WSO bait.

30

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3.1.3.1 Bait acceptance

In all the three experiments, bait consumption was

recorded after every 24 h and every time, bait was replaced to

the original 30g. Before weighing, the bait of all the treated

and untreated rats was cleared of faecal pellets and dried if

needed. Mean daily consumption of food (g/100g bw) was

calculated separately for each group of rats during pre-

treatment, treatment and post-treatment periods. The percent

acceptance of treated bait over WSO bait consumed during

treatment period was determined as per the formula given

below:

Percentacceptance =

Consumption of WSO bait with additiveduring treatment period x

100Total bait consumed during treatment period

3.1.4 Experimental design of no-choice feeding tests

Four rats of each sex were exposed to WSO bait containing

bait additives in no-choice feeding test under Food Scale

Consumption Monitor (FSCM) (Columbus Instruments, USA) (Plate

1b). Each rat was kept in a cage provided with a feeding bowl

and a feeding sensor below. In first experiment, four female

rats were first exposed to WSO bait for four days. Every day

rats were exposed to bait for two hours. Before exposure to

baits, rats were kept without food overnight. Subsequently,

these rats were exposed to WSO bait containing 2%

concentration each of ESP, EA and CES. WSO bait containing

each type of bait additive was exposed to rats for four days.

Behaviour of rats was recorded in the form of food consumption

(g/kg bw), total number of feeding bouts made and total time

spent (s) in bouting in 2 hr a day by means of software of

31

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FSCM loaded in the computer. Similar experiment was carried

out on male rats also.

3.2 Experiments in poultry farms

3.2.1 Bait additives in rodenticide baits

Three blocks (I, II and III) of poultry farm, with each

block further consisting of four replicated sheds were

selected at village Ghutani Kalan, District Ludhiana. One

block (IV) consisting of four sheds selected at the campus,

GADVASU, Ludhiana was kept as untreated control. Before

treatment, pre-census bait consumption was recorded from all

the sheds of four blocks by keeping WSO bait @ 500g/shed in

two rows of ten bait points each to record level of rodent

activity before treatment. After pre-census bait consumption,

sheds of blocks I, II and III were treated with 2% zinc

phosphide bait prepared in WSO without any bait additive, WSO

containing 2% egg albumin and WSO containing 2% egg shell

powder, respectively. Treatment bait was also kept @ 500g/shed

in two rows of ten bait points each. The treatment bait was

exposed to rats for two days after which the remaining bait

was collected and weighed to record the consumption (g/100g

bait) of different types of treatment baits. After one week of

treatment, post census bait consumption was recorded from all

the poultry sheds of each and every block by the same method

as used during pre-census. Reduction in level of rodent

activity (%) with respect to untreated control sheds was

determined as per the formula of Henderson and Tilton (1955)

which is as follows:

Percent reduction in rodent activity = 100 {1−t2 x r1t1 x r2 }

Where, t1 and t2 represent pre- and post-treatment bait

32

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consumptions, respectively in treated sheds and r1 and r2

represent pre- and post-treatment bait consumptions,

respectively in untreated reference sheds.

Reduction in level of rodent activity (%) was also

determined with respect to same shed as per the formula given

below:

Pre-census bait consumption – Post-censusbait consumption x 100

Pre-census bait consumption

3.2.2 Bait additives in traps

Six poultry sheds were selected at the poultry farm

located in the campus of GADVASU, Ludhiana. All the sheds were

predominantly infested with R. rattus. Rodent trapping was

carried out in all the sheds by placing eight single rat traps

in each shed. Each trap was baited with a piece of chapatti.

The effect of 2% poultry egg albumin and 2% egg shell powder

in enhancing rodent trapping was studied by smearing these

components on the chapatti pieces which were then used for

baiting the traps. In first experiment, trapping was carried

out from all the sheds by alternatively placing traps

containing chapatti pieces without additive and those

containing chapatti pieces smeared with 2% egg shell powder.

In the second experiment, trapping was carried out from all

the sheds by alternatively placing traps containing chapatti

pieces without additive and those containing chapatti pieces

smeared with 2% egg albumin. In the third experiment, trapping

was carried out from all the sheds by alternatively placing

traps containing chapatti pieces smeared with 2% egg shell

powder and those containing chapatti pieces smeared with 2%

33

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egg albumin. Every time, trapping was carried out for one week

and the number of rats trapped of each sex was recorded. From

the data on number of rats trapped of each sex, trap index in

the form of total number of rats trapped, number of male rats

trapped and number of female rats trapped per 100 trap nights

per day were calculated.

3.3 Statistical analyses

All values were expressed as mean ± SE. The data on food

consumption for two sexes, three bait additives, three

concentrations of each bait additive, five days of treatment

and five replicated rats was collected using factorial

experiments in Completely Randomized Design. Analysis was done

using General Linear Model in SAS 9.3. All pair wise treatment

comparisons were made using Tukeys’ HSD test at 5% level of

significance.

34

CHAPTER – IV

RESULTS AND DISCUSSION

Present study on evaluation of poultry components (egg

shell and egg albumin) as bait additives for increasing the

acceptance of cereal based bait was carried out in no-choice

and bi-choice feeding tests in laboratory. The effective

concentrations of these components were then mixed in 2% zinc

phosphide bait and tested in poultry farms to evaluate their

acceptance and efficacy in comparison to 2% zinc phosphide

bait without additive. The effect of these poultry components

was also studied on rodent trapping. The results obtained are

presented herewith.

4.1 Laboratory experiments

4.1.1 Bi-choice experiments

In bi-choice laboratory experiments, rats of both sexes

were exposed to WSO bait containing 2, 5 and 10%

concentrations each of ESP, EA and CES. Results revealed no

significant difference in mean daily consumption (g/100g bw)

of WSO bait during pre- and post-treatment periods among all

the treated and untreated groups of rats of both sexes in all

the three experiments using ESP, EA and CES.

4.1.1.1 Acceptance of egg shell powder (ESP) as bait

additive

During treatment period, significantly (P ≤ 0.05) higher

mean daily consumption (g/100g bw) of WSO bait containing 2%

(group I) and 5% (group II) ESP from that of WSO bait alone

was observed in female rats. There was no significant

difference in consumption between WSO bait without additive

and that containing 10% ESP by female rats of treated group

`

III (Table 1, Figure 1). Also there was no significant

difference in mean daily consumption of WSO bait containing

egg shell powder among the three treated groups I, II and III.

Percent acceptance of WSO bait with additive over WSO bait

alone was also not found to differ significantly among the

three treated groups in female rats (Figure 10). The percent

acceptance was found to be 67.21, 58.04 and 49.18%,

respectively in rats of groups I, II and III. Apparently,

there was a decrease in acceptance of bait with increasing

concentration of the bait additive in female rats (Table 1).

In male rat, there was no significant difference observed

in mean daily consumption of WSO bait containing bait additive

and that without any additive at all the concentrations of egg

shell powder tested (Table 2, Figure 2). Also there was no

significant difference in mean daily consumption of WSO bait

containing egg shell powder among the three treated groups I,

II and III. Percent acceptance of WSO bait with additive over

WSO bait alone was also not found to differ significantly

among the three treated groups (Figure 11). The percent

acceptance was found to be 35.19, 41.20 and 47.32%,

respectively in rats of groups I, II and III. Apparently,

there was an increase in acceptance of bait with increasing

concentration of the bait additive in male rats (Table 2).

Percent acceptance of WSO bait containing ESP was

generally low in male rats as compared to female rats. This

difference in acceptance of bait between male and female rats

was found to be statistically significant (P ≤ 0.05) when rats

were exposed to WSO bait containing 2 and 5% ESP in choice

with WSO bait without additive (Figure 3).

36

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Table 1: Acceptance of bait containing egg shell powder by female R. rattus

Group(n =5

each)

Conc.

(%)

Bodywt(g)

Mean daily bait consumption(g/100g bw)

Percentacceptan

cePre-treatme

nt

Duringtreatment

Post-treatme

ntWSObait

WSO +additive

I 2.0 130.00 ±6.32

6.25 ±0.44

2.03±

0.17a

4.02 ±0.21b

5.70 ±0.20

67.21 ±4.88a

II 5.0 130.00 ±9.38

6.52 ±0.62

2.70±

0.16a

4.06 ±0.26b

5.49 ±0.16

58.04 ±5.09a

III 10.0 130.00 ±6.93

7.18 ±0.68

3.87±

0.35a

2.77 ±0.20ab

5.27 ±0.26

49.18 ±5.59a

IV 0.0 130.00 ±8.48

6.61 ±0.37

6.78±

0.52

- 5.81 ±0.24

-

-Values are Mean ± SE, Values with different superscripts (a-b) in a row for during treatment bait consumption and in acolumn for WSO + additive bait consumption and percentacceptance differ significantly at P ≤ 0.05.

Table 2: Acceptance of bait containing egg shell powder by male R. rattus

Group(n =5

each)

Conc.

(%)

Bodywt(g)

Mean daily bait consumption(g/100g bw)

Percentacceptan

cePre-treatme

nt

Duringtreatment

Post-treatme

ntWSObait

WSO +additive

I 2.0 135.00 ±

10.77

4.73 ±0.19

4.36±

0.30a

2.83 ±0.20a

5.07 ±0.10

35.19 ±4.29a

37

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II 5.0 133.00 ± 6.26

5.04 ±0.26

3.44±

0.17a

3.03 ±0.27a

4.94 ±0.03

41.20 ±4.39a

III 10.0 135.00 ±

10.20

5.37 ±0.43

5.01±

0.64a

3.93 ±0.27a

6.41 ±0.25

47.32 ±6.34a

IV 0.0 141.00 ± 6.39

5.76 ±0.40

5.85±

0.27

- 4.13 ±0.08

-

-Values are Mean ± SE, Values with similar superscript (a) ina row for during treatment bait consumption and in a columnfor WSO + additive bait consumption and percent acceptance donot differ significantly.

2% 5% 10%0

0.51

1.52

2.53

3.54

4.5 FemaleWSO WSO + Additive

Conc. of egg shell powder

Mean

dai

ly b

ait

cons

umpt

ion

(g/1

00g

bw)

a

b

b

a

a

a

Figure 1. Comparison of mean daily bait consumption by femalerats between WSO bait and WSO bait containingdifferent concentrations of egg shell powder in bi-choice feeding tests.

38

`

2% 5% 10%0

1

2

3

4

5

6Male WSO WSO + Additive


Mean

dai

ly b

ait

cons

umpt

ion

(g/1

00g

bw)

a

aa

a

a

a

Figure 2. Comparison of mean daily bait consumption by malerats between WSO bait and WSO bait containingdifferent concentrations of egg shell powder in bi-choice feeding tests.

2% 5% 10%0

10

20

30

40

50

60

70

80 FemaleMale


Perc

ent

acce

ptan

ce

a

b

a

b a a

39

`

Figure 3. Comparison of percent acceptance of baitcontaining different concentrations of egg shellpowder between male and female rats in bi-choicefeeding tests

Table 3: Acceptance of bait containing egg albumin by female R.

rattus

Group(n =5

each)

Conc.

(%)

Bodywt(g)

Mean daily food consumption(g/100g bw)

Percentacceptan

cePre-

treatment

Duringtreatment

Post-treatme

ntWSObait

WSO +additi

ve

I 2.0 131.00± 0.89

11.05 ±0.43

6.39 ±0.79a

10.04± 0.32b

8.92 ±0.46

67.45 ±3.83a

II 5.0 136.00±

14.31

12.77 ±1.03

7.25 ±0.45a

13.04± 0.57b

11.21 ±0.21

69.13 ±4.55a

III 10.0 134.00±

14.58

12.95 ±0.24

8.09 ±0.58a

12.71± 0.52b

8.72 ±0.38

65.00 ±3.86a

IV 0.0 127.00± 6.87

11.15 ±0.17

10.52± 0.52

- 9.39 ±0.37

-


40

`

Table 4: Acceptance of bait containing egg albumin by male R.

rattus

Group(n =5

each)

Conc.

(%)

Bodywt(g)


Percentacceptan

cePre-

treatment

Duringtreatment

Post-treatme

ntWSObait

WSO +additi

ve

I 2.0 130.00± 5.65

12.70 ±0.81

8.17 ±0.60a

9.98 ±0.40ab

10.61 ±0.75

57.12 ±4.36a

II 5.0 138.00±

17.06

9.58 ±0.40

5.79 ±0.32a

10.97± 0.05b

9.01 ±0.14

70.63 ±3.81b

III 10.0 132.00± 3.34

10.49 ±0.49

4.69 ±0.42a

11.57± 0.48b

7.88 ±0.11

76.22 ±4.11b

IV 0.0 123.00± 3.89

10.33 ±0.46

9.94 ±0.41

- 8.79 ±0.34

-


41

`

2% 5% 10%0

2

4

6

8

10

12

14 FemaleWSO WSO + Additive

Conc. of egg albumin

Mean

dai

ly b

ait

cons

umpt

ion

(g/1

00g

bw)

a aa

bb

b

Figure 4. Comparison of mean daily bait consumption by femalerats between WSO bait and WSO bait containingdifferent concentrations of egg albumin in bi-choicefeeding tests.

2% 5% 10%0

2

4

6

8

10

12

14 Male WSO WSO + Additive


Mean

dai

ly b

ait

cons

umpt

ion

(g/1

00g

bw)

a

a

aa

bb

42

`

Figure 5. Comparison of mean daily bait consumption by malerats between WSO bait and WSO bait containingdifferent concentrations of egg albumin in bi-choicefeeding tests

2% 5% 10%0

102030405060708090

Female Male


Perc

ent

acce

ptan

ce

a

a

a aa

a

Figure 6. Comparison of percent acceptance of bait containingdifferent concentrations of egg albumin between maleand female rats in bi-choice feeding tests

4.1.1.3. Acceptance of crushed egg shell (CES) as bait

additive

No significant difference in mean daily consumption

between WSO bait containing 2, 5 and 10% CES and WSO bait

without additive was observed in female rats. The consumption

of two kinds of baits was almost similar (Table 5, Figure 7).

Also there was no significant difference in mean daily

consumption of WSO bait containing CES among the three treated

groups I, II and III. Percent acceptance of WSO bait with

additive was also not found to differ significantly among the

three treated groups (Figure 10). The percent acceptance was

43

`

found to be 55.34, 54.25 and 53.62%, respectively in rats of

groups I, II and III (Table 5).

In male rats also no significant difference in mean daily

consumption between WSO bait containing 2, 5 and 10% CES and

WSO bait without additive was observed. The consumption of two

kinds of baits was almost similar (Table 6, Figure 8). Also

there was no significant difference in mean daily consumption

of WSO bait containing CES among the three treated groups I,

II and III. Percent acceptance of WSO bait with additive was

also not found to differ significantly among the three treated

groups (Figure 11). The percent acceptance was found to be

62.39, 55.35 and 58.94%, respectively in rats of groups I, II

and III (Table 6).

Percent acceptance of WSO bait containing CES by male

rats as compared to female rats was generally high at all the

three concentrations of CES tested. This difference in

acceptance of bait between male and female rats was, however,

not found to differ significantly at any of the concentrations

tested (Figure 9).

Table 5: Acceptance of bait containing crushed egg shells by female R. rattus

Group(n =5

each)

Conc.

(%)

Bodywt(g)


Percentacceptan

cePre-

treatment

Duringtreatment

Post-treatme

ntWSObait

WSO +additi

ve

I 2.0 143.00± 2.68

12.85 ±0.37

7.00±

0.42a

8.67 ±0.47a

11.54 ±0.29

55.34 ±6.10a

II 5.0 144.00 14.66 ± 8.28 9.67 ± 11.78 ± 54.25 ±

44

`

± 2.19 0.51 ±0.65a

0.62a 0.18 4.16a

III 10.0 142.00± 1.79

12.88 ±0.42

6.61±

0.49a

7.13 ±0.52a

10.99 ±0.36

53.62 ±3.62a

IV 0.0 146.00±

10.80

14.20 ±0.74

15.02±

0.40

- 11.09 ±0.18

-

-Values are Mean ± SE, Values with similar superscript (a) ina row for during treatment bait consumption and in a columnfor WSO + additive bait consumption and percent acceptance donot differ significantly.

Table 6: Acceptance of bait containing crushed egg shells by male R. rattus

Group(n =5

each)

Conc.

(%)

Bodywt(g)


Percentacceptan

cePre-

treatment

Duringtreatment

Post-treatme

ntWSObait

WSO +additi

ve

I 2.0 149.00± 5.72

11.17 ±0.63

7.62±

0.78a

9.92 ±0.30a

10.28 ±0.24

62.39 ±4.69a

II 5.0 149.00± 5.72

13.89 ±0.33

8.18±

0.64a

8.98 ±0.48a

13.84 ±0.42

55.35 ±5.07a

III 10.0 146.00± 7.79

13.12 ±0.37

7.08±

0.39a

10.46±

0.75a

11.82 ±0.39

58.94 ±3.31a

IV 0.0 144.00± 4.56

13.85 ±0.11

12.92±

0.26

- 12.36 ±0.42

-

-Values are Mean ± SE, Values with similar superscript (a) ina row for during treatment bait consumption and in a column

45

`

for WSO + additive bait consumption and percent acceptance donot differ significantly.

2% 5% 10%0

2

4

6

8

10

12 FemaleWSO WSO + Additive

Conc. of crushed egg shells

Mean

dai

ly b

ait

cons

umpt

ion

(g/1

00g

bw)

a

a aa

aa

Figure 7. Comparison of mean daily bait consumption by femalerats between WSO bait and WSO containing differentconcentrations of crushed egg shells in bi-choicefeeding tests.

46

`

2% 5% 10%0

2

4

6

8

10

12Male WSO WSO + Additive


Mean

dai

ly b

ait

cons

umpt

ion

(g/1

00g

bw)

a

a

aa

a

a

Figure 8. Comparison of mean daily bait consumption by malerats between WSO bait and WSO bait containingdifferent concentrations of crushed egg shells in bi-choice feeding tests.

2% 5% 10%48

50

52

54

56

58

60

62

64FemaleMale


Perc

ent

acce

ptan

ce

aa

a a aa

47

`

Figure 9. Comparison of percent acceptance of bait containingdifferent concentrations of crushed egg shells betweenmale and female rats in bi-choice feeding tests.

ESP EA CES0

10

20

30

40

50

60

70

80 Female2% 5% 10%

Perc

ent

acce

ptan

ce

aa

a

a aa

a a a

Figure 10. Comparison of percent acceptance of bait amongdifferent concentrations of egg shell powder, eggalbumin and crushed egg shells in female rats in bi-choice feeding tests.

48

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ESP EA CES0

10

20

30

40

50

60

70

80

90 Male2% 5% 10%

Perc

ent

acce

ptan

ce

aa

aa

bb

aa a

Figure 11. Comparison of percent acceptance of bait amongdifferent concentrations of egg shell powder, eggalbumin and crushed egg shells in male rats in bi-choice feeding tests.

In female rats, the percent acceptance of WSO bait

containing 2% ESP, EA and CES was not found to differ

significantly. However, the percent acceptance of WSO bait

containing 5% EA was significantly (P ≤ 0.05) high from WSO

bait containing 5% CES and non-significantly high from WSO

bait containing 5% ESP (Figure 12). The percent acceptance of

WSO bait containing 10% EA was significantly (P ≤ 0.05) high

from WSO bait containing 10% ESP and non-significantly high

from WSO bait containing 10% CES (Figure 12). There was no

significant difference in percent acceptance of WSO bait

containing ESP and CES at all the concentrations tested.

In male rats, percent acceptance of WSO bait containing

EA and CES was found to be significantly (P ≤ 0.05) high from

that of WSO bait containing ESP at all the concentrations

49

`

tested (Figure 13). At 5 and 10% concentrations, percent

acceptance of WSO bait containing EA was comparatively more

than WSO bait containing similar concentrations of CES.

Statistically, there was no significant difference in percent

acceptance of WSO bait containing EA and CES.

In overall, the percent acceptance of WSO bait containing

2 and 5% ESP and 2, 5 and 10% EA was significantly more than

WSO bait alone in female rats. Whereas, in male rats, the

percent acceptance of WSO bait containing 2 and 5% EA was

significantly more than WSO bait alone. So from present

studies it can be concluded that addition of EA in WSO bait

increases the acceptance of bait by both male and female rats.

This may be due to high nutritional value of EA than ESP and

CES.

2% 5% 10%0

10

20

30

40

50

60

70

80Female WSO + ESP

WSO + EA

Conc.of bait additives

Perc

ent

acce

ptan

ce

a a

a ab

a

ba

b

ab

Figure 12. Comparison of percent acceptance of baitscontaining different bait additives by female rats inbi-choice feeding tests.

50

`

2% 5% 10%0

102030405060708090

Male WSO + ESPWSO + EA

Conc. of bait additives

Perc

ent

acce

ptan

ce

a

bb

a

b

b

b

ba

Figure 13. Comparison of percent acceptance of baitscontaining different bait additives by male rats inbi-choice feeding tests.

Taste of food plays a significant role in food preference

by R. rattus (Kandhwal 2009). The preference of rats for a

particular food depends upon its flavour more than its

nutritional value (Naganuma et al 1973). As suggested by Young(1946), the criteria for food selection in rats are complex

and may depend upon many factors. Likewise, Jackson (1965)

also revealed that texture, odour and taste play a significant

role in selection of a particular bait base. According to

Marsh (1986), taste often supersedes any earlier influence of

odour in food selection to a degree that is not paralleled in

humans. Behavioral characteristics of rodents have revealed

that their choice for food depends upon many factors including

caloric value (Hausmann 1932), deliciousness (Young 1946) and

behavior modifying components (Barnett 1956). Some food items

51

`

are intermittently preferred because of the energy they

provide (Stenseth 1977), chiefly due to their carbohydrate and

protein contents (Smythe 1976). These findings support present

results where we observed huge variations among the food

preferences for different taste enhancers. Like, there was no

significant difference observed between the consumption of WSO

bait alone and that containing 2, 5 and 10% crushed egg shell

by both male and female rats. There was no significant

difference observed between consumption of WSO bait alone and

that containing 2, 5 and 10% egg shell powder by male rats,

however, a significantly higher consumption of WSO bait

containing 2 and 5% egg shell powder was observed by female

rats. Also significantly higher consumption was observed of

WSO bait containing 5 and 10% egg albumin by male rats and

that of WSO containing all the three concentrations of egg

albumin by female rats. Present results thus indicate sex

specific differences in acceptance of cereal based bait

containing different poultry egg components as bait additives

in bi-choice. Similar sex specific variations in response of R.

rattus have also been reported earlier (Kaur et al 2008).

4.1.2 No-choice experiments under FSCM

In female rats (110-155g bw), no significant difference

was observed in average consumption (g/kg bw) of bait in a

period of 2 hr among WSO bait without additive and WSO baits

containing 2% concentration each of ESP, EA and CES in no-

choice feeding tests under FSCM (Table 7, Figure 14). However,

the average consumption of WSO bait with all the three types

of additives was non-significantly high than WSO bait without

additive. Further, among the three treated baits, the average

consumption of bait containing 2% EA was non-significantly

52

`

high than the other two bait additives. The total number of

bouts made by female rats in 2 hr towards WSO bait without

additive and those with three bait additives were also not

found to differ significantly (Table 7, Figure 14). The

number of bouts by female rats were highest towards bait

containing 2% EA and least towards bait containing 2% CES.

Total time spent (s) in bouting in 2 hr a day by female rats

was also maximum in bait containing 2% EA and least in bait

containing 2% CES (Table 7). The total time spent by rats in 2

hr among WSO bait without additive and baits containing three

additives was also not found to differ significantly (Figure

14). Also there was no significant difference in total time

spent towards bouting among the four replicated rats when

exposed to both treated and untreated baits.

Table 7. Acceptance of bait containing different additives byfemale R. rattus as recorded under FSCM

Parameters WSO bait WSO +2% ESP WSO + 2% EA WSO + 2% CES

Foodconsumptio

n(g/kg bw)

1.96±0.49a 4.01±1.00a 6.33±1.58a 6.01±1.50a

Number ofbouts made

4.85±1.48a 4.50±1.43a 6.20± 3.16a 2.45±0.51a

Time spenttowardsbouting(s)

175.10±78.38a

156.50±63.39a 181.50±92.21a

69.10±18.88a

-Values are Mean ± SE, Values with similar superscript (a) ina row do not differ significantly.

53

`

Food consumption (g/kg...

Number of bouts Time spent (s)0

20406080

100120140160180200

FemaleWSO WSO + 2% ESP

a a a a a

a

aa

a

a a a

Figure 14. Comparison of response of female rats towardsdifferent baits in no-choice feeding tests.

In male rats ( 155-205g bw) also, no significant

difference was observed in average consumption (g/kg bw) of

bait in 2 hr period among WSO bait without additive and WSO

baits containing 2% concentration each of ESP, EA and CES in

no-choice feeding tests under FSCM (Table 8, Figure 15). The

average consumption of WSO bait with additive was non-

significantly less than WSO without additive. Among the three

baits with additives, the average consumption of WSO bait

containing 2% EA and CES was non-significantly high than bait

containing 2% ESP. The total number of bouts made by male rats

in 2 hr period towards WSO bait without additive were

significantly (P ≤ 0.05) more than those made towards baits

containing three types of additives (Table 8, Figure 15). No

significant difference was, however, found in the number of

54

`

bouts among the WSO baits containing 2% ESP, EA and CES. Total

time spent (s) in bouting in 2 hr period a day by male rats

was also significantly (P ≤ 0.05) more in WSO without any

additive than baits containing three additives (Table 8,

Figure 15). No significant difference was, however, found in

the time spent among the WSO baits containing 2% ESP, EA and

CES. The time spent for bouting in the baits containing 2% ESP

and EA was non-significantly more than that spent in bait

containing 2% CES.

Table 8:Acceptance of bait containing different additives bymale R. rattus as recorded under FSCM

Parameters WSO bait WSO +2% ESP WSO + 2% EA WSO + 2% CES

Foodconsumptio

n(g/kg bw)

7.28±1.82a 2.61±0.65a 6.04±1.51a 6.22±1.55a

Number ofbouts made

12.55±4.35a 4.10 ±1.22b 4.40±0.56b 2.80±0.19b

Time spenttowardsbouting(s)

689.45±350.08a

172.45±77.12b

163.35±42.76b

99.05±14.30b

-Values are Mean ± SE, Values with different superscripts (a-b) in a row differ significantly at P ≤ 0.05.

55

`

Food consumption (g/kg bw)

Number of bouts Time spent (s)0

100

200300

400

500

600

700800

MaleWSO WSO + 2% ESPWSO + 2% EA WSO + 2% CES

a a a a a b b bb

bb

a

Figure 15. Comparison of response of male rats towardsdifferent baits in no-choice feeding tests.

In overall, no significant difference was found in

average food consumption (g/kg bw) for 2 hr a day between male

and female rats (Figure 16) in respect of WSO bait without

additive as well as of WSO bait containing 2% ESP, EA and CES.

The number of bouts made towards WSO bait without additive

were significantly (P ≤ 0.05) more by male rats compared to

female rats (Figure 17). However, there was no significant

difference in number of bouts made towards WSO bait containing

three bait additives between male and female rats. The time

spent in bouting towards WSO bait without additive was also

significantly (P ≤ 0.05) more by male rats compared to female

rats (Figure 18) and there was no significant difference in

56

`

time spent in bouting towards WSO bait containing three bait

additives between male and female rats.

These results under FSCM thus indicate almost similar

consumptions of WSO bait without additive and those with

additives in no-choice tests. Among baits with different

additives, however, the bait consumption, number of bouts made

and the time spent were non-significantly more towards the

bait containing 2% EA than baits containing 2% ESP and CES

which may again be due to high nutritional value of EA as

compared to ESP and CES. The results of no-choice feeding

tests under FSCM also indicate sex specific differences in

behaviour of R. rattus regarding consumption of WSO bait

containing poultry egg components as bait additives, number of

bouts made as well as the time spent.

WSO WSO + 2% ESP WSO + 2% EA WSO + 2% CES0

1

2

3

4

5

6

7

8FemaleMale

Food

con

sume

d (g

/kg

bw)

a

a

a

a

a a a a

57

`

Figure 16. Comparison of total consumption of different baitsin 2 hr period between male and female rats in no-choice feeding tests.


2

4

6

8

10

12

14 FemaleMale

Numb

er o

f bo

uts

a

b

a a a

a

a a

Figure 17. Comparison of total number of bouts made towardsdifferent baits in 2 hr period between male and femalerats in no-choice feeding tests.

58

`


100

200

300

400

500

600

700

800 FemaleMale

Time

spe

nt (

s)

a

b

a a aa

a a

Figure 18. Comparison of total time spent towards bouting indifferent baits in 2 hr period between male and femalerats in no-choice feeding tests.

4.2 Experiments in poultry farms

4.2.1 Bait additives in rodenticide baits

Three blocks (I, II and III) of poultry farm selected at

village Ghutani Kalan, District Ludhiana were treated with 2%

zinc phosphide bait, 2% zinc phosphide bait containing 2% egg

albumin and 2% zinc phosphide bait containing 2% egg shell

powder, respectively. Block IV selected at the Campus,

GADVASU, Ludhiana was kept as untreated control. All the sheds

were predominantly infested with R. rattus. Rats were found

sitting on the roof of the poultry sheds, making burrows in

the dried excreta lying below the shed and damaging poultry

feed as well as eggs (Plates 2 and 3). Pre-census bait

consumption (g/100g bait) recorded before treatment from all

four blocks by keeping plain WSO bait revealed 29.30 to 55.80%

59

`

consumption (Table 9). No significant difference was found in

pre-census bait consumption among the four treated and

untreated blocks indicating almost similar level of rodent

activity in all the blocks (Figure 19). The consumption of

treatment bait was found to be 26.95, 68.20 and 68.65% in

blocks I, II and III, respectively (Table 9). The consumption

of 2% zinc phosphide bait containing 2% egg albumin and 2% egg

shell powder was found to be significantly (P ≤ 0.05) higher

than that of 2% zinc phosphide bait alone (Figure 19).

Comparatively less consumption of zinc phosphide bait without

additive in block I may be due to neophobia behaviour shown by

rats leading to bait aversion towards highly toxic rodenticide

bait. Record of post census bait consumption after one week of

treatment from all the blocks revealed a decrease in bait

consumption from that observed during pre census in all the

three treated blocks, whereas, an increase in bait consumption

from that pre-census bait consumption was observed in

untreated block.

Table 9. Census and treatment bait consumption in differentblocks of poultry farms

Block Treatment Bait consumption (g/100g bait)

Pre-census Treatment Post-census

I ZnP 29.30±11.89a 26.95±13.99a 23.75±10.82

II ZnP + EA 55.60±7.74a 68.20±8.86b 42.20±3.19

III ZnP + ESP 55.80±13.42a 68.65±13.95b 45.70±8.56

IV Control 33.40±8.70a - 38.15±4.65

Values are Mean±SE, ZnP = 2% Zinc phosphide, EA = 2% Egg albumin, ESP = 2% Egg shell powder, Values with different

60

`

superscripts (a-b) in a column differ significantly at P ≤ 0.05.

Pre-census Treatment0

10

20

30

40

50

60

70

80ZnP ZnP + EA ZnP + ESP UT

Bait

con

sump

tion

(g/

100g

ba

it)

a

aa

a a

bb

Figure 19. Comparison of bait consumptions during census andtreatment in different poultry blocks.

The reduction in rodent activity in the three treated

blocks was found to vary from 28.13 to 44.41% with respect to

untreated block and 19.23 to 42.69% with respect to same shed

(Table 10). There was no significant difference in reduction in

rodent activity between the three treated blocks (Figure 20),

though apparently, there was a higher decrease in rodent

activity in block III treated with 2% zinc phosphide bait

containing 2% ESP as additive. Results in poultry farm therefore

reveal higher efficacy of 2% zinc phosphide containing 2% ESP.

Less efficacy of rodenticide treatment in poultry farm during

present studies may be due to the availability of other food

items such as poultry feed, eggs, chicks etc in the poultry farm

which may further have led to reduced acceptance of rodenticide

bait. Present results, however, suggest the potential of EA and

ESP in increasing the acceptance of 2% zinc phosphide bait.

61

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Table 10. Reduction in rodent activity in treated blocks of

poultry farm

Block Treatment

Reduction in rodent activity (%)

With respect tountreated block

With respect to sameblock

I ZnP 33.52±3.46a 24.07±3.95a

II ZnP + EA 28.13±8.64a 19.23±9.71a

III ZnP +ESP

44.41±6.81a 42.69±9.58a

IV untreated

- Increase in activity

Values are Mean±SE, ZnP = 2% Zinc phosphide, EA = 2% Eggalbumin, ESP = 2% Egg shell powder, Values with similar superscript (a) in a column do not differ significantly.

With respect to untreated b...

With respect to same block05

101520253035404550 ZnP ZnP + EA ZnP + ESP

Redu

ctio

n in

rod

ent

acti

vity

(%

)

a a

a

a a

a

Figure 20. Comparison of reduction in rodent activity aftertreatment with rodenticide bait containing baitadditives.

62

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The literature also reveals the use of cereals in whole-

some or cracked form mixed with additives such as vegetable

oil, egg shell, egg yolk, minced meat, sugar and flavours

(Shafi et al 1990, Pervez et al 2003). Khan et al (2000) reported

preference for poison bait containing egg shells by rodents in

rice crop fields. Pervez et al (2005) found highest potential of

egg mixed brodifacoum bait in enhancing acceptance by field

rodents in wheat crop. However, the addition of egg shell at

2% did not show a significant increase in relative consumption

for the cereal base by B. bengalensis (Naeem et al 2011). Rather

they observed an inverse trend indicating decreased preference

for plain millet based bait mixed with 5% egg shell. Shafi et al

(1993) also observed that B. bengalensis show a high preference

toward minced meat bait than egg shell and egg yolk mixed

bait. Mushtaq et al (2013) also did not found any increase in

acceptance of cereal base containing egg shell by Hystrix indica.

However, Pervez et al (1999 and 2003) found an additive effect of

2% egg shell in enhancement of bait preference. These

contradictory results may primarily be attributed to the

nature of bait material used and due to the different species

tested.

4.2.2 Bait additives in trapsRodent trapping with wooden single rat traps was carried

out in sheds of poultry farm located in the campus of GADVASU,Ludhiana. Each trap was baited with a piece of chapattismeared with 2% (by weight) poultry ESP and 2% EA. For thefirst time, trapping was carried out from all the sheds byalternatively placing traps containing chapatti pieces withoutadditive and those containing chapatti pieces smeared with 2%

63

`

egg shell powder. Results revealed total trap index of R. rattus

to be 11.60 (6.84 for male and 4.76 for female rats) in trapsbaited with chapatti piece smeared with ESP and 13.38 (6.84for male and 6.54 for female rats) in traps baited withchapatti piece without any additive (Table 11). No significantdifference was found in total trap index as well as that ofmale and female rats separately between the traps baited withchapatti pieces with and without ESP (Figure 21). Also nosignificant difference was found in the trap index of trapsbaited with chapatti pieces smeared with and without ESPbetween the two sexes (Figure 22). Table 11. Comparison of trap index of rats trapped by smearing

chapatti pieces with and without egg shell powder asadditive

Additive used onchapatti

Trap index(no. of animals trapped/100 traps/day)

Male Female Total

Egg shell powder 6.84 ±2.04 a 4.76 ±1.87 a 11.60± 3.69 a

Untreated 6.84 ±1.22 a 6.54 ±2.87 a 13.38± 3.61 a

Values are Mean ±SE, Values with similar superscript (a) in acolumn and in a row for two sexes do not differ significantly.

64

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Male Female Total0

2

4

6

8

10

12

14

16Egg shell powder

Trap

ind

ex

a aa

a

aa

Figure 21. Comparison of trap index of rats trapped bysmearing chapatti pieces with and without egg shellpowder as additive.

Egg shell powder Untreated0

1

2

3

4

5

6

7

8 MaleFemale

Trap

ind

ex

a

a

a

a

Figure 22. Comparison of trap index of male and female ratstrapped by smearing chapatti pieces with and withoutegg shell powder as additive.

65

`

For the second time, trapping was carried out from all

the sheds by alternatively placing traps containing chapatti

pieces without additive and those containing chapatti pieces

smeared with 2% EA. Results revealed total trap index of R.

rattus to be 5.35 (3.27 for male and 2.08 for female rats) in

traps baited with chapatti piece smeared with EA and 7.13

(2.08 for male and 5.05 for female rats) in traps baited with

chapatti piece without any additive (Table 12). No significant

difference was found in total trap index as well as that of

male and female rats separately between the traps baited with

chapatti pieces with and without EA (Figure 23). Also no

significant difference was found in the trap index of traps

baited with chapatti pieces smeared with and without EA

between the two sexes (Figure 24).

Table 12. Comparison of trap index of rats trapped by smearingchapatti pieces with and without egg albumin asadditive



Male Female Total

Egg albumin 3.27 ±1.06 a 2.08 ±0.78 a 5.35 ±1.68 a

Untreated 2.08 ±1.29 a 5.05 ±1.42 a 7.13 ±2.59 a


66

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Male Female Total0

1

2

3

4

5

6

7

8 Egg albuminUntreated

Trap

ind

ex

aa

a

aa

a

Figure 23. Comparison of trap index of rats trapped bysmearing chapatti pieces with and without egg albuminas additive.

Egg albumin Untreated0

1

2

3

4

5

6 Male

Trap

ind

ex

a

aa

a

67

`

Figure 24. Comparison of trap index of male and female ratstrapped by smearing chapatti pieces with and withoutegg albumin as additive.

For the third time, trapping was carried out from all the

sheds by alternatively placing traps containing chapatti

pieces smeared with 2% ESP and 2% EA. Results revealed total

trap index of R. rattus to be 13.69 (6.25 for male and 7.44 for

female rats) in traps baited with chapatti piece smeared with

ESP and 12.79 (8.03 for male and 4.76 for female rats) in

traps baited with chapatti piece smeared with EA (Table 13).

No significant difference was found in total trap index as

well as that of male and female rats separately between the

traps baited with chapatti pieces smeared with ESP and EA

(Figure 25). Also no significant difference was found in the

trap index of traps baited with chapatti pieces smeared with

ESP and EA between the two sexes (Figure 26).

Table 13. Comparison of trap index of rats trapped by smearingchapatti pieces with egg shell powder and egg albuminas additives



Male Female Total

Egg shell powder 6.25 ±2.01 a 7.44 ±2.61 a 13.69± 4.12 a

Egg albumin 8.03 ±2.22 a 4.76 ±1.55 a 12.79± 2.03 a


68

`

Male Female Total0

2

4

6

8

10

12

14

16 Egg shell ...

Trap

ind

ex

aa a

a

a

a

Figure 25. Comparison of trap index of rats trapped bysmearing chapatti pieces with egg shell powder and eggalbumin as additives.

Egg shell powder Egg albumin0

1

2

3

4

5

6

7

8

9 Male Female

Trap

ind

ex

a

a a

a

Figure 26. Comparison of trap index of male and female ratstrapped by smearing chapatti pieces with egg shellpowder and egg albumin as additives.

69

`

In overall, the results of poultry trapping experiments

revealed no significant effect of poultry egg components i.e.

ESP and EA in trapping of rats by single catch rat traps. This

may be due to the reason that rats might not have come to know

about the additive smeared on chapatti piece before they

pulled it from the string of the trap and got trapped. Due to

this very reason there was no significant difference in trap

index between traps baited with chapatti piece smeared with

and without additive and also between the traps baited with

chapatti smeared with ESP and EA.

During present studies, results of laboratory experiments

in bi-choice revealed significantly (P ≤ 0.05) higher

consumption of WSO bait containing all the three

concentrations (2, 5 and 10%) of EA and 2 and 5% ESP by female

rats and that of bait containing 5 and 10% EA by male rats. In

no-choice experiments, the bait consumption, number of bouts

made and the time spent in WSO bait containing 2 % EA by

female rats was non-significantly high than that containing 2%

ESP and CES. In male rats, the results were, however,

inconclusive. In poultry farm also significantly higher

consumption of 2% zinc phosphide bait containing 2% EA and 2%

ESP leading to upto 44.41% reduction in rodent activity was

observed compared to 33.52% reduction in rodent activity with

only 2% zinc phosphide treatment.

We may therefore assume that use of 2% egg albumin and

egg shell powder in cereal based bait may enhance bait intake

as compared to plain bait. This combination may yield

significant control of R. rattus if used in 2% zinc phosphide

bait.

70

CHAPTER – V

SUMMARY

Present study on evaluation of poultry components i.e.

egg shell powder (ESP), egg albumin (EA) and crushed egg

shells (CES) as bait additives for increasing the acceptance

of cereal based bait by house rat, Rattus rattus was carried out

in bi-choice and no-choice laboratory feeding tests. The

effective concentrations of these components were then tested

in poultry farms to evaluate the acceptance and efficacy of 2%

zinc phosphide bait without additive and that containing 2%

ESP and EA. The effect of these poultry components was also

studied on rodent trapping.

In bi-choice laboratory experiments, no significant

difference was observed in mean daily consumption (g/100g bw)

of WSO bait during pre- and post-treatment periods among all

the treated and untreated groups of rats of both sexes for all

the bait additives tested. During treatment with different

concentrations of ESP, significantly (P ≤ 0.05) higher mean

daily consumption (g/100g bw) of WSO bait containing 2 and 5%

ESP from that of WSO bait alone was observed in female rats.

There was no significant difference in consumption between WSO

bait without additive and that containing 10% ESP by female

rats. Also there was no significant difference in mean daily

consumption of WSO bait containing egg shell powder among the

three treated groups. Percent acceptance of WSO bait with

additive over WSO bait alone was also not found to differ

significantly in female rats among the three treated groups.

The percent acceptance was found to be 67.21, 58.04 and

49.18%, respectively in rats of groups treated with 2, 5 and

10% ESP.

`

In male rat, there was no significant difference observed

in mean daily consumption of WSO bait containing bait additive

and that without any additive at all the concentrations of ESP

tested. Also there was no significant difference in mean daily

consumption of WSO bait containing ESP among the three treated

groups. Percent acceptance of WSO bait with additive over WSO

bait alone was also not found to differ significantly among

the three treated groups. The percent acceptance was found to

be 35.19, 41.20 and 47.32%, respectively in rats of groups

treated with 2, 5 and 10% ESP.

Percent acceptance of WSO bait containing ESP was

generally low in male rats as compared to female rats. This

difference in acceptance of bait between male and female rats

was found to be statistically significant (P ≤ 0.05) when rats

were exposed to WSO bait containing 2% ESP in choice with WSO

bait without additive.

During treatment with different concentrations of EA,

significantly (P ≤ 0.05) higher mean daily consumption of WSO

bait containing 2, 5 and 10% EA from that of WSO without

additive was observed in female rats. There was no significant

difference in mean daily consumption of WSO bait containing EA

among the three treated groups. Percent acceptance of WSO bait

with additive over WSO bait alone was also not found to differ

significantly in female rats among the three treated groups.

The percent acceptance was found to be 67.45, 69.13 and 65%,

respectively in rats of groups treated with 2, 5 and 10% EA.

In male rat, significantly (P ≤ 0.05) higher mean daily

consumption of WSO bait containing 5 and 10% EA was found from

that of WSO bait alone. There was no significant difference

between the consumption of WSO bait without additive and that

72

`

containing 2% EA. Also there was no significant difference in

mean daily consumption of WSO bait containing EA among the

three treated groups. Percent acceptance of WSO bait with

additive over WSO bait alone was found to be significantly (P

≤ 0.05) high in rats of groups treated with 5% (70.63%) and

10% (76.22%) EA as compared to rats of group treated with 2%

EA (57.12%).

Percent acceptance of WSO bait containing EA by male rats

as compared to female rats was generally low at 2% EA and high

at 5 and 10% EA. This difference in acceptance of bait between

male and female rats was, however, found to be statistically

non-significant at all the concentrations tested.

During treatment with different concentrations of CES, no

significant difference in mean daily consumption between WSO

bait containing 2, 5 and 10% CES and WSO bait without additive

was observed in both male and female rats. The consumption of

two kinds of baits was almost similar. Also there was no

significant difference in mean daily consumption of WSO bait

containing CES among the three treated groups. Percent

acceptance of WSO bait with additive was also not found to

differ significantly among the three treated groups. The

percent acceptance was found to be 55.34, 54.25 and 53.62%,

respectively in female rats and 62.39, 55.35 and 58.94%,

respectively in male rats of groups treated with 2, 5 and 10%

CES.

Percent acceptance of WSO bait containing CES by male

rats as compared to female rats was generally high at all the

three concentrations of CES tested. This difference in

acceptance of bait between male and female rats was, however,

73

`

not found to differ significantly at any of the concentrations

tested.

In female rats, the percent acceptance of WSO bait

containing 2% ESP, EA and CES was not found to differ

significantly. However, the percent acceptance of WSO bait


bait containing 5% CES and non-significantly high from WSO

bait containing 5% ESP. The percent acceptance of WSO bait


bait containing 10% ESP and non-significantly high from WSO

bait containing 10% CES. There was no significant difference

in percent acceptance of WSO bait containing ESP and CES at

all the concentrations tested.

In male rats, percent acceptance of WSO bait containing

EA and CES was found to be significantly (P ≤ 0.05) high from

that of WSO bait containing ESP at all the concentrations

tested. At 5 and 10% concentrations, percent acceptance of WSO

bait containing EA was comparatively more than WSO bait

containing similar concentrations of CES. Statistically, there

was no significant difference in percent acceptance of WSO

bait containing EA and CES. Present results thus indicate sex

specific differences in acceptance of cereal based bait

containing different poultry egg components as bait additives

in bi-choice. So from present studies it can be concluded that

addition of EA in WSO bait increases the acceptance of bait by

both male and female rats. This may be due to high nutritional

value of EA than ESP and CES.

In no-choice experiments under food scale consumption

monitor, no significant difference was observed in average

consumption (g/kg bw) of bait for 2 hr period by female rats

74

`

among WSO bait without additive and WSO baits containing 2%

concentration each of ESP, EA and CES. The average consumption

of WSO bait with all the three types of additives was non-

significantly high than WSO bait without additive. Among the

three treated baits, the average consumption of bait

containing 2% EA was non-significantly high than other two

bait additives. The total number of bouts made by female rats

in 2 hr period towards WSO bait without additive and those

with three bait additives were also not found to differ

significantly. The number of bouts by female rats were highest

towards bait containing 2% EA and least towards bait

containing 2% CES. Total time spent (s) in bouting in 2 hr

period by female rats was also maximum in bait containing 2%

EA and least in bait containing 2% CES. The total time spent

by rats in 2 hr among WSO bait without additive and baits

containing three additives was also not found to differ

significantly.

In male rats also, no significant difference was observed

in average consumption (g/kg bw) of bait for 2 hr period among

WSO bait without additive and WSO baits containing 2%

concentration each of ESP, EA and CES in no-choice feeding

tests under FSCM. The average consumption of WSO bait with

additive was non-significantly less than WSO without additive.

Among the three baits with additives, the average consumption

of WSO bait containing 2% ESP and EA was non-significantly

high than bait containing 2% ESP. The total number of bouts

made by male rats in 2 hr period towards WSO bait without

additive were significantly (P ≤ 0.05) more than those made

towards baits containing three types of additives. No

significant difference was, however, found in the number of

75

`

bouts among the WSO baits containing 2% ESP, EA and CES. Total

time spent (s) in bouting in 2 hr period a day by male rats

was also significantly (P ≤ 0.05) more in WSO without any

additive than baits containing three additives. No significant

difference was found in the time spent among the WSO baits

containing 2% ESP, EA and CES. The time spent for bouting in

the baits containing 2% ESP and EA was non-significantly more

than that spent in bait containing 2% CES.

In overall, no significant difference was found in

average food consumption (g/kg bw) for 2 hr a day between male

and female rats in respect of WSO bait without additive as

well as of WSO bait containing 2% ESP, EA and CES. The number

of bouts made towards WSO bait without additive were

significantly (P ≤ 0.05) more by male rats compared to female

rats. However, there was no significant difference in number

of bouts made towards WSO bait containing three bait additives

between male and female rats. The time spent in bouting

towards WSO bait without additive was also significantly (P ≤

0.05) more by male rats compared to female rats and there was

no significant difference in time spent in bouting towards WSO

bait containing three bait additives between male and female

rats.

These results in no-choice indicate almost similar

consumptions of WSO bait without additive and those with

additives. Among baits with different additives, however, the

bait consumption, number of bouts made and the time spent were

non-significantly more towards the bait containing 2% EA than

baits containing 2% ESP and CES by female rats. The results on

acceptance of baits by male rats are thus inconclusive. The

results of no-choice feeding tests thus indicate sex specific

76

`

differences in behaviour of R. rattus towards WSO bait containing

poultry egg components as bait additives.

Treatment of three blocks (I, II and III) of poultry farm

at village Ghutani Kalan, District Ludhiana with 2% zinc

phosphide bait without additive and that containing 2% EA and

ESP, respectively revealed 26.95, 68.20 and 68.65% consumption

of treated bait in blocks I, II and III, respectively.

Consumption of treated baits was significantly (P ≤ 0.05) high

in blocks II and III as compared to that in block I. No

significant difference was found in pre-census bait

consumption among the four treated and untreated blocks

indicating almost similar level of rodent activity in all the

blocks. Percent reduction in rodent activity in blocks treated

with 2% zinc phosphide without additive and those containing

2% EA (block II) and 2% ESP (block III) was found reduced by

33.52, 28.13 and 44.41%, respectively with respect to

untreated block and by 24.07, 19.23 and 42.69%, respectively

with respect to same block. However, there was no significant

difference in reduction in rodent activity between the blocks

I, II and III. In block I, significantly less acceptance of 2%

zinc phosphide without additive may be due to neophobia

behaviour shown by rats towards highly toxic rodenticide

leading to subsequent bait aversion. The results suggest the

potential of 2% EA and ESP in increasing the acceptance of 2%

zinc phosphide bait.

Rodent trapping with wooden single rat traps was carried

out in sheds of poultry farm located in the campus of GADVASU,

Ludhiana. Each trap was baited with a piece of chapatti

smeared by weight with 2% ESP and EA. For the first time,

trapping was carried out from all the sheds by alternatively

77

`

placing traps containing chapatti pieces without additive and

those containing chapatti pieces smeared with 2% ESP. Results

revealed total trap index of R. rattus to be 11.60 in traps

baited with ESP and 13.38 in traps baited with chapatti piece

without any additive. No significant difference was found in

total trap index as well as that of male and female rats

between the traps baited with chapatti pieces with and without

ESP. Also no significant difference was found in the trap

index between the two sexes.

For the second time, trapping was carried out by

alternatively placing traps containing chapatti pieces without

additive and those containing chapatti pieces smeared with 2%

EA. Results revealed total trap index of R. rattus to be 5.35 in

traps baited with EA and 7.13 in traps baited with chapatti

piece without any additive. No significant difference was

found in total trap index as well as that of male and female

rats between the traps baited with chapatti pieces with and

without egg albumin. Also no significant difference was found

in the trap index between the two sexes.

For the third time, trapping was carried out from all the

sheds by alternatively placing traps containing chapatti

pieces smeared with 2% ESP and 2% EA. Results revealed total

trap index of R. rattus to be 13.69 in traps baited with ESP and

12.79 in traps baited with EA. No significant difference was

found in total trap index as well as that of male and female

rats between the traps baited with chapatti pieces smeared

with ESP and EA. Also no significant difference was found in

the trap index of traps between the two sexes.

In overall, the results of poultry trapping experiments

revealed no significant effect of poultry egg components i.e.

78

`

ESP and EA in trapping of rats by single catch rat traps. This

may be due to the reason that rats might not have come to know

about the additive smeared on chapatti piece before they

pulled it from the string of the trap and got trapped.

During present studies, results of laboratory experiments

in bi-choice revealed significantly (P ≤ 0.05) higher

consumption of WSO bait containing all the three

concentrations of EA and 2 and 5% ESP by female rats and that

of bait containing 5 and 10% EA by male rats. In no-choice

experiments, the consumption, number of bouts made and the

time spent in WSO bait containing 2% EA by female rats was

non-significantly high than that containing 2% ESP and CES. In

poultry farm, significantly higher consumption of 2% zinc

phosphide bait containing 2% EA and 2% ESP leading to upto

44.41% reduction in rodent activity was observed compared to

33.52% reduction in rodent activity with only 2% zinc

phosphide treatment.

We may therefore assume that use of 2% egg albumin and

egg shell powder in cereal based bait may enhance bait intake

as compared to plain bait. This combination may yield

significant control of R. rattus if used in 2% zinc phosphide

bait.

79

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ENHANCING POISON BAIT ACCEPTANCE AND TRAPPING OF Rattus rattus USING POULTRY EGG COMPONENTS AS BAIT...

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