Nosema Analysis in Honey Bees (in English)

the Oregon State honeybee lab presents

testing for Naima your sample must

contain at least 100 bees from the

colony of Interest store them in 70%

ethanol or in the freezer until

analysis the items you will need for

analysis will be your stored

sample

forceps

a mortar and

pestal a dish or

container a cutting utensil like a

scapel or micro

scissors a graduated cylinder measuring

up to 100 ml and deionized

water disposable

pipets and a hemocytometer with cover

slip from your sample take out exactly

100 bees with forceps and place in your

container sever the abdomen from the

thorax of each bee using a cutting

utensil and forceps place all abdomen

into the

mortar grind your 100 abdomen with the

pestal it's important that all abdomen

have been thoroughly crushed for a

homogenized

sample measure out 100 ml of deionized

water using your graduated cylinder if

your sample doesn't contain exactly 100

abdomen add 1 mL of water per each B of

your

sample the water will curve at the

surface when poured into the graduated

cylinder make sure to measure using the

bottom of that

curve pour water into the

mortar using your disposable pipet stir

and draw up part of the

sample Place One Drop onto one of the

Triangular spots in the counting chamber

of the hemus autometer carefully place a

cover slip over the counting chamber

turn on the microscope light and set the

power to 400 times then place the hemos

autometer on the microscope and begin to

put in

Focus once your sample is in Focus move

the slide around to find a set of

guidelines we want to find where the

horizontal and vertical guidelines cross

each other since we found the horizontal

guidelines first we will move left and

right to find the vertical

ones we located the upper right hand

corner of the cross-hatching grid

count the total number of nzima spores

within this cornered Square that's

outlined by tripl lined grid

Marks here are some highlighted nzima

spores become familiar with their

distinct shape and outline it may take

some practice to decipher spores from

other debris within the

sample if a NOA Spore Falls within the

Border grid lines do not count the ones

that lie within the left and bottom

border but do count the ones that lie

within the top and right border move the

slide around and count all spores that

lie within each Corner square and the

Very middle square of the grid record

the total number of

spores you will multiply the total

number of counted nazma spores by

50,000 this will give you the estimated

number of nzima spores per B within your

sample for example if you counted 50

spores your first sample contains 2.5

million nazima spores per individual

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