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Crudeli,G.A. Artificial Insemination in Buffalo in Northeastern Argentina. Pág. 1 © Copyright 2003. INTA. EEA Colonia Benítez. Marcos Briolini s/n (3505) Colonia Benítez, Chaco Argentina

Buffalo J. (1999) 1: 61-67 Original Article

Artificial Insemination in Buffalo in Northeastern Argentina

GA Crudeli, R.C. Stahringer*, P. Maldonado Vargas and M.S. Flores Barbaran

G.I.P.R.A. (Grupo de Investigación en Producción y Reproducción Animal), Facultad de Ciencias Veterinarias, U.N.N.E., Sargento Cabral 2139,( 3400) Corrientes, *EEA INTA Colonia Benítez, Chaco, Argentina

Received August 22, 1997

Accepted December 3, 1998

Key words:artificial insemination, heat sign, pregnancy rate, Argentina

Abstract

Thirty-six pluriparous buffalo cows with postpartum periods longer than 60 days were utilized. Cows were heat checked for two 90 minute periods in the motning and evening with the aid of 2 caudoepidectomized marker bulls. One bull was provided with a chinball marker and remained with the cows on pasture, while the other was only introduced to the cows during the heat observation periods in the holding pen. Buffalo cows were inseminated at first rejection to be mounted. All cows were diagnosed for pregnancy 60 days after the last insemination by rectal palpation. Eighty-six percent of the. buffalo cows was detected in estrus. The non return rate was 74.2%. Three percent of the cows had intervals from heat initiation to insemination shorter than 12 h, 87.9% between 12 and 24 h and 9.1% between 24 and 36 h. Mean heat duration was 18.8 h. Mean body condition score during the experiment was higher (3.5) in females detected in heat than in those in anestrus (2.5). Pregnancy rate was 74.2%. Difficulty score to pass the insemination gun through the cervix affected pregnancy rate. Cows c1assified with a high, medium and low difficulty score had pregnancy rates of 0, 80 and 73.1%, respectively.

Introduction

Buffaloes were introduced to Argentina fr9m the island of Marajo (Brazil) almost a century ago, but their numbers have not increased rapidly, accounting in the present to around 15,000 animals in our country, 60% of which are in the province of Corrientes. The herds of buffalo received little attention and some were used to provide specimens for hunting reserves. Difficulties were present to find

Buffalo Journal 15 (1)

Commercial channels for their carcasses. A lack of knowledge about their nutritional and reproductive physiology occurred. All these facts produced little interest in producers to attain genetic progress in their buffalo herds. For these reasons, buffalo herds show a high level of inbreeding determining smaller body weight, reduced fertility with observations of low scrotal circumference and small ovaries in the province of Corrientes (Crudeli, 1995; Crudeli, 1996 b,c). Artificial insemination has

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Crudeli,G.A. Artificial Insemination in Buffalo in Northeastern Argentina. Pág. 2 © Copyright 2003. INTA. EEA Colonia Benítez. Marcos Briolini s/n (3505) Colonia Benítez, Chaco Argentina

been a useful tool to achieve genetic improvement around the world. This is the second experience of our group using Al in buffalo. The objective was to obtain more information on reproductive behavior of buffaloes in order to develop practical management schemes to apply AI on the field.

Materials and Methods

The present work was carried out between May 16 and June 20, 1997 at the ranch "Rincon del Madregon", located 70 km south of the city of Corrientes in Empedrado County, 8 km East of Highway 12. During the experimental period, the animals were kept in a 70 ha paddock with natural pasture. Fifty percent of the paddock was permanently cover with water.

Animals :

Thirty-six pluriparous buffalo cows with postpartum periods longer than 60 days were selected. Most of them belonged to a herd imported from Brazil in 1990. Selection criteria were: previous fertility, body conformation and body condition score (based on the method developed for the bovine by D.B. Herd. and L.R. Sprott with 1 = emaciated and 9= obese and adapted for bubalines). Reproductive soundness of the females was given priority over conformation characteristics. All females and their calves were identified with numbered ear tags and additionally, buffalo cows had their number painted on the back. All females were serologically t~sted for brucellosis and were negative (B.P.A. and 2 mercapto-ethanol tests).

Marker bulls:

Two marker bulls (A: 2 years old; B: 3 years old) were used. They had been previously sterilized by a bilateral caudoepidectomy. Bull A was fitted with a chin ball marker and remained with the cow herd all day around. The bull B was kept on a separated paddock and introduced to the cow herd only during the heat detection periods at which time bull A was withdrawn from the cow herd.

Heat detection :

Heat was detected twice daily for 90 minutes periods from 7:00 to 8:30 AM and from 6:00 to 7:30 PM. The animals were gathered in a holding pen shortly before the afternoon heat-detection period and afterwards released to an adjacent small paddock to spend the night. They returned to the holding pen for the morning heat detection period and thereafter they were released to the pasture. Behavioral and external heat signs were taken into account for heat detection.

Artificial Insemination in Argentina Time of insemination:

Disappearance of heat signs (first rejection to be mounted by a marker bull) was established as the time of insemination. Females that showed up painted to any of the heat detection periods but rejected the mount of the marker bull were considered to

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Crudeli,G.A. Artificial Insemination in Buffalo in Northeastern Argentina. Pág. 3 © Copyright 2003. INTA. EEA Colonia Benítez. Marcos Briolini s/n (3505) Colonia Benítez, Chaco Argentina

have undergone heat during the period between observations (heat periods shorter than 12 h) and were also inseminated at that time.

Cows were inseminated with semen of Mediterranean breed bulls of proven fertility. Semen had been frozen in 0.25 mI Minitube straws containing 60 million sperms with a post-thawing motility of 60%. Straws were thawed in 40.C water for 30 seconds and semen was deposited into the body of the uterus.

Pregnancy diagnosis:

All cows were diagnosed for pregnancy 60 days after the last insemination by rectal palpation.

Statistical analysis:

Body condition scores were analysed as a complete randomized design utilizing the GLM procedure of SAS with estrus appearance as the main effect.

Results

A high percentage (86.1%, n = 31) of the buffalo cows was detected in heat. Twenty-three of these females did not return to heat after the first insemination (nonreturn rate to first insemination = 74.2%). Two females were reinseminated after returning to heat, but did not get pregnant to the second insemination.

Interval from heat initiation to insemination was shorter than 12 h in 1 cow (3.0%), between 12 and 24 h in 29 cows (87.9%) and between 24 and 36 h in 5 cows (9.1 %). Mean heat duration was 18.8 ± 3.3 h (mean ± standard deviation). The females which were not detected in heat by the marker bulls (n= 5; 13.9%) presented a prolonged postpartum anestrus. Three of them showed ovarian activity and 2 were diagnosed to have ovulated by rectal palpation. Mean body condition score during the heat detection period was higher (3.5 ± 0.4; P < 0.05) in females detected in heat than in those in anestrus (2.5 ± 0.3).

Pregnancy rate was 74.2% resulting in a ratio of 1.43 straws per pregnancy. Body condition score was similar in cows pregnant after insemination (3.5 ± 0.4) and non pregnant cows (3.4 ± 0.4).

The degree of difficulty to pass the insemination gun through the cervix affected pregnancy rates. Two cows were classified as high difficulty (6.1%), 5 as medium difficulty (15.1%) and 26 as low difficulty (78.8%). Pregnancy rates obtained in each group were 0%, 80% and 73.1%, respectively.

Discussion

Caudoepidectomized marker bulls had a better performance than previously observed by us in marker bulls provided with an apron hanging in front of the

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Crudeli,G.A. Artificial Insemination in Buffalo in Northeastern Argentina. Pág. 4 © Copyright 2003. INTA. EEA Colonia Benítez. Marcos Briolini s/n (3505) Colonia Benítez, Chaco Argentina

Prepucial orifice (Crudeli et al., ,1996). Caudoepidectomy has the inconvenience of allowing a normal service of the female by the marker bull, which can be avoided when marker bulls are submitted to lateral deviation of the penis (Vale, 1994a). But it showed to be an effective sterilization method of low cost and risk for the bull.

Interval from heat initiation to insemination in the present experiment (18.8 h) was similar to that observed by us previously (17.6 h; Crudeli et al., 1996). Vale (1988) observed a longer interval from heat initiation to insemination (21.7 h). Other authors have described shorter intervals from heat initiation to insemination of 13.1 h (Jacomini, 1989) or 14.7 h (Baruselli, 1992).

Pregnancy rates to first insemination were higher in the present work than those cited by other authors. In Argentina, Zava (1992) observed overall pregnancy rates of 38.8% in a herd of 18 buffalo cows and 62.5% in another herd of 8 females. In an earlier experiment by our group, we obtained a pregnancy rate of 28.5% in a herd that included virgin and adult females (Crudeli et al., 1996). Barnabe (1994) obtained pregnancy rates to first insemination of 56.8% utilizing semen frozen with the same protocol than the semen used by us. Baruselli (1994) observed a pregnancy rate to first insemination of 48.3% and an overall pregnancy rate of 71.8% after 2 inseminations with a use of 1.88 semen doses per pregnancy. In 3 different groups of buffalo females, Ribeiro et al. (1994) obtained pregnancy rates of 55.5%, 50% and 38.1 % with a calving rate of 67.1 % after 3 inseminations and an investment of 2.58 semen doses per pregnancy. Furthermore, Barnabe et al.

(1995) achieved 52.7% pregnancy rate with 1.9 semen doses per pregnancy.

Hegazy et al. (1994) found that body condition significantly influenced postpartum interval in buffalo cows. They observed that females in thin body condition presented a mean postpartum interval of 80.9 days while in females with moderate body condition, the duration was 59.3 days. But similarly to our results, these authors did not observe an effect of body condition on services per conception (thin = 1.34 vs moderate = 1.49). On the other side, Baruselli et al.

(1995) determined that body condition at parturition influenced postpartum interval, percentage of females in anestrus during the breeding season, number of inseminations per conception, breeding period and percentage of females pregnant at the end of the breeding season.

In our previous experiments, we observed a negative correlation between the difficulty score to pass the insemination gun through the cervix and percentage of females that became pregnant as observed in this experiment.

In Table 1, we present a summary of heat signs as detected by different authors in order to compare them with our heat detection experience in present and previous experiments.

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Crudeli,G.A. Artificial Insemination in Buffalo in Northeastern Argentina. Pág. 5 © Copyright 2003. INTA. EEA Colonia Benítez. Marcos Briolini s/n (3505) Colonia Benítez, Chaco Argentina

As result of the present experiment, we concluded that:

- We were able to develop an efficient heat detection system which is feasible to be carried out in the farm with an adequate identification of the insemination time allowing a high conception rate and a low number of semen doses per pregnancy.

- An 87.9% of the heat periods lasted between 12 to 24 h.

- Body condition at. the beginning of the breeding period affected estrus presentation but did not affect conception rate.

- We were able to show that artificial insemination achieved good fertility rates in buffalo cows where the degree of difficulty to pass the cervix was low to moderate.

References

Baruselli, P.S. (1992). Atividade ovariana e comportamento reprodutivo no periodo pos-parto em búfalos (Bubalus bubalis). M.S. Thesis, Faculdade de Medicina Veterinaria e Zootecnia, Universidade de Sao Paulo, (SP) Brazil, 100 pages.

ARTIFICIAL INSEMINATION IN ARGENTINA

TABLE 1 PERCENTAGE OF OCCURRENCE OFHEAT SIGNS IN BUFFALO

FEMALES AS OBSERVED BY DIFFERENT AUTHORS

Heat signs (%) Jacomini Baruselli Vale Crudeli Crudeli

(1989) (1992) (1994) (1996) (1997) Stand to be mounted 100 100 100 41.6 86.1 Vulvar edema 87.5 79.3 67.0 66.6 80.7 Spontaneous mucus discharge 12.9 13.8 76.1 16.6 6.6 Frequent urination 19.4 72.4 67.0 75.0 30.0 Flehemen -

-

-

46.1 10.0 Increased uterine tone 83.9 93.1 - 71.4 95.0 Vulvar hyperemia 96.4 79.3 65.9 85.7 88.9 Mucus discharge at rectal palpation 96.8 69.0 - 66.6 40.0 Homosexual mounts 6.5 3.4 32.9 16.6 9.6

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Crudeli,G.A. Artificial Insemination in Buffalo in Northeastern Argentina. Pág. 6 © Copyright 2003. INTA. EEA Colonia Benítez. Marcos Briolini s/n (3505) Colonia Benítez, Chaco Argentina

Baruselli, P.S. (1994). Sexual behavior in buffaloes. In: Actas del IV Congreso Mundial

de Criadores de Bufalos. Sao Paulo, Brazil, Vol. 1: 158-173.

Buffalo Journal 15 (1)

Baruselli, P.S., Barnabe, V.H., Barnabe, RC., Visintin, J.A and Molero-Fihlo J.R (1994). Artificial Insemination in buffalo. In: Actas del IV Congreso Mundial de

Criadores de Bufalos. Sao Paulo, Brazil Vol. III: 649-651.

Baruselli, P.S., Barnabe, V.H., Barnabe, RC., Visintin, J.A, Molero-Fihlo J.R. BARNABE, RC. and Porto-Fihlo, R (1995). Condiçao Corporal ao Parto e Eficienda Reprodutiva de Femeas Bubalinas Inseminadas Artificialmente. In: Actas del XI Congreso Brasileiro de Reproduçao Animal. Belo Horizonte (MG), Brazil, 380 .

Barnabe, V.H. (1994). Artificial insemination using two different diluents. Buffalo Journal Supplement No. 2: 83-86.

Barnabe, V.H., Baruselli, P.S. and Barnabe, RC. (1995). Implantaçao de programas de inseminaçao artificial. In: Actas del XI Congreso Brasileiro de Reproducao Animal.

Belo Horizonte, (MG), Brazil, 142

Crudeli, G.A, Maldonado, P., Martin, R, Ragazzi, A., Storani, C., Romero, S., Flores, S., Rivero, S. and Bogado, E. (1995). Produccion de Bufalos en el NEA: Una alternativa. In: Actas del JOVECOR, Corrientes, Argentina 8: 17.

Crudeli, G.A, Maldonado Varga S,P., Stahringer, RC., Flores Barbaran, M.S. (1996a). Inseminación Artificial en Bufalos. In: Actas de Reunion de Comunicaciones Cientificas y Tecnológicas de la UNNE. Resistencia. Argentina V. II (4) : 23.

Crudeli, G.A, Maldonado Varga, S. P., Stahringer, R.C. and Flores Barbaran, M.S. (1996b). Evaluacion de Reproductores Bubalinos (machos) en el NEA In: Actas de las XVII Sesiones de Comunicaciones Cientificas, Facultad de Ciencias Veterinarias,

UNNE, Corrientes, Argentina.

Crudeli, G.A., Maldonado Varga, S. P., and Flores Barbaran, M.S. (1996c). Comportamiento Reproductivo del Bufalo en el Nordeste Argentino. In: Actas del XV

Congreso Panamericano de Ciencias Veterinarias, Campo Grande, (MS), Brazil, 388.

Hafez, E.S.E. (1989). Ganado bovino y bufalo de agua. In: Reproduccion e

Inseminacion Artificial en Animales. Nueva Editorial Interamericana SA, 5th, Mexico D.F. (ed).

Hegazy, M.A, Essawy, S.A, El-Wishy, AA and Youssef, AH. (1994). Effect of body condition score on reproductive perfomance of buffaloes. In: Anales del IV Congreso Mundial de Criadores de Bufalos. Sao Paulo, Brazil, Vol. III: 630631.

Jacomini, J.O. (1989). Aspectos do ciclo estral e do comportamento sexual de bufalas (Bubalus bubalis, var. Bubalis). M.S. Thesis, Escola de Veterinaria, Universidad

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Crudeli,G.A. Artificial Insemination in Buffalo in Northeastern Argentina. Pág. 7 © Copyright 2003. INTA. EEA Colonia Benítez. Marcos Briolini s/n (3505) Colonia Benítez, Chaco Argentina Federal de Minas Gerais, (MG), Brazil.

Riveiro, H.F.L., Lau, H.D., Silva, A.O.A, Sousa, J.S. and Vale, W.G. (1994). Preliminary report on artificial insemination in buffaloes of the Amazon region through imported semen. In: Anales del IV Congreso Mundial de Criadores de Bufalos. Sao Paulo, Brazil. Vol. III: 593-593.

Artificial Insemination in Argentina

Vale, W.G. (1988). Bubalinos, fisiologia e patologia da reproducao. Comunicado Tecnico da Fundacao Cargill, Campinas, (SP), Brazil.

Vale, W.G. (1994a). Preparation of a "teaser" buffalo bull for use in artificial insemination. Buffalo Journal 10 (1) : 75-79.

Vale; W.G. (1994b). Reproductive management of water buffalo under Amazon conditions. Buffalo Journal 10 (2): 85-90.

Zava, M. (1992). Servicio natural e inseminación artificial. In: Producción de búfalos.

References

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