Original
Behavior, performance and carcass yield of steers with
different comfort conditioning during the fattening period
Comportamiento, desempeño y rendimiento de la canal
mediante la creación de condiciones de confort durante la ceba
María Eugenia Munilla *, Juan Sebastián Vittone
*, Andrea Biolatto *, Sonia Alejandra Romera **y***, Gustavo
Adolfo Teira ****
*Departamento de Rumiantes, Estación Experimental Agropecuaria, Instituto
Nacional de Tecnología Agropecuaria, Ruta Provincial 39 km 143.5, Concepción
del Uruguay (CP 3260), Entre Ríos, Argentina.
**Instituto de
Virología, Centro de Investigación en Ciencias Veterinarias y Agronómicas,
Instituto Nacional de Tecnología Agropecuaria, Nicolás Repetto y de los Reseros
s/n. (CP: 1686) Hurlingham, Buenos Aires, Argentina.
*** Consejo Nacional de Investigaciones
Científicas y Técnicas, Argentina.
**** Laboratorio de Industrias Cárnicas,
Universidad Nacional of Entre Ríos, Monseñor Tavella
1424 (CP: 3200), Concordia, Entre Ríos, Argentina.
Correspondencia:
munilla.maria@inta.gob.ar
Recibido: Octubre, 2023;
Aceptado: Noviembre, 2023; Publicado: Diciembre, 2023.
Objective. The aim of this
study was to evaluate the behavior and performance of steers in two different
housing areas (10 vs. 100 m2/animal) and two feeding models (daily
supply vs. self-feeding) during the fattening period. Materials and methods:
Forty-eight Hereford steers with initial live weight (LW): 214,0 kg were housed
under four treatments: 100DS (100 m2/animal and daily supply), 100SF
(100 m2/animal and self-feeding), 10DS (10 m2/animal and
daily supply), 10SF (10 m2/animal and self-feeding) until reaching a
final LW of 370 – 390 kg. Behavior was recorded by observation. Dry matter
intake (DMI) and average daily gain (ADG) were recorded to estimate feed
conversion ratio (FCR). Back fat thickness (BFT) and Longissimus muscle area (LMA) were measured. Steers were sent to a
commercial abattoir to evaluate carcass yield.
Results: The frequency of rest and walking was higher in steers
in the largest housing area. Self-feeding contributed to the intake
distribution during the day. No differences were detected in DMI, ADG and FCR.
However, the confined animals (10DS and 10SF) produced more BFT and less LMA.
In addition, they had less carcass yield at slaughter. Conclusions:
Therefore, a larger housing area and the self-feeding model could contribute to
express steers’ natural behavior and improve their performance.
KEYWORDS: animal welfare, cattle, feedlot,
housing area, feeding (Source: MESH)
Objetivo. EL propósito de
este estudio es evaluar el comportamiento y desempeño de bueyes en dos zonas de
estancia (10 vs. 100 m2/animales) y dos modelos de alimentación
(suplementación diaria vs autoalimentación) durante el periodo de ceba. Materiales
y métodos: Se emplearon un total de 48 bueyes Hereford con un
peso inicial (PI) de 214,0 kg con dos tratamientos diferentes: 100 SD (100 m2/animal
y suplementación diaria), 100 AA (100 m2/animal y auto
alimentación), 10 SD (10 m2/animal y suplementación diaria), 10 AA
(10 m2/animal y auto alimentación) hasta alcanzar un peso final (PF)
de 370 – 390 kg. Se registró el comportamiento mediante observación.
Igualmente, se registró el consumo de materia seca (CMS) junto a la ganancia
promedio diaria (GPD) para estimar la relación de conversión alimentaria (RCA).
Se midieron el grosor de la grasa posterior (GGP) y la zona muscular Longissimus (ZML) y se enviaron los bueyes al
matadero para evaluar el rendimiento de la canal. Resultados: La
frecuencia de descanso/marcha fue mayor en los bueyes que permanecieron en la
mayor área de estancia. La auto alimentación contribuyó a distribuir el consumo
de alimentos durante el día. No se detectaron diferencias en CMS, GPD y RCA.
Sin embargo, los animales confinados (10 SD y 10 AA) produjeron más GGP y menos
de ZML. Por otra parte, mostraron un menor rendimiento de la canal durante el
sacrificio. Conclusiones: Un área de estancia mayor, junto al modelo de
autoalimentación, contribuyó a un comportamiento natural de los bueyes y un
mejoramiento de su desempeño.
Palabras claves: bienestar animal,
ganado bovino, nave, área de estancia, alimentación (Fuente: MESH)
Beef cattle
is mostly fattened under intensive production systems that are generally
restrictive in terms of access to valuable resources such as living space,
freedom of movement and interaction with natural substrates. In many countries,
they are housed in places where muddy condition is a major problem for animal
welfare (Grandin, 2016).
Despite the
fact that confinement increases emerging diseases and transmission of
pre-existing diseases (Rossanigo et al., 2009), it is a widely used alternative for cattle
fattening. However, due to the multifactorial influence, confinement may not
achieve higher yields compared to grasslands or semi-confinement systems. In
addition, it is important to highlight that consumers
increasingly demand better quality, good production practices, animal welfare,
traceability and sustainability (Mota and Marçal, 2019).
The first
method to keep cattle clean is a correct stocking density (Grandin, 2016). Mader and Colgan (2007) found that lower cattle density in
feedlots resulted in lower muddy conditions. These results indicate that more
space could improve animal comfort and performance.
Space
influences feedlot cattle behavior and there is evidence that housing
modifications could promote livestock welfare (Park et al., 2020). Livestock behavior and welfare play an important
role in body development and carcass composition of beef cattle. Physical
comfort and nutritional conditions integrate the five domains model: nutrition,
environment, health, behavior, and mental state (Mellor et al., 2020).
Moreover,
delivering feed daily is not an option for producers with limited time or
equipment. Feed availability is a major limiting factor for production and
animal welfare. Feeding characteristics associated with low ruminal fluid pH
are: high dry matter intake and ingestion of large meals. It is because of the
greater amount of acid production per period of time, high eating rate because
of lower feed insalivation, short time spent chewing while eating and
ruminating because of lower daily saliva production, and large variations in
feeding behavior patterns throughout the day such as less frequent meals and
rumination. Adaptation of feeding behavior to diets with greater proportion of
concentrates also plays an important role, as smaller meals and more even
distribution of intake throughout the day lead to a better synchronization in
time between acid production and elimination or neutralization (González et al., 2012). Many farmers feed their
livestock only once a day to minimize the cost of labor. Self-feeders can be
used to provide ad libitum food. Final weight and yield are indicators of
animal welfare (Park et al., 2020).
Because of high concentrate diets and sedentary lifestyles cattle in
confinement could be prone to display poor health (Macitelli
et al., 2020). Feeding behavior can
also improve performance. An increase in feeding frequency during the fattening
period may contribute to promoting a better rumen environment for fermentation.
Furthermore, more stable ruminal conditions can decrease dry matter intake
(DMI) as feeding frequency increases (De Souza Teixeira et al., 2018). Therefore, the aforementioned feeding model and
homogeneous distribution of consumption along the day may contribute to express
innate behavior of displacement and rest in cattle (Oberschätzl
et al., 2016).
Farmers must
provide adequate conditions in order to meet livestock physiological and
behavioral needs (Fernandez-Novo et al., 2020). It is necessary to
promote a more sustainable and efficient production system than the current
agricultural and livestock systems. Thus, livestock conditions have to improve
animal welfare and reduce environmental problems. Based on this information,
the aim of this study was to evaluate behavior and performance in steers with
two housing areas (10 vs. 100 m2/animal) and two feeding models
(daily supply vs. self-feeding) during the fattening period.
MATERIALS AND METHODS
This research
was carried out at the INTA Agricultural Experimental Station, located in the
city of Concepción del Uruguay, Entre Ríos, Argentina (32°48’S, 58°34W).
Forty-eight Hereford steers with an initial age of 9 months and live weight
(LW) of 214,0 ± 23,7 kg were housed using four treatments: 100DS (100 m2/animal
and daily supply), 100SF (100 m2/animal and self-feeding), 10DS (10
m2/animal and daily supply) and 10SF (10 m2/animal and
self-feeding).
Previous to
the experience, animals underwent 35-day adaptation period. The fattening
period diet was formulated with 77% whole corn, 20% ground corn, 2% slow
released urea (43% N) and 1% mineral supplement. Animals were fed according to
the assigned treatment. The daily supply treatments meant that 3.2% LW animals
were offered feed every day whereas animals’ feeders were filled every four
days in self-feeding treatments.
Steers were
identified with numbers written on both sides of their bodies. Behavioral data
was collected by observation performed during the first 7 days of the
experience. Animals were observed for one hour three times during the day
(09:00= morning, 13:00= noon and 17:00=afternoon). Food intake (access to
feeder) and displacement (resting, walking, static standing) were recorded by
registering the number of times the animals executed these activities.
LW was
registered at 14-day intervals in order to estimate average daily gain (ADG).
Intake was determined as the difference between the amount of food provided and
residual food, divided by the number of animals in each treatment. Feed
conversion (FC) was determined by the ratio intake/ADG. DMI and ADG were recorded
individually to determine the feed conversion ratio (FCR).
Back fat
thickness (BFT) and Longissimus
muscle area (LMA) were measured in steers at 28-day intervals until slaughter.
A real time ultrasound machine (FALCOVET 100, PieMedical,
Holland) was used to measure LMA and BFT between the 12th and 13th rib and
vegetable oil was used as a coupling agent.
Steers were
fattened to 370 – 390 kg LW and sent to a commercial abattoir. The hot carcass
weight was obtained after pelvic fat removal. Carcass yield was calculated by
dividing hot carcass weight by final body weight (7% dressing).
Statistical
analysis was performed using PROC GLM (4). The model includes the effect of two
housing areas (HA), two feeding (F) models and the interactions between these
effects (HA*F). The model also included the effect of time on data collection.
The steers were considered as experimental units (n= 12). When interaction or
principal factors were significant (p<0.05),
means were compared by the Tukey test.
RESULTS
AND DISCUSSION
A difference
in frequency of food intake was found among different feeding models along the
day (p<0.0001, Figure 1). The
animals with daily supply (100DS and 10DS) presented higher intake frequency in
the morning when compared to noon and afternoon (p<0.05). These treatments also showed higher intake frequency
regarding 100SF and 10SF in the morning, similar at noon and lower in the
afternoon. Self-feeding resulted in a homogeneous feeding pattern along the day
(p>0.05). The higher feeding
activity in the morning in 100DS and 10DS was associated with the conditioned
reflex caused by the food supply.
Figure 1.
Food intake along the day in steers with different comfort conditions during
the fattening period. 100DS: 100 m2/animal and daily supply, 100SF:
100 m2/animal and self-feeding, 10DS: 10 m2/animal and
daily supply, 10SF: 10 m2/animal and self-feeding.
a, b: Mean values represented by different letters in the rows indicate
statistical differences detected by the Tukey test (p<0.05).
Comfort conditions
(housing area and feeding model) affected displacement in steers during the
fattening period (Figure 2). Treatments with larger housing areas (100DS and
100SF) presented a higher walking frequency in the animals. Only 1% of confined
animals (10DS and 10SF) walked and also had a lower frequency of rest due to
the smaller space (p<0.05).
Figure. 2.
Displacement in steers with different comfort conditions during the fattening
period. 100DS: 100 m2/animal and daily supply, 100SF: 100 m2/animal
and self-feeding, 10DS: 10 m2/animal and daily supply, 10SF: 10 m2/animal
and self-feeding.
a, b: Mean values represented by different letters in the rows indicate
statistical differences detected by the Tukey test (p<0.05).
Patterns of
food intake agrees with the findings of Mattachini et al. (2011) who found that the
frequency of feed delivery affected the pattern of lying down? behavior
throughout the day and the lying down time following the provision of feed.
Under natural
conditions, cattle display a need of spreading out their feeding behavior over
the whole day (Schneider et al.,
2019). Results found in this work show a better intake behavior along the day
in animals on a self-feeding system. Displacement behavior results are similar
to Schütz et al.
(2019) who found that cows on manure contaminated surfaces had a reduced lying
time in comparison with those on dry soil. They suggested that the reduction in
lying down time is
predominantly due to the surface moisture content. Muddy surfaces have negative
effects on the health and welfare of dairy cattle, and if possible, cows will
avoid this surface.
Final weight,
ADG, TWG, DMI and FCR were similar regardless of comfort conditions during the
fattening period (Table 1).
Table 1.
Mean values for weight evolution, dry matter intake and feed conversion ratio
in steers with different comfort conditions during the fattening period.
|
100DS |
100SF |
10DS |
10SF |
SEM |
p |
||
HA |
F |
HA*F |
||||||
IW1
(kg) |
214.9 |
212.8 |
220.0 |
208.3 |
3.42 |
0.97 |
0.32 |
0.49 |
FW2
(kg) |
369.9 |
371.8 |
384.2 |
376.8 |
2.98 |
0.11 |
0.65 |
0.44 |
ADG3 (kg) |
1.3 |
1.3 |
1.3 |
1.4 |
0.02 |
0.13 |
0.33 |
0.97 |
TWG4 (kg) |
155.0 |
159.1 |
164.2 |
168.5 |
3.91 |
0.41 |
0.70 |
0.99 |
DMI5 (kg) |
1103.8 |
1095.3 |
1106.4 |
1139.4 |
25.43 |
0.41 |
0.70 |
0.99 |
FCR6 (kg) |
7.2 |
6.9 |
6.8 |
6.8 |
0.16 |
0.58 |
> 0.99 |
0.73 |
100DS: 100 m2/animal and daily supply,
100SF: 100 m2/animal and self-feeding, 10DS: 10 m2/animal
and daily supply, 10SF: 10 m2/animal and self-feeding, HA: housing
area, F: feeding models.
1Initial weight, 2Final weight, 3Average daily
gain, 4Total Weight gain, 5Dry matter intake, 6Feed
conversion ratio.
Ultrasound
measurement and carcass yield were affected by comfort conditions (Table 2). A
significant difference in final and total gain BFT (p<0.05) according to housing areas was observed. At the end of
the fattening period, the treatment 10DS showed a higher BFT than 100SF.
Conversely, LMA final and total gain was higher for 100SF when compared to
10SD. Housing areas also affected carcass yield. Animals with more housing
space presented higher values (p=0.0010).
Table 2.
Mean values for ultrasound measurement and carcass yield in steers with
different comfort conditions during the fattening period.
|
100DS |
100SF |
10DS |
10SF |
SEM |
P |
||
HA |
F |
HA*F |
||||||
Initial BFT1
(mm) |
6.3 |
6.2 |
6.4 |
6.4 |
0.12 |
0.46 |
0.71 |
0.76 |
Final BFT (mm) |
10.7 ab |
10.4 a |
11.6 b |
11.0 ab |
0.18 |
0.03 |
0.21 |
0.73 |
Total gain BFT
(mm) |
4.4 ab |
4.2 a |
5.2 b |
4.6 ab |
0.16 |
0.04 |
0.29 |
0.54 |
Initial LMA2
(cm2) |
40.3 |
39.6 |
43.3 |
40.1 |
0.98 |
0.38 |
0.34 |
0.53 |
Final LMA (cm2) |
76.8 ab |
80.6 a |
74.2 b |
75.0 ab |
1.12 |
0.004 |
0.29 |
0.48 |
Total gain LMA
(cm2) |
36.5 ab |
41.0 a |
30.9 b |
34.8 ab |
1.50 |
0.03 |
0.15 |
0.93 |
Hot carcass
weight (kg) |
212.5 |
213.4 |
214.5 |
211.7 |
1.87 |
0.72 |
0.57 |
0.42 |
Carcass yield (%) |
61.7 a |
61.7 a |
60.1 b |
60.4 b |
0.00 |
<0.001 |
0.93 |
0.92 |
1Back fat thickness, 2Longissimus
muscle area.
100DS: 100 m2/animal and daily supply, 100SF: 100 m2/animal
and self-feeding, 10DS: 10 m2/animal and daily supply, 10SF: 10 m2/animal
and self-feeding.
a, b: Mean values represented by different letters in the rows indicate
statistical differences detected by the Tukey test (p<0.05).
According to
Park et al. (2020), space allowance
influences cattle behavior and performance and they are indicators of positive
welfare state. Exposure to mud also has implications for cattle hygiene and
health. In this research, a smaller housing area produced muddy conditions
which resulted in poor animal hygiene (Photo 1). With more space, animals
remained clean during all the fattening period (Photo 2). Prior rainfall and
surface water pooling were useful measures to determine less lying down time,
and thus animal welfare, are compromised (Neave et al., 2022).
Chen et al. (2015) suggest that poor hygiene
could present an increased risk of infection and immunosuppression. In
addition, Macitelli et al. (2020) found decreasing the space allowance for beef cattle
in outdoor feedlots degrades the feedlot environment and affect animal welfare.
Photo 1 and 2. Steers with different housing areas (left: 10 m2/animal,
right: 100 m2/animal) during the fattening period.
Accordingly, Pordomingo et al.
(2022) reported similar live weight evolution and feed efficiency in feedlot
cattle. However, Grandin (2022) and Mader and Griffin
(2015) found efficiency problems with confinement and muddy conditions.
According to
De Souza Teixeira et al. (2018),
intake, ADG and FCR were not impacted by behavior (food intake along the day
and displacement). However, in this work, behavior affected fat deposition, LMA
growth and carcass yield. It was found a high correlation between LMA and
carcass yield (p<0.05). Carcass yield presented a significant positive
correlation (p<0.05) with walking (r= 0.44) and resting (0.43). A positive
canonical correlation (p=0.03) was also determined between behavior associated
with animal welfare (walking and resting) and carcass yield. On the other hand,
Dunston-Clarke et al. (2020) showed
that sedentary cattle had lower carcass yield. In this work, similar results
about behavior and its effects on fattening and carcass yield were observed.
CONCLUSIONS
More space allowance on the feedlot pens provides
a better environment for the animals, offering them more choices on where to
stay or lay down, and reducing the risk of diseases spread. Animals with more
space walk and rest for longer periods. Improved comfort associated to
self-feeding contributes to deployment of natural behavior of cattle during the
fattening period. Self-feeding contributes to meal frequency along the day
without affecting the animal performance.
Nevertheless, confinement increases fattening and
reduces the Longissimus muscle area,
with lower carcass yield. Larger housing areas and self-feeding under the
Ecological Feedlot system are an alternative to intensive fattening systems.
This study is valuable for farmers to improve animal welfare and carcass yield
in cattle. Further exploration to develop a suitable system that is repeatable
is recommended.
ACKNOWLEDGMENTS
To the
Argentine Beef Promotion Institute (IPCVA) for financial support.
REFERENCIAS
Chen, Y., Arsenault, R., Napper, S., & Griebel, P. (2015). Models and methods to investigate acute
stress responses in cattle. Animals, 5(4), 1268-1295. https://www.mdpi.com/2076-2615/5/4/411
De Souza Teixeira, O., Brondani,
I. L., Alves Filho, D. C., Nörnberg, J. L., Cattelam, J., Pereira, L. B., & Klein, J. L. (2018).
Performance and ingestive and social behavior of
young cattle with different sexual conditions supplemented in Aruana pasture. Semina:
Ciências Agrárias, 39(6),
2565-2580. https://ojs.uel.br/revistas/uel/index.php/semagrarias/article/view/31105
Dunston-Clarke, E. J., Hunter, I., & Collins, T.
(2020). Influence of Exercise Enrichment on Feedlot Cattle Behaviour
and the Human–Animal Relationship. Proceedings, 73 (4), 2-7. https://www.mdpi.com/2504-3900/73/1/4
Fernandez-Novo,
A., Pérez-Garnelo, S. S., Villagrá, A.,
Pérez-Villalobos, N., & Astiz, S. (2020). The effect of stress on reproduction and reproductive
technologies in beef cattle—A review. Animals, 10(11),
2096. https://www.mdpi.com/2076-2615/10/11/2096
González, L. A., Manteca, X., Calsamiglia,
S., Schwartzkopf-Genswein, K. S., & Ferret, A.
(2012). Ruminal acidosis in feedlot cattle: Interplay between feed ingredients,
rumen function and feeding behavior (a review). Animal feed science and
technology, 172(1-2), 66-79. https://www.sciencedirect.com/science/article/abs/pii/S0377840111004986
Grandin, T. (2016). Evaluation of the welfare of
cattle housed in outdoor feedlot pens. Veterinary and Animal Science, 1,
23-28. https://www.sciencedirect.com/science/article/pii/S2451943X16300278
Grandin, T. (2022). Practical Application of the Five
Domains Animal Welfare Framework for Supply Food Animal Chain Managers. Animals, 12(20),
2831. https://www.mdpi.com/2076-2615/12/20/2831
Macitelli,
F., Braga, J. S., Gellatly, D., & da Costa, M. P.
(2020). Reduced space in
outdoor feedlot impacts beef cattle welfare. animal, 14(12),
2588-2597. https://www.cambridge.org/core/journals/animal/article/abs/reduced-space-in-outdoor-feedlot-impacts-beef-cattle-welfare/C94CC107B54CF37A185357B23BE9B1FD
Mader, T. L., & Griffin, D. (2015). Management of
cattle exposed to adverse environmental conditions. Veterinary Clinics:
Food Animal Practice, 31(2), 247-258. https://www.vetfood.theclinics.com/article/S0749-0720(15)00021-3/fulltext
Mader, T. L., & Colgan, S. L. (2007). Pen density and
straw bedding during feedlot finishing. Nebraska Beef
Cattle Reports, 70, 43-46. https://digitalcommons.unl.edu/animalscinbcr/70/
Mattachini, G., Riva, E., Pompe, J. C.
A. M., Bisaglia, C., & Provolo,
G. (2011). Methods for measuring the behaviour of
dairy cows in free stall barns. https://library.wur.nl/WebQuery/wurpubs/fulltext/195362
Mellor, D. J., Beausoleil,
N. J., Littlewood, K. E., McLean, A. N., McGreevy, P. D., Jones, B., &
Wilkins, C. (2020). The 2020 five domains model: Including human–animal
interactions in assessments of animal welfare. Animals, 10(10),
1870. https://www.mdpi.com/2076-2615/10/10/1870
Mota,
R. G., & Marcal, W. S. (2019). Comportamento e bem-estar animal
de bovinos confinados: Alternativas para uma produção eficiente, rentável e de
qualidade: Revisão
bibliográfica. Revista Brasileira de Higiene e Sanidade
Animal: RBHSA, 13(1), 125-141. https://dialnet.unirioja.es/servlet/articulo?codigo=6997432
Neave, H. W., Schütz, K. E.,
& Dalley, D. E. (2022). Behavior of dairy cows
managed outdoors in winter: Effects of weather and paddock soil
conditions. Journal of Dairy Science, 105(10),
8298-8315. https://www.sciencedirect.com/science/article/pii/S0022030222004404
Oberschätzl-Kopp, R., Haidn, B., Peis, R., Reiter, K., & Bernhardt, H. (2016, June).
Effects of an automatic feeding system with dynamic feed delivery times on the behaviour of dairy cows. In Proc. of CIGR-AgEng 2016 Conference, Aarhus, Denmark (pp. 1-8). https://www.cabdirect.org/cabdirect/abstract/20183376882
Park, R. M., Foster, M., & Daigle, C. L. (2020). A
scoping review: The impact of housing systems and environmental features on
beef cattle welfare. Animals, 10(4),
565. https://www.mdpi.com/2076-2615/10/4/565
Pordomingo,
A. J., Gelid, L., Pordomingo,
A. B., Baliño, P., & Bressan,
E. (2022). Uso de monensina y virginiamicina
en el engorde a corral de vaquillonas basado en maíz entero. RIA.
Revista de investigaciones agropecuarias, 48(1), 71-77. http://www.scielo.org.ar/scielo.php?pid=S1669-23142022000100071&script=sci_arttext
Rossanigo,
C. E., Bengolea, A., & Sager, R. L. (2009). Enfermedades bovinas en los sistemas
intensivos de la región semiárida-subhúmeda central. Revista Argentina de
Producción Animal, 29(2), 151-180. https://www.researchgate.net/publication/264544414_Enfermedades_bovinas_en_los_sistemas_intensivos_de_la_region_semiarida-subhumeda_central
Schneider, L., Kemper, N., & Spindler, B. (2019).
Stereotypic behavior in fattening bulls. Animals, 10(1),
40. https://www.mdpi.com/2076-2615/10/1/40
Schütz, K. E., Cave, V. M., Cox, N. R., Huddart, F. J.,
& Tucker, C. B. (2019). Effects of 3 surface types on dairy cattle
behavior, preference, and hygiene. Journal of dairy science, 102(2),
1530-1541. https://www.sciencedirect.com/science/article/pii/S0022030218311159
Research conception and design: MEM, JSV, AB, SAR, GAT;
data analysis and interpretation: MEM, JSV, AB, SAR, GAT; redaction of
the manuscript: MEM,
JSV, AB, SAR, GAT.
The authors state there are no conflicts of
interest whatsoever.