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BREEDING FOR SUSTAINABILITY: HOW REPRODUCTIVE
BIOTECHNOLOGIES CAN HELP BUFFALO FARMERS COMBAT
CLIMATE CHANGE
Reproduciendo para la sostenibilidad: cómo las biotecnologías reproductivas pueden
ayudar a los criadores de búfalos a combatir el cambio climático
Pietro Sampaio Baruselli1*, Laís Ângelo de Abreu1, Vanessa Romário de Paula2, Sofía Albertini¹,
Guilherme Felipe Ferreira dos Santos¹, Lígia Mattos Rebeis¹, Emanuelle Almeida Gricio¹, Nelcio A.T. de Carvalho3,
Otavio Bernardes4
1 Department of Animal Reproduction, Faculty of Veterinary Medicine and Animal Science, University of São Paulo,
São Paulo, SP, Brazil
2Instituto Paulista de Ensino e Pesquisa, Empresa Brasileira de Pesquisa Agropecuária – EMBRAPA, Juiz de Fora, MG, Brasi
3Research and Development Unit of Registro / Diversied Animal Science Research Center
4Fazenda Paineiras da Ingaí, Sarapuí, SP, Brazil
*Corresponding author: Baruselli, Pietro Sampaio (barusell@usp.br)
ABSTRACT
The global attention on enteric CH4 production in ruminants
requires a response that involves collaboration between re-
searchers and industry. Future generations of bu󰀨aloes will be
characterized by better e󰀩ciency and fertility, which may re-
duce CH4 emission intensity. This goal will result from balanced
multi-trait selection and the introduction of e󰀩cient reproductive
and productive management. Currently, e󰀩cient reproductive
programs using assisted reproductive technologies (ARTs)
are available on bu󰀨alo farms. Our expanding knowledge of
ovarian function during the bu󰀨alo estrous cycle has given new
approaches for precisely synchronizing follicular development
and ovulation to apply ARTs consistently. Synchronization pro-
tocols are designed to control both luteal and follicular function
and permit xed-time AI with high pregnancy rates during the
breeding (autumn-winter) and non-breeding (spring-summer)
seasons. Additionally, it allows the initiation of superstimulatory
treatments at a self-appointed time, providing opportunities to
superstimulate bu󰀨aloe donors associated with ovum pick-up
(OPU) and in vitro embryo production (IVEP). Furthermore, it
allows xed-time embryo transfer in recipients, with high ef-
ciency and no need for estrus detection. Thus, ARTs, such
as AI and ET, are applied for bu󰀨alo’s targeted multiplication
and dispersal with dened production and environmental cre-
dentials. Also, the urgency in moving to the next generation
of bu󰀨aloes will increase the production of embryos from ge-
nomically dened prepubertal heifers. Using these biotechnol-
ogies will reduce generation interval and accelerate the rate
of genetic improvement to bu󰀨alo, dened by better e󰀩ciency
and fertility and lower CH4 emission. The challenge remains to
communicate the importance of bu󰀨aloes for food security and
the environment.
Keywords: enteric methane, e󰀩ciency, fertility, assisted repro-
ductive technology.
RESUMEN
La atención mundial sobre la producción de CH4 entérico en
rumiantes requiere una respuesta que implique la colaboración
entre investigadores y la industria. Las generaciones futuras de
búfalos se caracterizarán por una mayor eciencia y fertilidad,
lo que puede reducir la intensidad de las emisiones de CH4.
Este objetivo será el resultado de una selección equilibrada
de múltiples rasgos y la introducción de un manejo reproducti-
vo y productivo eciente. Actualmente, las granjas de búfalos
cuentan con programas reproductivos ecientes que utilizan
tecnologías de reproducción asistida (ART). Nuestro creciente
conocimiento sobre la función ovárica durante el ciclo estral
de las búfalas ha brindado nuevos enfoques para sincronizar
con precisión el desarrollo folicular y la ovulación para aplicar
las ART de manera consistente. Los protocolos de sincroniza-
ción están diseñados para controlar la función lútea y folicular
y permitir la IA a tiempo jo (IATF) con altas tasas de preñez
durante las temporadas de reproducción (otoño-invierno) y no
reproductiva (primavera-verano). Además, permite el inicio de
tratamientos de superestimulación en el momento que usted
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elija, brindando oportunidades para superestimular a los do-
nantes de búfalas asociados con la recogida de óvulos (OPU) y
la producción de embriones in vitro (IVEP). Además, permite la
transferencia de embriones a tiempo jo (TETF) en las recep-
toras, con alta eciencia y sin necesidad de detección de estro.
Por lo tanto, las ART, como la IA y la ET, se aplican para la mul-
tiplicación y dispersión selectiva del búfalo con credenciales
ambientales y de producción denidas. Además, la urgencia
de pasar a la próxima generación de búfalos aumentará la pro-
ducción de embriones a partir de novillas prepúberes genómi-
camente denidas. El uso de estas biotecnologías reducirá el
intervalo generacional y acelerará la tasa de mejora genética
del búfalo, denida por una mayor eciencia y fertilidad y una
menor emisión de CH4. El desafío sigue siendo comunicar la
importancia de los búfalos para la seguridad alimentaria y el
medio ambiente.
Palabras clave: metano entérico, eciencia, fertilidad, tecno-
logía de reproducción asistida.
INTRODUCTION
The world´s population is projected to increase by 24%
by 2050, potentially reaching 9.7 billion people [1]. Food pro-
duction must increase by 49% to sustain this population explo-
sion [2]. In this scenario, urbanization, and growing concerns
about the environmental impact of livestock farming demand
a long-term global strategy for more sustainable ruminant pro-
duction. Bu󰀨alo, therefore, will continue to have a signicant
role in future global food security. The global bu󰀨alo population
is approximately 202 million head [3], compared to 1.5 billion
cattle [4].
Bu󰀨alo milk and meat products can meet human needs
for high-quality protein. They excel over cattle exploiting
low-quality feed typical of many rearing areas and demonstrate
great adaptability to various management and temperature
conditions [5]. Furthermore, most bu󰀨alo production is carried
out extensively in pastures and savannas suited for low-input
and low-cost animal production. In South Asia, the River bu󰀨a-
lo is a primary source of milk and meat and has a crucial role
in food security. The riverine bu󰀨alo also supports high-value,
di󰀨erentiated food production in Europe and the Americas. The
Swamp bu󰀨alo is a vital draft animal and a source of food in
Southeast Asia and East Asia.
However, the environmental impact of ruminant produc-
tion has gained signicant attention worldwide [6, 7, 8]. Cattle
contribute around 4.5-5.0% of global anthropogenic methane
[9]. Enteric fermentation, with an annual emission of 87-97 Tg
(i.e., 1012g), is one of the agricultural sector’s signicant meth-
ane sources [10]. The global contribution of cattle and bu󰀨alo
to annual enteric methane emissions is 77% and 13%, respec-
tively [11]. The primary source of methane in ruminants orig-
inates from the enteric fermentation process, where complex
carbohydrates are converted into simple sugars by methano-
genic protozoa [12]. Extensive reviews have comprehensively
covered the biology and function of the rumen [13, 14]. The
quantity of methane an animal produces is signicantly inu-
enced by the relative abundance of ruminal methanogenic and
non-methanogenic microbes [12]. Microbial gene abundance
analysis advancements allow for determining ruminal microbe
populations [15]. In addition to enteric methane (CH4) produced
by the rumen, beef, and dairy production also contributes car-
bon dioxide (CO2; feed), nitrous oxide (N2O; feed production,
manure), and other CH4 (manure) to the total greenhouse gas
(GHG) budget of the production systems.
Malik et al. [16] compared the enteric methane yield be-
tween cattle and bu󰀨aloes under the same nutritional manage-
ment. Enteric methane emissions (g/d) depended on dry mat-
ter intake (kg/d). However, the methane yield (g/kg dry matter
intake; DMI) did not di󰀨er between species when fed on the
same diet (Cattle=13.4 g/kg DMI vs. Bu󰀨aloes=13.5 g/kg DMI).
This result conrms that methane yield depends on the diet
rather than the species compared. Thus, methane mitigation
strategies developed in one of the species can be e󰀨ective in
the other.
In this scenario, the use of assisted reproductive technol-
ogies can have a signicant impact on improving e󰀩ciency in
bu󰀨alo production systems. Reproductive technology has been
progressively rened in bu󰀨aloes, and today, the success of
articial insemination and embryo transfer is comparable to cat-
tle. Articial insemination (AI), combined with estrus synchroni-
zation, is a potent strategy of assisted reproduction technology
to improve reproductive e󰀩ciency and expedite genetic gain
in bu󰀨aloes [16]. Furthermore, embryo transfer (ET) enables
the multiplication of high maternal and paternal genetic value,
playing a more signicant role in the genetic enhancement of
this species [17]. This review seeks to demonstrate how assist-
ed reproductive technologies (ARTs) can improve reproductive
e󰀩ciency and harvest the next generation of bu󰀨aloes that pro-
duce more milk and meat to combat climate change.
REPRODUCTIVE EFFICIENCY IN BUFFALO AND
APPLICATION OF ARTIFICIAL INSEMINATION TO
IMPROVE PRODUCTION AND REDUCE METHANE
EMISSION
The cow-calf operation system utilizes approximately
70% of resources. Therefore, selection for reproductive ef-
ciency signicantly a󰀨ects farm e󰀩ciency, protability, and
sustainability. With high reproductive e󰀩ciency, fewer cows are
required to produce the next generation of calves, reducing re-
source requirements, herd methane production, and costs [18].
Furthermore, assisted reproduction technologies can also be
used to manipulate reproduction in bu󰀨alo. This includes syn-
chronization of the breeding time, inuencing the age at rst
breeding, the interval between the calving, and improving the
breeding during seasonal anestrus [16].
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13th World Bu󰀨alo Congress ~ 13er Congreso Mundial de Búfalos / Lectures / Reproduction ______________________________________
Articial insemination can be incorporated into bu󰀨alo
breeding programs to further improve reproductive e󰀩ciency
and genetic gain, collaborating to reduce CO2-eq emission in-
tensity. However, the traditional AI program e󰀩ciency needs to
be improved by low estrous detection. Bu󰀨alo presents a poor
manifestation of estrus symptoms, implying operational di󰀩cul-
ties in detecting estrus [19].
Furthermore, the success of reproductive programs is
closely related to the bu󰀨alo reproductive seasonality. Bu󰀨alo is
a seasonal reproductive species and becomes sexually active
in response to a decreasing day length (short days) in late sum-
mer to early autumn [20, 21]. During the non-breeding season,
bu󰀨alo often exhibit anestrus, which extends the anovulatory
period and reduces reproductive performance [22].
Nowadays, timed articial insemination (TAI) can be ap-
plied routinely in farm reproductive programs. TAI protocols are
designed to control both luteal and follicular function, permitting
the AI without estrus detection and during the anestrous peri-
od with high reproductive e󰀩ciency during the breeding and
non-breeding season [19, 23, 24]. Several studies demonstrate
that it is possible to establish an e󰀨ective AI program in bu󰀨a-
loes throughout the year, collaborating to increase the number
of pregnant bu󰀨aloes during the non-breeding season and dis-
tributing calving and milk production throughout the year. Using
reproductive programs with TAI followed by resynchronization,
it is possible to obtain high reproductive e󰀩ciency (>80% preg-
nancy rate after 3 FTAI) with inter-calving intervals close to 12
months (FIG. 1; adapted from Baruselli et al. [25]).
The e󰀩ciency of TAI in bu󰀨alo demonstrates that it is
possible to introduce e󰀩cient articial insemination programs
on farms that collaborate to increase the reproductive and ge-
netic e󰀩ciency of the herds.
EMBRYO TECHNOLOGY TO MITIGATE METHANE
EMISSION
In vivo, (superovulation; SOV) and in vitro (ovum pick-up
and in vitro embryo production; OPU/IVEP) embryo produc-
tions are reproductive biotechnologies used worldwide in beef
and dairy operations to disseminate the genetic material of su-
perior animals. Selection and genetic gain are essential to im-
prove e󰀩ciency, product quality, and sustainability [16]. When
comparing both biotechnologies in bu󰀨alo, OPU/IVEP demon-
strates higher e󰀩ciency and greater commercial applicability
than SOV. However, there are some limitations to using OPU/
IVEP, such as seasonality, the low number of antral follicles,
and the low quantity and quality of the recovered oocytes [17].
Experiments have been conducted to enhance OPU/
IVEP e󰀩ciency. In one study, Sá Filho et al. [26] demonstrated
FIGURE 1. Conception rate (P/AI) of lactating bu󰀨alo (n=510) submitted to xed time AI following resynchronization in
non-pregnant cows. Ultrasonography evaluation was performed to detect non-pregnant bu󰀨alos 30 days after AI for re-
synchronization. Pregnant bu󰀨aloes from the 1st FTAI with 50 days postpartum presented a 50% conception rate (CR) and
11.7 months of inter-calving interval (ICI). Pregnant bu󰀨aloes from the 2nd FTAI with 90 days postpartum presented a 43%
CR and 12.9 months of ICI. Pregnant bu󰀨aloes from the 3rd FTAI with 130 days postpartum presented a 30% CR and 14.3
months of ICI. After 3 FTAI, bu󰀨aloes presented an 80.1% pregnancy rate with a mean of 12.3 months of ICI (adapted from
Baruselli et al., 2003)
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that using bST increased the number of small antral follicles at
OPU. Additionally, bST tended to increase the number of re-
covered oocytes and improved the percentage of high-quali-
ty oocytes. However, bST showed no e󰀨ect on cleavage and
blastocyst production rates. In another study, Carvalho et al.
[27] showed that FSH treatment for superstimulation before as-
piration improved the outcomes of OPU/IVEP. FSH treatment
increased the proportion of large and medium follicles at OPU
and enhanced the viable oocyte rate, blastocyst rate, and num-
ber of embryos produced per OPU session.
The use of OPU/IVEP in females before puberty, apart
from the genetic gain inherent in this biotechnology, also re-
duces the generation intervals, further accelerating genetic
improvement. This technology can be employed in prepuberal
bu󰀨alo heifers, where ovaries have established follicular waves
and respond to superstimulation, or in bu󰀨alo calves, where
OPU is performed via laparoscopy [LOPU; 28, 29, 30]. LOPU
permits the recovery of oocytes from calves of two months of
age and the in vitro production of embryos that will be trans-
ferred to recipients. This technology allows a donor animal to
produce o󰀨spring before it reaches sexual maturity. The use of
young donors has two main key point that makes this alterna-
tive interesting: the rst one is the larger follicular population,
and the number of cumulus-oocyte complexes (COCs) recov-
ered, and the second is the shorter generation interval, increas-
ing genetic gain [30, 31].
In a study conducted by our group, we compared embryo
production in bu󰀨alo calves (2-4 months of age), prepubertal
bu󰀨alo heifers (13-15 months of age), and lactating bu󰀨alo
cows [28]. The treatment for calves involved using a sheep in-
travaginal P4 device on day 0 of the protocol, and for stimulat-
ing follicular growth, 140 mg of FSH was administered in four
decreasing doses every 12 hours on days 5 and 6. On day 7,
oocytes were recovered by LOPU in calves and through OPU
on a random day of the estrous cycle in prepubertal heifers
and lactating cows. The results showed that calves had a low-
er blastocyst production rate, but the number of embryos pro-
duced was similar between calves and lactating cows. Embryos
produced from calves (n=8) resulted in three pregnancies (3/8;
38%), which led to the birth of three healthy calves [28]. This
study demonstrated the feasibility of IVEP in young animals to
reduce generation interval and signicantly accelerate genetic
progress in bu󰀨aloes. However, calves were less e󰀩cient in
embryo production than prepubertal heifers and cows, and fur-
ther research is needed to optimize IVEP in young bu󰀨alo [30].
Regarding the impact of assisted reproductive tech-
niques on methane emissions in cattle operations, IVEP of oo-
cytes retrieved from young animals presents a viable approach
to achieving genetic gain and reducing generation intervals [8].
Although the e󰀩ciency of IVEP in young animals is relatively
lower due to hormonal and metabolic di󰀨erences, its integration
with genomic selection o󰀨ers a powerful strategy to enhance
genetic gain, e󰀩ciency, and fertility, as well as mitigate meth-
ane emissions in bu󰀨alo operations [32].
BALANCING FEED EFFICIENCY IN MEAT AND
MILK PRODUCTION WITH FERTILITY AND LOW
CO2-EQ EMISSION
Ruminants are crucial in maintaining sustainable agri-
cultural systems due to their distinctive capacity to transform
forages into high-quality meat and dairy products [33]. The link
between feed e󰀩ciency, methane production, and sustainabili-
ty has been known for over 20 years [34, 35, 5]. The relatively
high heritability of growth and feed e󰀩ciency in cattle was rec-
ognized some 70 years ago and subsequently conrmed [6,
36, 37, 38].
Furthermore, in tropical and subtropical regions, the
conjunction of elevated temperatures and humidity during the
summer months leads to the onset of reproductive problems,
decreasing milk and meat production in bu󰀨aloes [39, 40, 41,
42]. Implementing management techniques, such as active
cooling, is imperative to alleviate these stressors and uphold
a certain level of productivity. Additionally, the summer season
decreases feed quantity and quality, compounding the nutri-
tional challenges that impact reproductive capabilities [39, 42].
Beyond photoperiod, it is essential to address external inuenc-
es that detrimentally a󰀨ect reproduction and production to fully
capitalize on the potential a󰀨orded by the worldwide demand
for bu󰀨alo food items. Methods encompass targeted nutritional
enrichment, assisted reproductive technologies (ARTs) appli-
cation, and managerial tactics (such as cooling techniques and
ample resting areas) to enhance bu󰀨alo welfare within naturally
endowed and non-endowed production setups.
Bu󰀨alo farming has transitioned to a more intensive mod-
el, utilizing a feeding system structured around three distinct
rations corresponding to the primary bu󰀨alo production stages:
lactating cows, dry cows, and growing heifers. Their diet pri-
marily comprises maize silage and ryegrass hay, with addition-
al concentrates reserved solely for lactating bu󰀨alo cows [43].
These farming conditions developed for bu󰀨alo production in
Italy entail the absence of pasture access and wallowing water.
Recent studies suggest that incorporating more digest-
ible forages into ruminant diets may mitigate CO2 emissions,
even within intensive systems [44]. Despite this, the cumula-
tive emissions of free-ranging (FR) animals exceeded those of
conned (C) systems by approximately 662 kg CO2-eq. This
discrepancy stemmed from the animals in the FR system con-
suming a greater volume of brous feed than the C heifers. At
puberty, the heifers reached a weight of 402±3 and 382±3 kg in
systems C and FR, respectively. Di󰀨erences between groups
were signicant (p<0.05) due to the higher feeding regimen of
group C, the higher physical activity performed while grazing by
group FR, and the lower environmental temperature of the hilly
area where this group was located.
Nevertheless, these animals reached puberty at an age
not signicantly di󰀨erent from that observed in group C (p>0.05;
[45]). This nding has been attributed to the fact that grazing
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13th World Bu󰀨alo Congress ~ 13er Congreso Mundial de Búfalos / Lectures / Reproduction ______________________________________
animals used the available resources (pasture and feeding
supplementation) e󰀩ciently. In contrast, conned heifers used
spare nutrients only to increase their body mass after fullling
their requirements for development [46].
In temperate regions, bu󰀨alo experience a distinct sea-
sonal reproductive pattern inuenced by photoperiod and mel-
atonin secretion, as indicated by previous research [20, 47,
21]. Optimal conditions lead to a resumption of anoestrus in
bu󰀨alo within 30–90 days postpartum. However, factors includ-
ing inadequate nutrition and poor body condition [48], suckling
management [49], and climate [50] can signicantly delay this
process. For instance, bu󰀨alo in Sri Lanka under free grazing
with limited calf access to dams for suckling once a day re-
sumed estrous cycles within 30–60 days, whereas those ex-
posed to harsher conditions and free calf suckling remained in
anestrus for 150–200 days [51]. Bu󰀨alo’s postpartum LH se-
cretion remains low initially, with detectable episodic pulses a
few weeks before ovarian activity starts. Improved nutrition and
controlled suckling prompt LH release earlier than those with
poor nutrition or free suckling [52, 53]. There are recommended
methods to overcome extended postpartum anestrus in bu󰀨a-
lo, including ensuring proper nutrition before and after calving,
regulating calf suckling, and alleviating heat stress through ac-
tivities like wallowing or using water sprinklers [54], improving
the reproductive and productive e󰀩ciency.
Limited research has been conducted on evaluating the
environmental repercussions of dairy bu󰀨alo farms on environ-
mental sustainability. In a study, Pirlo et al. [55] found that the
ecological footprint of dairy bu󰀨alo farms, quantied in terms of
global warming potential, amounted to 5.07 kg of CO2 equiva-
lent per 1 kg of standardized bu󰀨alo milk. This gure is nearly
vefold greater than that generated by dairy cow farms [56].
This disparity could be attributed to the similarity in energy in-
puts and raw material acquisition between dairy bu󰀨alo and
cow farms, coupled with comparatively lower milk production
from bu󰀨alo.
According to Chirone et al. [57], bu󰀨aloes’ milk produc-
tivity varies from farm to farm and is a key factor determining
environmental performance. The remaining di󰀨erences are
explained by a combination of the type of feed (including the
portion cultivated in-house and purchased) and the strategy for
managing manure. These ndings reinforce the importance of
increasing the genetic capacity of bu󰀨aloes to produce milk and
meat more e󰀩ciently.
Bu󰀨aloes exhibit notable feed conversion e󰀩ciency and
sustain productivity even when subjected to diets limiting for
cattle [58]. In a previous investigation, metabolic condition and
reproductive performance were observed in Murrah bu󰀨alo
heifers fed either a high-energy (HE) or low-energy (LE) diet
[58]. Heifers following the HE diet displayed elevated plasma
levels of insulin, leptin, and T3, along with increased concen-
trations of IGF-1 in follicular uid and a higher oocyte quality
index. These outcomes highlight the positive e󰀨ect of the nu-
trition improving the reproduction performance and production
in bu󰀨aloes.
Recently, the currently used methods of estimating the
carbon footprint of processed animal products and dairy prod-
ucts should consider the subtraction of carbon emissions and
sequestration. According to De Vivo et al. [59], considering car-
bon sequestration and implementing this calculation method
would demonstrate sustainability regarding the carbon footprint
of agricultural products of animal origin, such as bu󰀨alo dairy
products (Mozzarella cheese) and meat products.
ENTERIC METHANE IN PRODUCTION SYSTEM:
LIFE CYCLE ASSESSMENT (LCA)
Enteric methane forms part of beef and dairy production
systems’ broader greenhouse gas (GHG) budget [60]. The
broader GHG budget includes methane, nitrous oxide (N2O),
and CO2 emissions from manure, feed production, vehicles and
transport, and other plants and equipment. The total GHG bud-
get of a production system is determined by life cycle assess-
ment (LCA) methodology, standardized by ISO 14040 [61] and
ISO 14044 [62] ([63, 64]).
Recently, studies were carried out to evaluate the impact
of bu󰀨alo production on greenhouse gas emissions [65, 66, 57,
67].
In collaboration with Embrapa Research Institute and
Cargill, our research group has studied the LCA of a bu󰀨alo
milk production farm in Brazil. The data showed that enteric
methane produced by bu󰀨aloes is the most relevant source of
GHG production. One estimate of LCA for bu󰀨alo milk produc-
tion was 63.4% for enteric methane (CH4), 33.9% for feed pro-
duction (CO2, N2O), and 1.92% for manure (FIG. 2).
As enteric methane has the most signicant impact on
the production of CO2 equivalent, the reduction in production
cycles (reduction in age at rst calving and the interval be-
tween calving) and an increase in individual production (milk
and meat) contribute signicantly to the farm’s sustainability.
Furthermore, new technologies can potentially manipulate the
rumen microbiome through genetic selection and various di-
etary intervention strategies to reduce CH4 emissions. Accord-
ing to Yusuf et al. [68], methane reduction strategies have been
grouped into three crucial factors: management, nutrition, and
the use of advanced biotechnology. Manipulation of the rumen
in reducing methane using chemicals, feed additives, rough-
age, concentrate utilization, and plants containing secondary
compound oils has been reported [5]. Using technologies to
reduce methane emissions from these crucial factors in the
production chain will considerably impact the sustainability of
bu󰀨alo farming).
Studies in Italy found that, despite the methane pro-
duction on bu󰀨alo farms, the amount of greenhouse gases
converted into CO2-eq emitted during the bu󰀨alo dairy pro-
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duction system is lower than the CO2-eq removed from the at-
mosphere [59]. The authors found that for every kg of bu󰀨alo
Mozzarella cheese produced, 52 kg of CO2-eq is subtracted
from the atmosphere (di󰀨erences between CO2-eq emissions
from the production system and CO2-eq removal from the at-
mosphere).
This information demonstrates that it is possible to pro-
duce beef and bu󰀨alo meat in balance with the environment,
if appropriate technologies based on scientic information are
used.
CONCLUSIONS AND FUTURE DIRECTION
The global attention on enteric CH4 production in bu󰀨a-
loes requires a response that involves collaboration between
researchers and industry. Future generations of bu󰀨aloes will
be characterized by better e󰀩ciency and fertility, which may re-
duce CH4 emission intensity. This will result from a balanced
multi-trait selection and improved management. Articial in-
semination can be incorporated into bu󰀨alo breeding programs
to further improve reproductive e󰀩ciency and genetic gain, col-
laborating to reduce CO2-eq emission intensity. The urgency in
moving to the next generation of bu󰀨aloes will increase the pro-
duction of embryos from genomically dened prepubertal heif-
ers. This will reduce generation interval and accelerate the rate
of genetic improvement to bu󰀨aloes dened by better e󰀩ciency
and fertility and lower CH4 emission. The growing importance
of bu󰀨aloes in the world requires that they undergo an accel-
erated rate of genetic gain for e󰀩ciency of production, product
quality, and sustainability. The challenge remains to develop
integrated sustainable meat and milk production systems and
communicate the importance of bu󰀨aloes for food security and
the environment.
REFERENCES
[1] United Nations Department of Economic and Social
A󰀨airs, Population Division. World Population Prospects
2022: Summary of Results. UN DESA/POP/2022/TR/
NO. 3.
[2] FAO. Sustainable Development Goals (SDG). End hun-
ger, achieve food security, and improved nutrition, and
promote sustainable agriculture. 2019. Available: http://
www.fao.org/sustainable-development-goals/news/de-
tail-news/en/c/424259/
[3] Zhang Y, Colli L, Barker JSF. Asian water bu󰀨alo: Domes-
tication, history and genetics. Animal Genetics. 2020;
51(2): 177-191. https://doi.org/10.1111/age.12911
[4] FAO. Crops and livestock products. License: CC BY-NC-
SA 3.0 IGO. Extracted from: https://www.fao.org/faostat/
en/#data/QCL. Data of Access: 27-09-2023.
FIGURE 2. Main emission pathways related to a bu󰀨alo dairy farm in Brazil, quantied by life cycle assessment (LCA) meth-
odology [Methodology standardized by ISO 14040 (ISO, 2006a) and ISO 14044 (ISO, 2006b); de Vries et al., 2015; Kyttä et al.,
2022]. On-farm emissions represent 83.2% (agriculture activities), and o󰀨-farm emissions represent 16.8% (food produc-
tion, fertilizer production, and fuel and electricity; Adapted from Abreu et al., 2023; data not yet published)
88
13th World Bu󰀨alo Congress ~ 13er Congreso Mundial de Búfalos / Lectures / Reproduction ______________________________________
[5] Wanapat M, Kang SC. World bu󰀨alo production: Challen-
ges in meat and milk production, and mitigation of metha-
ne emission. Bu󰀨alo Bulletin. 2013, 32(1): 1–21.
[6] Knapp Jr B, Nordskog AW. Heritability of growth and e󰀩-
ciency in beef cattle. Journal of Animal Science. 1946;
5(1): 62-70. https://doi.org/10.2527/jas1946.5162
[7] Faverdin P, Guyomard H, Puillet L, Forslund A. Animal
board invited review: Specialising and intensifying cattle
production for better e󰀩ciency and less global warming:
contrasting results for milk and meat co-production at di-
󰀨erent scales. Animal. 2022; 16(1): 100431. https://doi.
org/10.1016/j.animal.2021.100431
[8] Baruselli PS, Abreu LA, Paula VR, Carvalho B, Gricio EA,
Mori FK, Rebeis LM, Albertini S, Souza AH, D’Occhio MJ.
Applying assisted reproductive technology and reproduc-
tive management to reduce CO2-equivalent emission in
dairy and beef cattle: a review. Animal Reproduction.
2023; 20(2): e20230060. https://doi.org/10.1590/1984-
3143-AR2023-0060
[9] Galyean ML, Hales KE. Feeding management strategies
to mitigate methane and improve production e󰀩ciency
in feedlot cattle. Animals. 2023; 13(4): 758. https://doi.
org/10.3390/ani13040758
[10] Chang J, Peng S, Ciais P, Saunois M, Dangal SRS, He-
rrero M, Havlík P, Tian H, Bousquet P. Revisiting enteric
methane emissions from domestic ruminants and their
δ13CCH4 source signature. Nature Communications.
2019; 10: 3420. https://doi.org/10.1038/s41467-019-
11066-3
[11] FAO. Reducing enteric methane for improving food se-
curity and livelihoods. 2021. Available: http://www.fao.
org/in-action/enteric-methane/background/why-is-ente-
ric-methane-important/en/
[12] Bowen JM, Cormican P, Lister SJ, McCabe MS, Duthie
CA, Roehe R, Dewhurst RJ. Links between the rumen
microbiota, methane emissions and feed e󰀩ciency of
nishing steers o󰀨ered dietary lipid and nitrate supple-
mentation. PLoS One. 2020; 15(4): e0231759. https://
doi.org/10.1371/journal.pone.0231759
[13] Ross EM, Moate PJ, Marett L, Cocks BG, Hayes BJ. In-
vestigating the e󰀨ect of two methane-mitigating diets on
the rumen microbiome using massively parallel sequen-
cing. Journal of Dairy Science. 2013; 96(9): 6030-6046.
https://doi.org/10.3168/jds.2013-6766
[14] Knapp JR, Laur GL, Vadas PA, Weiss WP, Tricarico JM.
Invited review: Enteric methane in dairy cattle produc-
tion: Quantifying the opportunities and impact of reducing
emissions. Journal of Dairy Science. 2014; 97(6): 3231-
3261. https://doi.org/10.3168/jds.2013-7234
[15] Roehe R, Dewhurst RJ, Duthie CA, Rooke JA, McKain N,
Ross DW, Hyslop JJ, Waterhouse A, Freeman TC, Wat-
son M, Wallace RJ. Bovine host genetic variation inuen-
ces rumen microbial methane production with best selec-
tion criterion for low methane emitting and e󰀩ciently feed
converting hosts based on metagenomic gene abundan-
ce. PLoS Genetics. 2016; 12(2): e1005846. https://doi.
org/10.1371/journal.pgen.1005846
[16] Baruselli PS, Carvalho NAT, Gasparrini B, Campanile
G, D’Occhio MJ. Review: Development, adoption, and
impact of assisted reproduction in domestic bu󰀨aloes.
Animal. 2023; 17(S1): 100764. https://doi.org/10.1016/j.
animal.2023.100764
[17] Malik PK, Trivedi S, Mohapatra A, Kolte AP, Sejian V,
Bhatta R, Rahman H. Comparison of enteric methane
yield and diversity of ruminal methanogens in cattle and
bu󰀨aloes fed on the same diet. PLoS ONE. 2021; 16(8):
e0256048. https://doi.org/10.1371/journal.pone.0256048
[18] Baruselli, PS, Carvalho JGS, Elli󰀨 FM, Da Silva JCB,
Chello D, Carvalho NAT. Embryo transfer in bu󰀨alo (Bub-
alus bubalis). Theriogenology. 2020; 150: 221-228. ht-
tps://doi.org/10.1016/j.theriogenology.2020.01.037
[19] Hegarty RS, McEwan JC. Genetic opportunities to redu-
ce enteric methane emissions from ruminant livestock.
In ‘Proceedings of the Ninth World Congress on Ge-
netics Applied to Livestock Production’; 2010; Leipzig,
Germany. Leipzig: German Society for Animal Science.
2010. pp 181-186.
[20] Baruselli PS, Carvalho NAT, Gimenes LU, Crepaldi GA.
Fixed-time articial insemination in bu󰀨alo. Italian Jour-
nal of Animal Science. 2007; 6(S2): 107-118. https://doi.
org/10.4081/ijas.2007.s2.107
[21] Zicarelli L. Reproductive seasonality in bu󰀨alo. Bubalus
Bubalis. 1997; 4(29): 52–54.
[22] D’Occhio MJ, Ghuman SS, Neglia G, Della Valle G, Ba-
ruselli PS, Zicarelli L, Visintin JA, Sarkar M, Campanile
G. Exogenous and endogenous factors in seasonality of
reproduction in bu󰀨alo: A review. Theriogenology. 2020;
150: 186-192. https://doi.org/10.1016/j.theriogenolo-
gy.2020.01.044
[23] Zicarelli L. Can we consider bu󰀨alo a non precocious and
hypofertile species? Italian Journal of Animal Science.
2007; 6(S2): 143-154. https:
[24] Carvalho NAT, Soares JG, Porto Filho RM, Gimenes
LU, Souza DC, Nichi M, Sales JS, Baruselli PS. Equine
chorionic gonadotropin improves the e󰀩cacy of a timed
articial insemination protocol in bu󰀨alo during the non-
breeding season. Theriogenology. 2013; 79(3): 423-428.
https://doi.org/10.1016/j.theriogenology.2012.10.013
[25] Monteiro BM, Souza DC, Vasconcellos GSFM, Carval-
ho NAT, Baruselli PS. E󰀨ect of season on dairy bu󰀨alo
reproductive performance when using P4/E2/eCG-based
xed-time articial insemination management. Therio-
89
________________________________________________________ Revista Cientíca, FCV-LUZ / Vol. XXXIII, Supl. Esp., 82 - 91, 2023
genology. 2018; 119: 275-281. https://doi.org/10.1016/j.
theriogenology.2018.07.004
[26] Baruselli PS, Madureira EH, Barnabe VH, Barnabe RC,
Berber RCA. Evaluation of synchronization of ovulation
for xed timed insemination in bu󰀨alo (Bubalus buba-
lis). Brazilian Journal of Veterinary Reaserch and Animal
Science. 2003; 40(6): 431-442. https://doi.org/10.1590/
S1413-95962003000600007
[27] Sá Filho MF, Carvalho NAT, Gimenes LU, Torres-Júnior
JR, Nasser LFT, Tonhati H, Garcia JM, Gasparrini B, Zi-
carelli L, Baruselli, PS. E󰀨ect of recombinant bovine so-
matotropin (bST) on follicular population and on in vitro
bu󰀨alo embryo production. Animal Reproduction Scien-
ce. 2009; 113(1-4): 51-59. https://doi.org/10.1016/j.anire-
prosci.2008.06.008
[28] Carvalho JGS, Carvalho NAT, Bayeux BM, Watanabe YF,
Watanabe OY, Mingoti RD, Baruselli PS. Superstimula-
tion prior to the ovum pick-up improves the in vitro em-
bryo production in nulliparous, primiparous and multipa-
rous bu󰀨alo (Bubalus bubalis) donors. Theriogenology.
2019; 138: 164-168. https://doi.org/10.1016/j.theriogeno-
logy.2019.07.003
[29] Silva JCB, Rezende RG, Colli MHA, Bayeux, BM, Mingo-
ti RD, Ojeda-Rojas OA, Basso AC, Naves JR, Baruselli
PS. In vitro embryo production in bu󰀨alo: Comparison
between calves, prepubertal heifers and lactating cows.
Animal Reproduction. 2017; 14(3): 766.
[30] Baldassarre H, Bordignon V. Laparoscopic ovum pick-up
for in vitro embryo production from dairy bovine and bu-
󰀨alo calves. Animal Reproduction. 2018; 15(3): 191-196.
https://doi.org/10.21451/1984-3143-AR2018-0057
[31] Baruselli PS, Soares JG, Bayeux BM, Da Silva JCB,
Mingoti RD, Carvalho NAT. Assisted reproductive tech-
nologies (ART) in water bu󰀨aloes. Animal Reproduction.
2018; 15(1): 971-983. http://dx.doi.org/10.21451/1984-
3143-AR2018-0043
[32] Currin L, Baldassarre H, Bordignon V. In vitro production
of embryos from prepubertal Holstein cattle and Medi-
terranean water bu󰀨alo: Problems, progress and poten-
tial. Animals. 2021; 11(8): 2275. https://doi.org/10.3390/
ani11082275
[33] Baruselli PS, Rodrigues CA, Ferreira RM, Sales JNS,
Elli󰀨 FM, Silva LG, Viziack MP, Factor L, D’Occhio MJ.
Impact of oocyte donor age and breed on in vitro embryo
production in cattle, and relationship of dairy and beef
embryo recipients on pregnancy and the subsequent
performance of o󰀨spring: A review. Reproduction, Fer-
tility and Development. 2021; 34(2): 36-51. https://doi.
org/10.1071/RD21285
[34] Cammack KM, Austin KJ, Lamberson WR, Conant GC,
Cunningham HC. Ruminnat Nutrition Symposium: Tiny
but mighty: The role of the rumen microbes in livestock
production. Journal of Animal Science. 2018; 96(2): 752-
770. https://doi.org/10.1093/jas/skx053
[35] Johnson DE, Ward GM. Estimates of animal methane
emissions. Environmental Monitoring and Assessment.
1996; 42: 133-141. https://doi.org/10.1007/BF00394046
[36] Arthur PF, Herd RM. E󰀩ciency of feed utilisation by lives-
tock - Implications and benets of genetic improvement.
Canadian Journal of Animal Science. 2005; 85(3): 281-
290. https://doi.org/10.4141/A04-062.
[37] Berry DP, Crowley JJ. Cell Biology Symposium: Gene-
tics of feed e󰀩ciency in dairy and beef cattle. Journal
of Animal Science. 2013; 91(4): 1594-1613. https://doi.
org/10.2527/jas2012-5862
[38] Gonzalez-Recio O, Pryce JE, Haile-Mariam M, Hayes
BJ. Incorporating heifer feed e󰀩ciency in the Austra-
lian selection index using genomic selection. Journal
of Dairy Science. 2014; 97(6): 3883-3893. https://doi.
org/10.3168/jds.2013-7515
[39] Sypniewski M, Strabel T, Pszczola M. Genetic variabi-
lity of methane production and concentration measured
in the breath of Polish Holstein-Friesian cattle. Animals.
2021; 11(11): 3175. https://doi.org/10.3390/ani11113175
[40] Perera BMAO. Reproductive cycles of bu󰀨alo. Animal
Reproduction Science. 2011; 124(3-4): 194-199. https://
doi.org/10.1016/j.anireprosci.2010.08.022
[41] Khan FA, Das GK, Pande M, Sarkar M, Mahapatra RK,
Shankar U. Alterations in follicular uid estradiol, pro-
gesterone and insulin concentrations during ovarian
acyclicity in water bu󰀨alo (Bubalus bubalis). Animal Re-
production Science. 2012; 130(1-2): 27-32. https://doi.
org/10.1016/j.anireprosci.2011.12.020
[42] Abdoon AS, Gabler C, Holder C, Kandil OM, Einspanier
R. Seasonal variations in developmental competence
and relative abundance of gene transcripts in bu󰀨a-
lo (Bubalus bubalis) oocytes. Theriogenology. 2014;
82(8): 1055-1067. https://doi.org/10.1016/j.theriogenolo-
gy.2014.07.008
[43] Phogat JB, Pandey AK, Singh I. Seasonality in bu󰀨aloes
reproduction. International Journal of Plant and Animal
Sciences. 2016; 6(2): 46-54.
[44] Sabia E, Napolitano F, Claps S, Braghieri A, Piazzolla,
N, Pacelli C. Feeding, nutrition and sustainability in dairy
enterprises: the case of Mediterranean bu󰀨aloes (Bub-
alus bubalis). In: Vastola, A. (Ed.), The Sustainability of
Agro-food and Natural Resource Systems in the Medite-
rranean Basin. 2015. Springer Open, pp. 57–64. https://
doi.org/10.1007/978-3-319-16357-4_5
[45] Sabia E, Claps S, Napolitano F, Annicchiarico G, Bruno
A, Francaviglia R, Sepe L, Aleandri R. In vivo digestibility
90
13th World Bu󰀨alo Congress ~ 13er Congreso Mundial de Búfalos / Lectures / Reproduction ______________________________________
of two di󰀨erent forage species inoculated with arbuscular
mycorrhiza in Mediterranean red goats. Small Ruminant
Research. 2015; 123: 83-87. https://doi.org/10.1016/j.
smallrumres.2014.10.008
[46] Sabia E, Napolitano F, Salvatore C, De Rosa G, Bari-
le VL, Braghieri A, Pacelli C. Environmental impact of
dairy bu󰀨alo heifers kept on pasture or in connement.
Agricultural Systems. 2018; 159: 42-49. https://doi.or-
g/10.1016/j.agsy.2017.10.010
[47] Sabia E, Napolitano F, De Rosa G, Terzano GM, Barile
VL, Braghieri A, Pacelli C. E󰀩ciency to reach age of pu-
berty and behaviour of bu󰀨alo heifers (Bubalus bubalis)
kept on pasture or in connement. Animal. 2014; 8(11):
1907-1916. https://doi.org/10.1017/S1751731114001876
[48] Borghese A. Bu󰀨alo Production and Research. Technical
Series 67. 2005. Rome, Italy: Food and Agriculture Orga-
nization of the United Nations.
[49] Baruselli OS, Barnabe VH, Barnabe RC, Visintin JA, Mo-
lero-Filho JR, Porto R. E󰀨ect of body condition score at
calving on postpartum reproductive performance in bu-
󰀨alo. Bu󰀨alo Journal. 2001; 1: 53-65.
[50] Usmani RH, Dailey RA, Inskeep EK. E󰀨ects of limited
suckling and varying prepartum nutrition on postpartum
reproductive traits of milked bu󰀨aloes. Journal of Dairy
Science. 1990; 73(6): 1564-1570. https://doi.org/10.3168/
jds.S0022-0302(90)78826-1
[51] Nanda AS, Brar PS, Prabhakar S. Enhancing reproduc-
tive performance in dairy bu󰀨alo: Major constraints and
achievements. Reproduction Supplement. 2003; 61: 27-
36.
[52] Perera BMAO, de Silva LNA, Kuruwita VY, Karunarat-
ne AM. Postpartum ovarian activity, uterine involution
and fertility in indigenous bu󰀨aloes at a selected villa-
ge location in Sri Lanka. Animal Reproduction Science.
1987; 14(2): 115-127. https://doi.org/10.1016/0378-
4320(87)90091-1
[53] Mohan V, Kuruwita VY, Perera BMAO, Abeygunawarde-
na H. E󰀨ects of suckling on the resumption of post-par-
tum ovarian activity in bu󰀨aloes. Tropical Agricultural Re-
search. 1990; 2: 306-315.
[54] Singh AK, Brar PS, Nanda AS, Prakash BS. E󰀨ect of suc-
kling on basal and GnRH-induced LH release in post-par-
tum dairy bu󰀨aloes. Animal Reproduction Science. 2006;
95(3-4): 244-250. https://doi.org/10.1016/j.anirepros-
ci.2005.10.004
[55] Perera BMAO, Abeygunawardena H, Vale WG, Chan-
talakhana C. Bu󰀨alo. In: Livestock and Wealth Creation
– Improving the Husbandry of Animals Kept by Poor Peo-
ple in Developing Countries. Livestock Production Pro-
gramme. Natural Resources International Limited. UK.
2005; pp 451–471.
[56] Pirlo G, Caré S, Fantin V, Falconi F, Buttol P, Terzano
GM, Masoni P, Pacelli C. Factors a󰀨ecting life cycle as-
sessment of milk produced on 6 Mediterranean bu󰀨alo
farms. Journal of Dairy Science. 2014; 97(10): 6583-
6593. http://dx.doi.org/10.3168/jds.2014-8007
[57] O’Brien D, Shalloo L, Patton J, Buckley F, Grainger C, Wa-
llace M. A life cycle assessment of seasonal grass-based
and connement dairy farms. Agricultural Systems. 2012;
107: 33-46. https://doi.org/10.1016/j.agsy.2011.11.004
[58] Chirone R, Paulillo A, Salatino P, Salzano A, Cristofaro
B, Cristiano T, Campanile G, Neglia G. Life cycle as-
sessment of bu󰀨alo milk: A case study of three farms in
southern Italy. Journal of Cleaner Production. 2022; 365:
132816. https://doi.org/10.1016/j.jclepro.2022.132816
[59] Campanile G, Baruselli PS, Vecchio D, Prandi A, Neglia
G, Carvalho NAT, Sales JNS, Gasparrini B, D’Occhio MJ.
Growth, metabolic status and ovarian function in bu󰀨alo
(Bubalus bubalis) heifers fed a low energy or high energy
diet. Animal Reproduction Science. 2010; 22(1-2): 74-81.
https://doi.org/10.1016/j.anireprosci.2010.07.005
[60] De Vivo R, Zicarelli L, Napolano R, Zicarelli F. Calculation
method of the carbon footprint of products of animal ori-
gin integrated with the physiological absorption of carbon
dioxide: Calculation example of the CFP of mozzarella
di Bufala Campana DPO. Advances in Environmental
and Engineering Research. 2023; 4(3): 044. https://doi.
org/10.21926/aeer.2303044
[61] Ibidhi R, Calsamiglia S. Carbon footprint assessment
of Spanish dairy cattle farms: E󰀨ectiveness of dietary
and farm management practices as a mitigation strate-
gy. Animals. 2020; 10(11): 2083. https://doi.org/10.3390/
ani10112083
[62] ISO 14040. Environmental management - life cycle as-
sessment - principles and framework. 2006. Reference
number ISO 14040:2006(E).
[63] ISO 14044. Environmental management - life cycle as-
sessment - requirements and guidelines. 2006. Referen-
ce number ISO 14044:2006(E).
[64] de Vries M, van Middlelaar CE, de Boer IJM. Compa-
ring environmental impacts of beef production systems:
A review of life cycle assessments. Livestock Scien-
ce. 2015; 178: 279-288. https://doi.org/10.1016/j.livs-
ci.2015.06.020
[65] Kyttä V, Roitto M, Astaptsev A, Saarinen M, Tuomisto HL.
Review and expert survey of allocation methods used in
life cycle assessment of milk and beef. The International
Journal of Life Cycle Assessment. 2022; 27: 191-204. ht-
tps://doi.org/10.1007/s11367-021-02019-4
[66] Ijaz M, Goheer MA. Emission prole of Pakistan’s agri-
culture: past trends and future projections. Environment,
91
________________________________________________________ Revista Cientíca, FCV-LUZ / Vol. XXXIII, Supl. Esp., 82 - 91, 2023
Development and Sustainability. 2021; 23: 1668-1687.
https://doi.org/10.1007/s10668-020-00645-w
[67] Romano E, De Palo P, Tidona F, Maggiolino A, Braga-
glio A. Dairy bu󰀨alo Life Cycle Assessment (LCA) a󰀨ec-
ted by a management choice: The production of wheat
crop. Sustainability. 2021; 13(19): 11108. https://doi.
org/10.3390/su131911108
[68] Correddu F, Lunesu MF, Caratzu MF, Pulina G. Recal-
culating the global warming impact of italian livestock
methane emissions with new metrics. Italian Journal of
Animal Science. 2023; 22(1): 125-135. https://doi.or-
g/10.1080/1828051X.2023.2167616
[69] Yusuf RO, Noor ZZ, Abba AH, Abu Hassan MA, Mohd
Din MF. Greenhouse gas emissions: Quantifying metha-
ne emissions from livestock. American Journal of Engi-
neering and Applied Sciences. 2012; 5(1): 1-8. https://
doi.org/10.3844/ajeassp.2012.1.8