Beltsville Poultry Semen Extender. 7.

Comparison of Commercial Diluents

for Holding Turkey Semen Six Hours at 15 C1'2

A. F. GIESEN

Department of Poultry Science, Alabama Agricultural Experiment Station,

Auburn University, Alabama 36849

T . J . SEXTON

US Department of Agriculture, SEARS, Avian Physiology Laboratory, Beltsville, Maryland 20705

(Received for publication February 16, 1982)

ABSTRACT The ability of three commercial semen diluents (Minnesota, I.M.V.-French and
Universal) and the Beltsville Poultry Semen Extender (BPSE) to maintain the fertilizing capacity of
turkey semen held 6 hr at 15 C was examined. Hens were inseminated weekly for 20 wk and
hatchability of eggs was determined. Neither candling nor true fertility of semen diluted 1:1 in
BPSE, the Minnesota, or I.M.V.-French diluent and held for 6 hr differed significantly from each
other. Fertility of semen held 6 hr in BPSE or Universal diluent was lower than their respective
unstored controls. No difference in fertility was observed between semen held in Minnesota or
I.M.V.-French diluent and their unstored controls. Significantly higher hatch of fertile eggs and of
all eggs set was obtained from hens inseminated with semen diluted (0 hr) with BPSE (90 and 86%)
than all other diluents. Hatch of fertile eggs was higher from hens inseminated with semen held 6
hr in Universal (90%) and French (86%) extenders and lower for the Minnesota (78%) extender. No
difference was observed in hatch of fertile eggs between the BPSE (84%) and French (86%) ex-
tender. Hatchability of all eggs set was higher when hens were inseminated with semen held 6 hr in
the I.M:V.-French diluent (81%) when compared to hens inseminated with the Minnesota (75%) or
Universal diluent (75%). No differences in hatch of all eggs set were noted when BPSE (77%) and
the commercial extenders were compared.
(Key words.- diluents, turkey, storage, semen,fertility, hatchability)
1983 Poultry Science 62:379-381

INTRODUCTION
A n u m b e r of semen diluents commercially
available are used in t h e artificial insemination
(AI) of t u r k e y s . R e c e n t r e p o r t s by Lake ( 1 9 7 8 )
and S e x t o n and Giesen ( 1 9 8 2 ) indicated t h a t
satisfactory fertility ( > 9 0 % ) could be o b t a i n e d
with t u r k e y semen diluted in s y n t h e t i c m e d i a
and held 5 t o 6 hr before AI. However, few
investigations have been c o n d u c t e d t o de-
t e r m i n e if the commercially available t u r k e y
diluents aid in semen preservation. T h e purpose
'Scientific Article No. 12-820184 of the Alabama
Agricultural Experiment Station.
2
Mention of a trade name, proprietary product
or specific equipment does not constitute a guarantee
or warranty by the US Department of Agriculture
or Auburn University and does not imply its approval
to the exclusion of other products that may be sui-
table.
'Minnesota Turkey Growers Association, St.
Paul, MN 55114.
4
Applied Genetics Laboratory, Omaha, NE 68127.
5
1.M.V. International Corporation, L'Aigle, France.
of this e x p e r i m e n t was t o c o m p a r e t h e ability
of t h e Beltsville P o u l t r y Semen E x t e n d e r
(BPSE) and t h r e e commercial diluents, Min-
n e s o t a E x t e n d e r ( M T G A ) , 3 Universal M e d i u m
(Universal), 4 or I.M.V.-French Extender
( F r e n c h ) , 5 t o m a i n t a i n t u r k e y semen for 6 h r a t
15 C.
MATERIALS AND METHODS
Semen samples were collected from at least
10 Nicholas Large White males and p o o l e d
immediately after collection. Six cubic cen-
timeters of undiluted semen were divided into
four equal parts and diluted 1:1 with t h e
a p p r o p r i a t e e x t e n d e r within 30 m i n of col-
lection. T h e BPSE was prepared as outlined by
S e x t o n ( 1 9 8 0 ) , and t h e M T G A , F r e n c h , a n d
Universal diluents were purchased from com-
mercial sources. One and one-half cubic cen-
timeters of each of t h e diluted samples was
used immediately to inseminate the control
hens (0 hr). T h e remaining samples were placed
in four 10-ml E r l e n m e y e r flasks, covered
loosely with a l u m i n u m foil, and held at 15 C

379
380 GIESEN AND SEXTON
for 6 hr. During the holding period the samples
were constantly agitated on a Lab-Line Junior
Orbit Shaker at 150 rpm.
Eighty Nicholas Large White hens, housed
in floor pens, in their first-week of egg pro-
duction, were used with 10 hens randomly
assigned to each of the eight experimental
treatments. The hens were inseminated on 3
consecutive days during the first week of egg
production and then weekly for 19 weeks. Each
hen was inseminated with a dose of .1 ml
diluted semen containing approximately 365
million sperm. Sperm concentration was
determined for each undiluted sample with a
Klett-Summerson photometer at 400 to 465
nm (Cecil, 1981).
Eggs were collected daily, stored at 15 C,
and incubated within 7 days. Fertility and
hatchability were determined on approximately
862 eggs per treatment according to the pro-
cedures outlined by Sexton and Giesen (1982).
All fertility and hatchability data presented
as percentages were subjected to arcsin trans-
formation prior to analyses. Significance of
data was determined at the 5% level of prob-
ability by analysis of variance (ANOVA)
(Statistical Analysis System, 1979). Dif-
ferences between treatment means were
estimated by Duncan's multiple range test. With
a significant interaction between diluent and
holding time, a comparison of diluents within
holding times and a comparison of holding time
within diluents were accomplished by separate
ANOVA and Duncan's test. Standard errors
of the means were calculated from the variance
derived from the ANOVA.
TABLE 1. Fecundity of turkey semen held at 15 C for 0 or 6 hr in various diluents1
% Fertility
True Candled
Diluent 0 6 0 6
BPSE 96 a - A 9 1 b,B 94a,A
MTGA 93ab,C 96a.A 90ab,A
French 92 b > A 94ab,A 89 b > A
Universal 94ab,A 83CB 90ab,A
SEM 3 2
Means within columns followed by a different capital superscript letter are significantly different
(P<.05).
A,B X .
Means within rows within a parameter followed by a different lower case superscript letter are signi-
ficantly different (P<.05).
1
Mean and SEM of 862 eggs set per treatment.
RESULTS AND DISCUSSION
The data reported in Table 1 for hatchability
of all eggs set (HAS) exhibit clearly a difference
between semen treatments. The HAS was
highest (86%) in hens inseminated with semen
diluted in BPSE and used immediately. The
HAS was higher from hens inseminated with
semen held 6 hr in the French diluent (81%)
than for hens inseminated with semen stored in
either the MTGA (75%) or Universal (75%)
diluents. The HAS did not differ between the
French (81%) and the BPSE (77%) diluents.
The relationship of hatchability of fertile
eggs (HFE) to semen treatments was inter-
esting. For example, HFE from hens insem-
inated with semen held 6 hr in the Universal
diluent was higher than the 0 hr control. In
contrast, HFE from hens inseminated with
semen held 6 hr in either BPSE or the MTGA
diluent was lower than their respective 0 hr
controls. A possible explanation may be as-
sociated with the relationship of "stale sperm"
to embryonic death reported by Nalbandov and
Card (1943) and Dharmarajan (1950). Both
reports indicated that in vivo aging of chicken
sperm resulted in an increased incidence of
embryonic mortality and abnormal early
embryonic growth in the incubated egg. A
similar sperm aging effect in vitro caused by
holding semen at 15 C in the presence of
different extenders, could have influenced the
hatchability of fertile eggs in this study.
Holding semen for 6 hr significantly de-
pressed the fertilizing capacity (true and
candling) or semen diluted with the Universal
diluent and the BPSE when compared to the 0
% Hatchability
Fertile eggs All eggs set
0 6 0 6
88a.B 90a>B 84b,B 86a.A 77ab,C
92*,A 84b.A 78C.B 7 9 b,A 75b,A
9 1 a, A 84°.A 86ab.A 7 7 b,A 8ia,A
8lM 84b,B 90a.A 7 9 b,A 7 5 b,A
3 2
 BELTSVILLE POULTRY SEMEN EXTENDER
hr control, whereas the fertilizing capacity of
semen diluted with the MTGA or French
diluent did not change over the 6 hr holding
period. The high fertility levels (>90%) of
unstored semen (Table 1) are in agreement with
previous reports using BPSE (Sexton, 1977;
Sexton and Giesen, 1982), MTGA diluent
(Ogasawara and Ernst, 1970; Brown and
Graham, 1976; Hanson and Graham, 1976),
and the Universal diluent (Sexton, 1977).
ACKNOWLEDGMENTS
The authors wish to express appreciation to
D. M. Gavelek and V. W. Smoot for their
technical assistance.
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turkeys. Poultry Sci. 60:1049-1055.
381
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