Reproductive seasonality in buffalo

(Bubalus bubalis)

Luigi Zicarelli
zicarell@unina.it

Dipartimento di Medicina Veterinaria e

Produzioni Animali
Universit Federico II
Via Delpino, 1 80137 Napoli, Italy
Introduction
 data of FAO
cattle buffalo
n n
n. countries in which is bred 212 43
% 20
it is likely that buffalo is bred 212 129
(Misra AK & Tyagi S 2007)
% 61
 buffalo and cattle population trend (%)
between 1961 and 2008
cattle Buffalo
world % %
between 1961 and 2013 59 125
between 1961 and 2013 (x 106) 1494/942 199/89
between 1990 and 2013 15 35 (*)
between 1990 and 2013 (x 106) 1494/1297 199/148
Countries where two species are bred
between 1990 and 2013 29 35 (*)
between 1990 and 2013 (x 106) 705/547 199/148
(*) This increase is mainly due to the increase of River buffalo (50
n; milk production), while the Swamp (48 n; draught animal
power), decreased of 27% (L. C. Cruz, 2010).
 the buffalo population increase in the countries with
high temperatures and high humidity rate
Dairy cow (*) is not able to totally exploit its
genetic merit
River buffalo (**) is able to totally exploit its genetic
merit
(*) the number of bovine population is also due to the
diminution of draught animals, but it is confirmed that the
buffalo is the ruminant for milk production in these zones

(**) if nutritive requirements are satisfied, the production of

buffaloes in Tropical zones and those in Temperate Areas are
very similar
 Why the buffalo population increase in Southern Asia if the
milk production increase more in cattle?
factors that reduce the advantage deriving by the cattle buffalo
higher production/head
susceptibility to the disease + -
adaptability to the tropical environment - +
herd life (*) - +
culling rate (*) + -
availability of subjects for beef production - +
coincidence between production and forage - +
availability
(*) it is necessary to breed around 90 and 42 subjects for 100 heads/year,
in bovine and buffalo, respectively. Consequently, the same number of
bovines will need a larger agriculture area compared to buffaloes, in
Countries where the farms are characterized by low dimensions.
Importance of buffalo milk in
India, Nepal, Egypt and
Pakistan
 Consumption percentage (%) of buffalo milk:
buffalo milk is user more than cattle
80
75
70
65
60
%

55
50
45
40
1961 1970 1980 1990 2000 2012
India 55,88 56,70 56,70 56,64 56,85 55,00
Nepal 67,46 69,57 72,46 70,24 69,24 71,10
Egypt 66,05 63,73 65,79 56,21 55,34 44,92
Pakistan 71,40 71,30 74,46 75,16 67,78 63,85
 Consumption (calories) percentage (%) of buffalo
milk
85
80
75
70
65
%

60
55
50
45
40
1961 1970 1980 1990 2000 2012
India 65,97 66,72 66,72 66,67 66,85 65,17
Nepal 76,04 77,77 80,11 78,32 77,51 79,02
Egypt 74,87 72,90 74,64 66,27 65,48 55,52
Pakistan 79,26 79,18 81,70 82,25 76,30 73,00
Importance of buffalo milk in
Iran, Iraq and Italy
 Consumption percentage (%) of buffalo milk (Iran,
Iraq, Italy)
25

20

15
%

10

0
1961 1970 1980 1990 2000 2012
Iran 9,25 5,77 4,01 4,46 4,34 2,17
Iraq 11,76 9,50 8,73 7,77 21,52 10,82
Italy 0,14 0,40 0,63 0,39 1,09 1,79
 Consumption (calories) percentage (%) of
buffalo milk
35

30

25

20
%

15

10

0
1961 1970 1980 1990 2000 2012
Iran 13,50 8,57 6,01 6,67 6,50 3,28
Iraq 16,95 13,84 12,77 11,42 29,57 15,66
Italy 0,22 0,61 0,95 0,59 1,65 2,71
 Consumption percentage (%) of quantity (kg)
or kcal of buffalo milk in countries with more
(a) or less (b) production
80
70
60
50
40
%

30
20
10
0
1961 1970 1980 1990 2000 2012
kg (a) 65,20 65,32 67,35 64,56 62,30 58,72
kcal (a) 74,04 74,14 75,79 73,38 71,54 68,18
kg (b) 7,05 5,22 4,45 4,21 8,98 4,92
kcal (b) 10,22 7,67 6,58 6,23 12,57 7,22
 Aim of the review
examine the main factors affecting
seasonality in buffalo and the strategies
that may be adopted to enhance the
reproductive performances
 The reproductive problems are different
depending on the area of the breeding
summer anoestrus

In tropical heat stress forage scarcity

countries North of
the equator
(TCNE)

in Italy the temperatures diet is constant

are milder throughout the
year
The main reproductive characteristics of buffalo (Madan,
1.988 ; Misra, 1.997; Misra and Tyagi, 2.007)

the delayed puberty

the prolonged post-partum ovarian
inactivity
the long inter-calving periods
the tendency toward seasonality
Delayed Puberty
Average age (month) at first calving between
1975 and 2005 in 30,753 primiparous buffaloes
bred in 86 farm in Italy (Zicarelli, 2007).
Percentage of animals that deliver at
different age (average = 30 months).
 rationing weaning age at first
calving
South Buffalo milk 7-8 months lower than
America + pasture old 36 months
Italy milk replace 100 days old 28-36
and TMR (*) months
developing Buffalo milk 3 4 months 40 months
countries + forage old
(*) rationing characterized by energy density of 0.8 MFU
and 12.5-13% protein content on dry matter
Reproductive seasonality
 Reproduction in function of the month gestation and
the light stimulus at North of Equator
sheep and goats horses and donkeys domestic buffalo in
tropical zones (31N
and 2S)
month gestation 5 11 and 12 month 10.5

reproduction Autumn Spring End of Summer -

takes place in Autumn
calving Spring Spring End of summer -
autumn
sensitive to decreasing of increasing of light decreasing of light
light stimulus stimulus stimulus
good availability Spring Spring (March- September - January
of forage at (February-May) May)
calving
Around the equatorial belt the reproductive season is highly conditioned
by forage availability (Vale 1988; Vale et al. 1990; 1996).
 Romanov
Russian breed of the Volga valley, females reach sexual
maturity at 3-4 months of age. Almost normal 4 live lambs,
able to reach the threshold of 6 - 7 live lambs.
It 'easy to change / extend the breeding season
Nutritional factors
The majority of authors of TCNE
attribute reproductive seasonality
to nutritional factors (Roy et al.
1968; Singh & Desai 1979; Shah
1990; Singh et al. 1988; Singh &
Lal 1992; Qureshi et al. 1999a).
 Out-of-breeding-mating-strategy
(OBMS)
It may be predicted that there is a tendency towards
seasonality as far away is the buffalo species from the
equator. Transferring the species to other breeding areas
has not modified hypophyseal hypothalamic axis
sensitivity to a decreasing light/dark ratio.

In Italy the seasonality makes necessary to change the

calving calendar in order to meet the milk market demand;
the OBMS entails the interruption of natural mating or the
use of AI between October and late January in adult
females (between September and late March in heifers),
which is the most favourable period for the
reproductive activity (Zicarelli , 1994; Zicarelli 1997b;
Campanile et al. 2009).
 Milk market demand (% M D) through the year, calving (%) and milk
production (%) in farms that utilize (% OBMS) or not (%No-OBMS) the OBMS
technique
 Melatonin and genetic factors

is possible in the buffalo

population identify subjects like
Soay or Romanov
Blood plasma levels of melatonin in female buffaloes which are readily ( Readily)
and laboriously ( -g- -Laboriously) driven out of breeding season, measured at
daylight (two hours before sunset - light), and two (2s), 4 (4s), 6 (6s) hours after
sunset, during the four season. Repeatability value of 0.733 (Di Palo et al., 1993).
Blood plasma melatonin secretory pattern at each two hour after sunset throughout
different months (B = buffaloes changed of farm)
Seasonality of calving in Nili-Ravi Buffaloes, purebred Sahiwal
and cross-bred cattle in Pakistan (Hassan et al., 2007)
 Factors affecting seasonality
nutritional factors
(Roy et al. 1968; Singh & Desai 1979; Shah 1990; Singh et al.
1988; Singh & Lal 1992; Qureshi et al. 1999a)
Seasonality in Italy is also found on farms where a constant
balanced diet is provided around year (Zicarelli 1994),
demonstrating that the Mediterranean Italian Buffalo is not
autochthonous.
heat stress
If heat stress was the main cause of the anoestrus it should be
admitted that it adversely affects the reproductive activity of
buffalo compared to Bos indicus.
In Italy exactly the opposite pattern is observed because in out-of-
season mated buffaloes the percentage of conception
increases between July and September, a period during which
the Holstein shows a low rate of conception.
Nutritional factors
The majority of authors of TCNE
attribute reproductive seasonality to
nutritional factors (Roy et al. 1968;
Singh & Desai 1979; Shah 1990;
Singh et al. 1988; Singh & Lal 1992;
Qureshi et al. 1999a).
 Reproductive efficiency
The different reproductive efficiency of buffalo species
compared to the cow is due above all to these
features:
1) in the buffalo is present a lower number of
primordial (Le Van et al. 1989) and antral follicles.
The number of oocyte in a buffalo calf is 1/5 of that
recorded in a bovine calf (Le Van et al. 1989;
Gasparrini 2002);
the seasonality depends on the low number of follicles
or is the low number of follicles that determines the
seasonality?
The buffalo is very big but the ovary
is very small
The buffalo is very big but the ovary
is very small

Some recent papers

(Lopez-Gautius et al. 2005)
report an incidence of
double ovulations of 15.5%
in dairy cows and similar
value has been previously
reported in buffalo (Zicarelli
et al. 1988).
In the latter, however, only
0.06% of double ovulation
lead to a pregnancy twin.
 The reproductive efficiency decreases when

a) need to modify the calving calendar in order

to meet the market demand.
b) After calving, due to the low number of
follicles and waves, few cycles occur and
hence, if conception does not take place, an
anoestrus of variable length begins (Zicarelli et
al. 1994).
 Reproductive efficiency and transition
period
With regard to this topic, the transition period and the post-
partum have a major importance for fertility in buffalo species
compared to the cow. It is worth to point out that in the last
two months of pregnancy the presence of

mycotoxins in the roughages

Clostridia
Coxiella burnetii
the incorrect input of Ca, P and crude proteins
often lead to the occurrence of vaginal or uterine
prolapse which impairs the R.R.C. (Zicarelli 2000).
 Delayed R.R.C. after calving
TCNE South of the equator over
the 20th parallel
bull is not present always present
(small farms)
the dry period of the scarcity of forage (November-March)
animals (March-June) abundance of forage
inter-calving intervals Prolonged (Shah 1990) less than 400 days
(Zicarelli 1994; Baruselli
2000)
However, unless the feed deficiencies are serious (Zicarelli 1999; 2001; 2004;
Paul & Lal 2010), nutritional factors in general do not play a significant role
because the species cannot yet be considered exactly as having a "lactiferus
habitus" and therefore a "catabolicus habitus" as the highly producing milk
bovine cow.
'temporary' and 'deep' anoestrus in OBMS buffaloes (C =

cyclic*; A= acyclic)

*R.R.C. within 70 days from calving

 Embryonic mortality and anestrus
After the observation of a corpus luteum, at the following clinical
examination the pregnancy is not always detected.

It is likely that this corpus luteum is characterised by an inadequate
luteinic secretion (short luteal phase or normal luteal phase but with
low progesterone production; (Zicarelli et al. 1997a; 1997b) like in
sheep upon the onset of anoestrus season and in bovine upon first
ovulation post-partum (Garverickh et al. 1992).

Often anoestrus condition preceded by an ovarian cycle

early embryonic mortality
 Embryonic death percentage in two farm in
function of month

The incidence of this event is considerably affected by the

farm factor and ranges between 10% and 45% if calculated
on the pregnancies after AI diagnosed by ultrasonography
at day 26 (Campanile et al. 2005)
 Reproductive efficiency

In Italy, the OBMS technique penalizes

fertility.
In fact, when the OBMS technique was not
applied, intercalving values of 400 410
days (Zicarelli et al. 2007b) have been
recorded in Italy. Only in a farm was found
a value of 445 days (Zicarelli et al. 1977).
 calving would conceive conception

between January between March and August - September

and March May longest inter-calving
When increasing period
day length
April - June between June show the shortest
August inter-calving period
When decreasing
day length
When the OBMS technique is not used, in autumn also the old
buffaloes (with more than 7 calving and still not pregnant after 7-9
months from last calving) spontaneously conceive. The BCS of
buffalo normally worsen after 12-15 years old and hence farmers
are not encouraged to eliminate the animals that are still not
pregnant at the end of lactation. This is one of the causes of the
prolongation of the inter-calving period (Zicarelli 2007b).
 Reproductive efficiency
Therefore, we can conclude that in Italy
nutrition and the temperature (especially if
a swimming-pool is present in the farm; Di
Palo et al. 2009; Neglia et al. 2009) are
not able to affect the inter-calving period.

The main factor at the latitude of 42-45, is

the light stimulus.
 The lower concentration of calving observed between
January and May depends on the reduced conception
rate between March and July.

This phenomenon is influenced by the hot and dry

climate (in accord to the majority of TCNE authors =
summer anoestrus) and the anoestrus of buffalo depend
on the heat stress (India, Pakistan) or to the low
environmental humidity (Venezuela).

In Italy, in farm that do not use the OBMS, a drop of

calving is also observed between March and June,
period of the year in which, unlike India, climate is either
cold or mild and moderately rainy.
 Monthly conception: Italy (Zicarelli et al., Monthly max temperature in
1977), India (Singh, 1988), Venezuele -
Guanota - farm, Apure State (Zicarelli, Naples (OBMS), Dheli,
1993), Pakistan (Hassan et al., 2007) Caracas and Lahore

The fertility markedly improves between August and September, period that in Italy
coincides with the highest temperature and THI of the year. This observation makes
buffalo much different from cattle.
 Milk market demand (% M D) through the year, calving (%) and milk
production (%) in farms that utilize (% OBMS) or not (%No-OBMS) the OBMS
technique
 Monthly conception: Italy (Zicarelli et al.,
1977), India (Singh, 1988), Venezuele - Monthly rainfall (mm)
Guanota - farm, Apure State (Zicarelli,
1993), Pakistan (Hassan et al., 2007)

The fertility markedly improves between August and September, period that in Italy
coincides with the highest temperature and THI of the year. This observation makes
buffalo much different from cattle.
 Monthly conception: Italy (Zicarelli et al., Trend of daily light hours in Naples,
1977), India (Singh, 1988), Venezuele - Dheli, Appure (Venezuele), Lahore
Guanota - farm, Apure State (Zicarelli,
1993), Pakistan (Hassan et al., 2007)

The trend of daily light hours, although with different daily values, is shared by all the
areas of breeding situated North of the equator
 Trend of daily light hours in Brazil in
Monthly calving percentage in Brazil in function of latitude ()
function of latitude ()
(Baruselli et al, 2000)

The trend of daily light hours, although with different daily values, is shared by the
different latitude of breeding situated South of the equator
Latitude

0 a 8

8 a 16

16 a 24

24 a 32
Monthly distribution in functione of distane from Equator (n = 16.487) in
Brazil, 2001
Baruselli et al. 2001

Cleavage rate in different seasons in
relation to day light hours
Percentages of tight morulae-
blastocysts (TM + BL) and good
quality blastocysts (BL G 1&2) in
different seasons in relation to day
light hours

Gasparrini et al., unpublished observations: 26,267 COC and 12,163 COC A+B
The main reproductive characteristics of buffalo (Madan,
1.988 ; Misra, 1.997; Misra and Tyagi, 2.007)

the prolonged post-partum

ovarian inactivity

the long inter-calving periods

the tendency toward seasonality
 Days of presence of the bull, % of presence/yare,
concep5on rate and days of intercalving
days of % days of bull
presence of presence % concep5on Intercalving
the bull (days/365d) rate (days)
Jan 270 73,97 85,71 464
FEB 240 65,75 86,59 427
MAR 210 57,53 53,21 428
APR 180 49,32 60,56 406
MAY 150 41,10 86,67 399
JUN 120 32,88 65,38 400
JUL 90 24,66 67,16 387
AUG 60 16,44 36,36 365
 Percentage of days where the bull is present (february
september) and intercalving (days)
470 intercalving
% days of bull presence
70 450

60 430

50 410
%

40 390

30 370

20 350

10
JUL
Jan
FEB
MAR
APR

JUN

AUG
MAY
 Fertility rate (%) and daily hours of light in Italy

Ligth hours (Italy)

Conception rate (%) of 16
pluriparous and primiparous
90
15
70
14
50
%

13
30

hours
12
10
ave
FE MA AP MA JU AU 11
Jan JUL rag
B R R Y N G
e
prim 89,4 77,0 70,2 65,2 56,5 47,2 38,1 64,3 10
plur 84 82,1 71,5 72,3 69,7 60,9 41 21,6 65,0 1 2 3 4 5 6 7 8 9 101112
 Conception rate (%) of pluriparous of 3 farm
100
90
80
70
60
50
%

40
30
20
10
0
avera
Jan FEB MAR APR MAY JUN JUL AUG
ge
sa % 91,1 87,6 73,4 74,7 81,4 61,6 55,0 29,8 69,3
st % 79,4 75,4 72,0 70,7 62,7 56,5 37,1 11,1 58,1
t% 80,5 83,9 68,0 71,1 62,6 66,4 25,8 25,0 60,4
 Conception rate (%) of primiparous of 3 farm
100

90

80

70

60

50
%

40

30

20

10

0
Jan FEB MAR APR MAY JUN JUL average
sa % 94,44 75,23 72,94 62,64 52,78 29,73 54,05 63,12
st % 92,86 75,00 76,39 60,71 55,56 50,00 0,00 58,65
t% 82,76 82,86 60,08 73,77 63,57 70,63 33,33 66,71
 Conception rate in pluriparous (PLUR) and in
primiparous (PRIM) of 3 farms
70,0

68,0

66,0

64,0
%

62,0

60,0

58,0

56,0
sa % st % t%
average PLUR 69,3 58,1 60,4
average PRIM 63,12 58,65 66,71
 Strategies to enhance reproductive
performance

a) Strategies to anticipate puberty

b) Treatments to reduce embryonic
mortality
c) treatments for anoestrus
d) improvement of welfare
 Puberty
The delayed puberty and the consequent older
age at first calving depend on both inappropriate
weaning techniques and inadequate feeding
during growth.
Age at first calving is 28-32 months (Zicarelli,
2007) with a rationing characterized by

energy density of 0.8 MFU (MUF = Milk unit forages

= 1700 kcal NEl)

12.5-13% protein content on dry matter

forage/concentrate ratio of 50-60%

 Embryonic mortality
Embryonic mortality in buffalo species is
primarily due to a reduced secretion of
progesterone by corpus luteum.
Unlike cattle, treatments on day 5 post-
insemination, do not have any effect in
reducing embryonic mortality in buffalo
(Campanile et al. 2007), while delaying
treatment on Day 26 after AI with GnRH
agonist, hCG or progesterone reduces
embryonic mortality (Campanile et al.
2008).
 treatments for anoestrus
The treatments for anoestrus are based on the
use of progesterone devices combined with
PMSG. However, the results are influenced by
the year/farm effect and therefore are variable.
The above treatments in natural mating
conditions do not have an immediate impact, but
especially in primiparous, they have a beneficial
effect on the R.R.C..
Unsatisfactory responses, in my opinion, are
perhaps more useful because they lead to
assess the environmental causes that determine
the failures (Zicarelli et al. 1994).
 Improvement of welfare:
swimming-pool presence
Although the light/dark ratio is the main factor affecting
reproductive efficiency, another important factor is the
satisfaction of the buffalo phisiological need of water for
bathing.
The swimming-pool presence reduced (Zicarelli 2002; Di
Palo et al. 2009; Neglia et al. 2009) the not pregnant
buffaloes/corpora lutea found at rectal examination ratio
(NP/CL) in buffaloes that calved between April and
August, because it acted clearly as a tool against heat
stress.
NP/CL ratio, as expression of anomalous oestrous cycle
or embryonic mortality, may be proposed as a specific
tool for evaluating buffalo welfare (Di Palo et al. 2009).
NP/CL ratio, as expression of anomalous oestrous cycle or embryonic mortality, may
be proposed as a specific tool for evaluating buffalo welfare (Di Palo et al. 2009).
 Improvement of welfare: space
We have demonstrated that increased
space and better welfare conditions
improve fertility in Italy.
These finding suggest that the effects of
season can in part be attenuated by
improving the welfare status of the
animals.
% of farms that resume cyclic activity % of farms that resume cyclic activity
in different months on June or July and use swimming
pools, pasture, increased space, covered
sheds (less light)
Calving calendar (%) of pluriparous buffalo cows in relation to the presence of
352 and 451 heads on the same space and daily light hours (h) at conception.
 Conclusions
The seasonality in buffalo species is influenced by the
light/dark ratio throughout the year.
In some Countries the seasonality is influenced by
nutritional factors.
Furthermore, the improvement of the welfare of the
subjects (swimming-pool, space, brightness of the stalls)
can increase the percentage of calving between March -
June.
Delayed treatment of buffaloes with GnRH agonist, hCG
or progesterone on Day 26 after AI can reduce the
embryonic mortality in the months in which daylight
hours increase.
A significant improvement in reproductive efficiency can
be achieved by increasing the culling rate from 10%
-14% to 25% -30%, and hence eliminating older subjects
and those with reproductive problems.
 Gracias por su atencin
Obrigado por sua ateno
Thanks for your kind attention
 North of the Equator
Photo species weather Forage Month Breeding Month
period availabili gestation season calving
ty
Nega ewe temperate spring 5 autumn spring
tive
Nega buffalo tropical after the 10,5 after after
tive rainy the the
season rainy rainy
season season
- -
autumn autumn
Nega buffalo temperate spring 10,5 autumn End of
tive summer
-
autumn
Posi mare temperate spring 11 spring spring
tive
Reproduction in function of the month gestation and the light stimulus at Nord of
Equator
temperate zones north of the tropical zones (31N
Equator and 2S)
good availability of forage Spring calving (March-May) September - January
month gestation 5
species sheep and goats
reproduction takes place in Autumn

sensitive to decreasing of light stimulus

calving Spring
month gestation 11 and 12 month 10.5
species horses and donkeys domestic buffalo
reproduction takes place in previous spring Previous Autumn-
Winter
sensitive to increasing of light stimulus decreasing of light
stimulus
calving Spring End of summer -
autumn
In the heifers the fertility is not penalised by the season (Zicarelli
1992; 1994; Avallone et al. 1994)

summer = hot weather

T3 T4

TRH

hyperprolactinemia
contrary to assertions by Madan (1988)
 In the calving calendar of the farms that are
adopting the OBMS technique, when the
primiparous are excluded from the survey, a
decrease in calving rate between March and
June is observed in Italy, both if OBMS is
applied or not, confirming that the increase of
daily light hours (April-June) or a number of daily
light hours >12 h, negatively interfere with
reproductive activity.
Calving percentage in farms that utilize (%OBMS) or
not (%No-OBMS) the OBMS technique (without
primiparous)
Intercalving period recorded in four provinces (n = 2654) in
OBMS farms; 1, 2, 3 = January, February, March; 4, 5, 6 =
April, May, June; 7, 8, 9 = July, August, September; 10,
11, 12 = October, November, December.
 The maximum daily temperatures that are recorded in Italy
and in Venezuela make us rule out a direct action of
environmental temperature on anoestrus whereas, limitedly to
India and Pakistan, it is not possible to exclude that heat
stress, even if it is not the main factor, contributes to
aggravate the summer anoestrus.
Furthermore, it is worth noting that the R.R.C. coincides
(August-September) with a monthly maximum temperature of
28C (August) and 25C (September) in Italy, 25C (August)
and 27C (September) in Venezuela, 34C(September) in
India and 36C in Pakistan.
 Conclusions

The seasonality in buffalo is influenced by the light/dark ratio

throughout the year. In somecountries the seasonality is
influenced by nutritional factors.
Furthermore, an improvement in the welfare of buffalo
(swimming pool, space, brightness of the stalls, etc.) can
increase the
percentage of calvings between March and June. Delayed
treatment of buffaloes with GnRH agonist, hCG or
progesterone on day 25 after AI can reduce embryonic
mortality in the months in which daylight hours increase.
A significant improvement in reproductive efficiency can be
achieved by increasing the culling rate from 10-14% to
25-30%, and hence eliminating older buffalo and those with
reproductive problems.
 Male seasonality

We have recently verified (Pelagalli et al. 2009) that in April only 23%,
31% and 29% of the bulls showed values higher than the average value
respectively of testosterone, diidrotestosterone and androstenedione.