Prohibition of hormones in animal reproduction: what to expect and what to do?

As our understanding of ovarian function in cattle has improved, our ability to control it has also increased. The development of Fixed-Time Artificial Insemination (FTAI) protocols at the end of the 20th century has increased exponentially the number of animals inseminated over the last 20 years. The main reasons for this growth were the possibility of obtaining acceptable pregnancy rates without heat detection and, above all, the induction of cyclicity in suckled cows in postpartum anestrus and prepubertal heifers at the beginning of the breeding season. Most FTAI treatments in South America have been based on the use of progesterone (P4) releasing devices and estradiol to synchronize both follicular wave emergence and ovulation, with pregnancy rates ranging from 40 to 60%. These protocols are implemented on a regular basis, allowing producers access to high-quality genetics, and increasing the overall pregnancy rates during the breeding season. In addition, it provided the professionals involved in these programs with a new source of income and the diversification of their practices into activities other than their usual clinical work. Many of these practices are now apparently at risk from restrictions on the use of estradiol by the European Union (EU) and other countries. However, the development of alternative protocols based on GnRH, with P4 devices and eCG and other new products that are not in the market yet will allow us to adapt to the new times that are coming. Logically, the challenge has already been raised and we must learn to use alternative protocols to try to continue increasing the use of this technology in beef and dairy herds. The objective of the present review is to describe the main aspects of banning estradiol in livestock production, the negative impacts on reproductive efficiency, and to present some alternative FTAI protocols for dairy and beef cattle.(AU)

In vitro sperm characteristics and in vivo fertility of sex-sorted and conventional semen in suckled Nelore cows at a traditional schedule for timed-AI with estrus detection / Características espermáticas in vitro e fertilidade in vivo de sêmen convencional e sexado em vacas Nelore pósparto submetidas à IATF tradicional com detecção de estro

The aim of this study was to assess in vitro sperm characteristics and pregnancies/AI (P/AI) of conventional and sex-sorted semen at timed-AI of suckled, multiparous Nelore cows. All cows (n=348) were submitted to a traditional estradiol/progesterone (P4)-based protocol. At 48h after P4-device removal, the estrous behavior was recorded, and AI was performed with conventional or sex-sorted semen from two bulls. The following sperm assessments were performed: CASA, Hyposmotic Test, sperm morphometry and chromatin structure by TB staining. P/AI were reduced (P<0.001) for sex-sorted compared to conventional semen in cows expressing estrus (27vs47%) or not (11vs.37%). Membrane integrity (Bull1: 30.3±9.6 vs. 52.3±12.4%, P=0.01; Bull2: 24.5±3.0 vs. 48.7±1.6%, P=0.006) and sperm concentration (Bulll: 23.2±0.6 vs. 43.0±0.8x10 sperm/mL, P<0.001; Bull2: 25.1+2.8 VS. 42.1±0.7x10 sperm/mL; P<0.001) were reduced in sex-sorted compared to conventional semen, for both bulls. Total and progressive motility were reduced in sex-sorted semen for Bull1 (TM: 49.7±15.9 vs. 94.9±1.9%, P=0.007; PM: 16.7±3.4 vs. 44.1±13.2%, P=0.009) and no differences were detected for Bull2 (TM: 45.0±17.5 vs. 68.2±19.1%, P=0.098; PM: 12.8±4.7 vs. 30.0±13.0%, P=0.065). Sperm ellipticity from sex-sorted was lower than conventional semen for Bull2 (0.306±0.01 vs. 0.342±0.02, P=0.02) and no difference was detected for Bull1 (0.332±0.01 vs. 0.330±0.01, P=0.55). Reduced in vivo fertility was observed for sex-sorted semen, regardless of estrous behavior. In vitro sperm quality of sex-sorted semen was compromised for both bulls, but differently affected for each sire.

O estudo teve como objetivo avaliar características espermáticas in vitro e a taxa de concepção (TC) de sêmen convencional e sexado em um programa de IATF tradicional de vacas Nelore pós-parto. Todas as vacas (n=348) foram submetidas ao mesmo protocolo de IATF à base de estradiol e de progesterona. Após 48 horas da retirada do implante, foi determinada a expressão de estro dos animais e a IA foi realizada com sêmen convencional e sexado de dois touros Angus. As seguintes características espermáticas foram avaliadas: análise computadorizada do sêmen, teste hiposmótico, morfometria espermática e estrutura cromatinica por meio da coloração com azul de toluidina. A TC foi menor (P<0,001) para sêmen sexado comparado ao convencional, em vacas que expressaram estro (27 vs. 47%) e que não apresentaram estro (11 vs. 37%). A integridade da membrana plasmática (Touro 1: 30,319,6 vs. 52,3+12,4%, P=0,010; Touro 2: 24,5+3,0 vs. 48,7±1,6%, P=0,006) e a concentração espermática (Touro 1: 23,2±0,6 vs. 43,0±0,8x10 sperm/mL, P<0,001; Touro 2: 25,1+2,8 vs. 42,1 +0,7x10'sperm/mL, P<0,001) foram menores no sêmen sexado comparado ao convencional, para ambos os touros. Motilidades total e progressiva foram menores no sêmen sexado comparado ao convencional para o Touro 1 (MT: 49,7±15,9 vs. 94,9±1,9%, P=0,007; MP: 16,7+3,4 vs. 44,1+13,2%, P=0,009), enquanto diferenças não foram detectadas no Touro 2 (MT: 45,0±17,5 vs. 68,2±19,1%, P=0,098; MP: 12,8±4,7 vs. 30,0±13,0%, P=0,065). Elipticidade espermática do sêmen sexado foi menor do que do sêmen convencional no Touro 2 (0,306±0,01 vs. 0,342±0,02, P=0,020), mas não houve diferença no Touro 1 (0,332±0,01 vs. 0,330±0,01, P=0,552). Reduzida fertilidade in vivo foi observada para o sêmen sexado em relação ao convencional, independentemente da expressão de cio das vacas. A qualidade seminal in vitro do sêmen sexado foi comprometida para ambos os touros, mas diferentemente afetada para cada reprodutor.

Influence of transcervical infusion of seminal plasma on the farrowing rate and litter size in artificially inseminated sows / [Influência da infusão transcervical de plasma seminal na taxa de partos e tamanho da ninhada em porcos inseminados artificialmente]

Recent studies have focused on the use of seminal plasma to increase sow fertility after classical intracervical artificial insemination (AI). The aim of the present study was to investigate the influence of seminal plasma infusion, prior to the application of conventional AI dose, on the fertility rate in sows. A total of 114 sows were treated with intrauterine infusion of 30ml seminal plasma (SP), while 114 control sows were infused by physiological solution (PS), immediately before the application of conventional AI dose. The experiment was conducted at one commercial pig farm in Serbia, which is comprised of 1,500 sows in the breeding herd. Intrauterine infusion of seminal plasma produced significantly (P<0.05) higher farrowing rate (93.8%) and significantly (P<0.01) more live-born piglets per litter (12.27), compared with the control sows (83.33% farrowing rate and 10.48 piglets). The present results show that intrauterine infusion of seminal plasma can be a useful tool for increasing the fertility rate in artificially inseminated sows, under the conditions of practical intensive pig production.(AU)

Estudos recentes concentraram no uso de plasma seminal para aumentar a fertilidade de porcos após inseminação artificial intracervical clássica (AI). O objetivo do presente estudo foi investigar a influência da infusão de plasma seminal, antes da aplicação da dose de AI convencional, na taxa de fertilidade de porcas. 114 porcas foram tratadas com infusão intrauterina de 30ml plasma seminal, e 114 porcas de controle receberam infusão de solução fisiológica (PS) imediatamente antes da aplicação da dose convencional de AI. O experimento foi realizado em uma fazenda de porcos comercial na Serbia, que é composta de 1.500 porcas no rebanho de reprodução. A infusão intrauterina de plasma seminal produziu uma taxa de fertilidade (93,8%) significativamente maior (P<0.05), e significativamente mais (P<0.01) leitões nascidos vivos por ninhada (12,27) comparado com as porcas de controle (83,33% taxa de fertilidade e 10,48 leitões). Os resultados mostram que infusão intrauterina com plasma seminal pode ser uma ferramenta útil para aumentar a taxa de fertilidade em porcas inseminadas artificialmente, sob as condições de prática de produção intensiva de porcos.(AU)

Influência da dificuldade de inseminação, temperamento e cortisol plasmático sobre a taxa de concepção de vacas e novilhas da raça Nelore inseminadas em tempo fixo / Influence of difficulty of insemination, temperament and plasma cortisol on conception rate of Nelore cows and heifers in timed AI

O objetivo do presente experimento foi avaliar efeito do estresse e da dificuldade de inseminação (DifIA) sobre a taxa de concepção (TC) de vacas (n=93) e novilhas (n= 72) Nelore submetidas à IATF. No D9, anotou-se nota de temperamento (NTe) e tempo da saída do brete (TSB) de todos os animais e coletou-se sangue das novilhas. No dia da IATF (D11), anotou-se NTe, TSB, DifIA e tempo de IA. A TC foi 36% para vacas e 46% para novilhas (P>0,05). Não foi observado efeito de Nte sobre TC (P>0,05). Porém, houve tendência para maior (P<0,10) TC nos animais que não apresentaram dificuldade de inseminação (DifIA1; TP=42%) em comparação aos animais com moderada ou alta dificuldade (DifIA2+DifIA3; TP=27%). Foi observado menor (P<0,05) tempo de IA para animais DifIA1 (17:31±06:02s) que animais DifIA2-3 (30:10±15:45s). Novilhas com maiores (P<0,05) níveis de cortisol apresentaram maior NTe (P<0,05). Entretanto, TC (59%) das novilhas menos reativas (cortisol=4,12±1,12ng/mL; NTe=3,2±0,6) não diferiu da TC (41%; P>0,05) das mais agitadas (cortisol=7,76±1,33ng/mL; NTe=3,82±0,79). Concluiu-se que avaliações de temperamento se relacionaram com nível de estresse, embora esses parâmetros não tenham afetado a TC deste trabalho. A maior dificuldade e/ou tempo necessário para se completar a IA demonstrou ser um potencial fator para a redução da fertilidade na IATF.(AU)

The objective was to evaluate the influence of stress and difficulty of insemination (DifIA) on conception rate (CR) of Nellore cows (n= 93) and heifers (n= 72) in Timed-AI. On D9, temperament (NTe) and time for chute exit (TSB) were recorded for all animals, and blood samples were colected from heifers. On the day of Timed-AI (D11), NTe, TSB, DifIA and time for AI were recorded. For cows, CR was 36% and for heifers 46% (P> 0.05). No effect (P> 0.05) of NTe was observed on CR. However, a tendency (P< 0,10) for higher CR was observed in animals with no difficulty for insemination (DifIA1; CR=42%) compared to animals that presented moderate or high difficulty (DifIA2+DifIA3, CR=27%). Time required for AI was lower (P< 0.05) in animals DifIA1 (17:31±06:02sec) than in animals DifIA2-3 (30:10±15:45sec). Heifers with greater (P< 0.05) cortisol levels presented higher Nte (P< 0.05). However, CR (59%) of less reactive heifers (cortisol=4,12±1,12ng/mL; NTe=3,2±0,6) did not differ from CR (41%; P> 0.05) of stressed animals (cortisol=7,76±1,33ng/mL; NTe=3,82±0,79). It was concluded that assessments of temperament were related to stress level, although these parameters did not affect the CR of this study. However, the higher difficulty and/or time to complete AI showed to be a potential factor for reducing fertility after timed-AI.(AU)

Influência da dificuldade de inseminação, temperamento e cortisol plasmático sobre a taxa de concepção de vacas e novilhas da raça Nelore inseminadas em tempo fixo / Influence of difficulty of insemination, temperament and plasma cortisol on conception rate of Nelore cows and heifers in timed AI

O objetivo do presente experimento foi avaliar efeito do estresse e da dificuldade de inseminação (DifIA) sobre a taxa de concepção (TC) de vacas (n=93) e novilhas (n= 72) Nelore submetidas à IATF. No D9, anotou-se nota de temperamento (NTe) e tempo da saída do brete (TSB) de todos os animais e coletou-se sangue das novilhas. No dia da IATF (D11), anotou-se NTe, TSB, DifIA e tempo de IA. A TC foi 36% para vacas e 46% para novilhas (P>0,05). Não foi observado efeito de Nte sobre TC (P>0,05). Porém, houve tendência para maior (P<0,10) TC nos animais que não apresentaram dificuldade de inseminação (DifIA1; TP=42%) em comparação aos animais com moderada ou alta dificuldade (DifIA2+DifIA3; TP=27%). Foi observado menor (P<0,05) tempo de IA para animais DifIA1 (17:31±06:02s) que animais DifIA2-3 (30:10±15:45s). Novilhas com maiores (P<0,05) níveis de cortisol apresentaram maior NTe (P<0,05). Entretanto, TC (59%) das novilhas menos reativas (cortisol=4,12±1,12ng/mL; NTe=3,2±0,6) não diferiu da TC (41%; P>0,05) das mais agitadas (cortisol=7,76±1,33ng/mL; NTe=3,82±0,79). Concluiu-se que avaliações de temperamento se relacionaram com nível de estresse, embora esses parâmetros não tenham afetado a TC deste trabalho. A maior dificuldade e/ou tempo necessário para se completar a IA demonstrou ser um potencial fator para a redução da fertilidade na IATF.(AU)

The objective was to evaluate the influence of stress and difficulty of insemination (DifIA) on conception rate (CR) of Nellore cows (n= 93) and heifers (n= 72) in Timed-AI. On D9, temperament (NTe) and time for chute exit (TSB) were recorded for all animals, and blood samples were colected from heifers. On the day of Timed-AI (D11), NTe, TSB, DifIA and time for AI were recorded. For cows, CR was 36% and for heifers 46% (P> 0.05). No effect (P> 0.05) of NTe was observed on CR. However, a tendency (P< 0,10) for higher CR was observed in animals with no difficulty for insemination (DifIA1; CR=42%) compared to animals that presented moderate or high difficulty (DifIA2+DifIA3, CR=27%). Time required for AI was lower (P< 0.05) in animals DifIA1 (17:31±06:02sec) than in animals DifIA2-3 (30:10±15:45sec). Heifers with greater (P< 0.05) cortisol levels presented higher Nte (P< 0.05). However, CR (59%) of less reactive heifers (cortisol=4,12±1,12ng/mL; NTe=3,2±0,6) did not differ from CR (41%; P> 0.05) of stressed animals (cortisol=7,76±1,33ng/mL; NTe=3,82±0,79). It was concluded that assessments of temperament were related to stress level, although these parameters did not affect the CR of this study. However, the higher difficulty and/or time to complete AI showed to be a potential factor for reducing fertility after timed-AI.(AU)

Evolution of knowledge on ovarian physiology and its contribution to the widespread application of reproductive biotechnologies in South American cattle

As our understanding of ovarian function in cattle has improved, our ability to control it has also increased. Luteal function in cattle has been studied in detail, and prostaglandin F2α has been used for several years for the elective induction of luteal regression. More recently, follicle wave dynamics has been studied and protocols designed to induce follicular wave emergence and ovulation have reduced, and even eliminated, the need for estrus detection. The addition of progestin-releasing devices, estradiol, GnRH and equine chorionic gonadotropin (eCG) have provided opportunities for fixed-time AI (FTAI) and possibilities for increased pregnancy rates. In embryo transfer programs, these same treatments have eliminated the need for estrus detection, permitting fixed-time embryo transfer and the initiation of superstimulatory treatments without regard to day of the estrous cycle. Collectively, new treatment protocols have facilitated the application of assisted reproductive technologies, and this is especially true in South America. Over the last 20 years, the use of AI in South America has increased, due largely to the use of FTAI. There has been more than a 10-fold increase in the use of FTAI in Brazil with more than 11 million treatments in 2016, representing 85% of all AI. Similar trends are occurring in Argentina and Uruguay. Production of in vivo-derived (IVD) embryos has remained relatively stable over the years, but in vitro embryo production (IVP) has increased dramatically over the past 10 to 15 years, especially in Brazil where more than 300,000 IVP embryos were produced in 2010. World-wide, more than 666,000 bovine IVP embryos were produced in 2016, of which more than 57% were produced in South America. The use of assisted reproductive technologies has facilitated the dissemination of improved genetics and increased reproductive performance; other South American countries are now following suit.(AU)

Evolution of knowledge on ovarian physiology and its contribution to the widespread application of reproductive biotechnologies in South American cattle

As our understanding of ovarian function in cattle has improved, our ability to control it has also increased. Luteal function in cattle has been studied in detail, and prostaglandin F2α has been used for several years for the elective induction of luteal regression. More recently, follicle wave dynamics has been studied and protocols designed to induce follicular wave emergence and ovulation have reduced, and even eliminated, the need for estrus detection. The addition of progestin-releasing devices, estradiol, GnRH and equine chorionic gonadotropin (eCG) have provided opportunities for fixed-time AI (FTAI) and possibilities for increased pregnancy rates. In embryo transfer programs, these same treatments have eliminated the need for estrus detection, permitting fixed-time embryo transfer and the initiation of superstimulatory treatments without regard to day of the estrous cycle. Collectively, new treatment protocols have facilitated the application of assisted reproductive technologies, and this is especially true in South America. Over the last 20 years, the use of AI in South America has increased, due largely to the use of FTAI. There has been more than a 10-fold increase in the use of FTAI in Brazil with more than 11 million treatments in 2016, representing 85% of all AI. Similar trends are occurring in Argentina and Uruguay. Production of in vivo-derived (IVD) embryos has remained relatively stable over the years, but in vitro embryo production (IVP) has increased dramatically over the past 10 to 15 years, especially in Brazil where more than 300,000 IVP embryos were produced in 2010. World-wide, more than 666,000 bovine IVP embryos were produced in 2016, of which more than 57% were produced in South America. The use of assisted reproductive technologies has facilitated the dissemination of improved genetics and increased reproductive performance; other South American countries are now following suit.

Effect of timing of artificial insemination in relation to onset of standing estrus on pregnancy per AI in Nili-Ravi buffalo

The aim of the present study was to determine the optimum time of artificial insemination after the beginning of standing estrus in buffalo. Nili-Ravi buffalo (n = 109) during breeding season were exposed to teaser bull at 12 hours interval to determine the standing heat (0 h). Buffalo were randomly allocated to different time groups and a single artificial insemination was performed either at 0 h (n = 30), 12 h (n = 27), 24 h (n = 28) or 36 h (n = 24). In a subset of buffalo (n = 38) ultrasonography was performed, twice daily from 0 h (onset of standing heat) to determine the time of ovulation. Pregnancy diagnosis was performed 35-40 days after AI. Results revealed that mean time of ovulation from onset of standing heat was 34.7 ± 0.96 h (range 30 to 42 h). Higher (P < 0.05) pregnancy per AI were achieved in buffalo when inseminated at 24 h (15/28; 53%) compared to 0 h (8/30; 26%) and 36 h (3/24; 13%). Pregnancy per AI, was in-between, in buffalo, inseminated at 12 h (10/27; 37%) and did not differ (P > 0.05) with those bred either at 24 h or 0 h. The odds ratio further confirmed that the occurrence of pregnancy per AI was two times higher in buffalo inseminated at 24 h as compared to those at 12 h. It is concluded that optimal pregnancy per AI can be achieved when buffalo are bred artificially 24 h after the onset of standing heat.(AU)

Effect of timing of artificial insemination in relation to onset of standing estrus on pregnancy per AI in Nili-Ravi buffalo

The aim of the present study was to determine the optimum time of artificial insemination after the beginning of standing estrus in buffalo. Nili-Ravi buffalo (n = 109) during breeding season were exposed to teaser bull at 12 hours interval to determine the standing heat (0 h). Buffalo were randomly allocated to different time groups and a single artificial insemination was performed either at 0 h (n = 30), 12 h (n = 27), 24 h (n = 28) or 36 h (n = 24). In a subset of buffalo (n = 38) ultrasonography was performed, twice daily from 0 h (onset of standing heat) to determine the time of ovulation. Pregnancy diagnosis was performed 35-40 days after AI. Results revealed that mean time of ovulation from onset of standing heat was 34.7 ± 0.96 h (range 30 to 42 h). Higher (P 0.05) with those bred either at 24 h or 0 h. The odds ratio further confirmed that the occurrence of pregnancy per AI was two times higher in buffalo inseminated at 24 h as compared to those at 12 h. It is concluded that optimal pregnancy per AI can be achieved when buffalo are bred artificially 24 h after the onset of standing heat.

Quorum sensing in Enterococcus faecium, Enterococcus faecalis and Bacillus cereus strains isolated from ricotta processing / Quorum sensing em cepas de Enterococcus faecium, Enterococcus faecalis e Bacillus cereus isoladas do processamento de ricota

The quorum sensing phenomenon is a process of intra- and inter-species microbial communication involving the production and detection of extracellular signaling molecules. The autoinducer AI-2 has been proposed to serve as a universal signal for interspecies communication. This study aimed to evaluate the capability of Enterococcus faecium, Enterococcus faecalis, and Bacillus cereus strains isolated from ricotta processing to produce quorum sensing signalling molecules (AI-2). The strains were evaluated for the presence of the luxS gene using the polymerase chain reaction. AI-2 quorum sensing signalling molecules were measured in relative light units (RLUs) using a luminometer. A total of 74% of E. faecium, 91% of E. faecalis, and 95% of B. cereus isolates were positive for luxS gene. In addition, the induced bioluminescence in Vibrio harveyi BB170 was observed in all strains, indicating the presence of the AI-2 autoinducer.(AU)

O fenômeno quorum sensing corresponde a um processo de comunicação intra e interespécies microbianas e é mediado por sinais químicos extracelulares, denominados moléculas sinalizadoras ou auto indutoras (AI). A molécula AI2 está envolvida na comunicação interespécies, denominada sistema universal de comunicação. Este estudo teve como objetivo avaliar a capacidade de Enterococcus faecium, Enterococcus faecalis e Bacillus cereus isolados do processamento de ricota em produzir moléculas sinalizadoras de Quorum sensing (AI-2). Os isolados foram avaliados quanto à presença do gene luxS utilizando a reação em cadeia da polimerase (PCR). As moléculas sinalizadoras (AI-2) foram medidas em unidades relativas de luz (RLU) através de um luminômetro. Um total de 74% dos isolados de E. faecium, 91% de E. faecalis e 95% de B. cereus foram positivos para o gene luxS. Além disso, todos os isolados apresentaram capacidade de induzir o fenômeno de bioluminescência em Vibrio harveyi BB170, indicando a presença de auto indutores AI-2.(AU)

Influence of follicular growth stimulation on fertility of beef cows treated with eCG or FSH in FTAI protocols / Influência da estimulação do crescimento folicular na fertilidade de vacas de corte tratadas com eCG ou FSH em protocolos de IATF

Background: The pregnancy rate in fixed-time insemination (FTAI) programs in beef cattle is affected by several factors, including the size of ovulatory follicle (OF). Induced ovulation of small OF (≤11 mm) reduces the chance of pregnancy after FTAI when compared to ovulation of larger follicles (≥12 mm). Hormonal treatments to enhance the follicle growth have been widely used in comercial beef cattle systems, aiming to improve the development of a dominant follicle. The objective of this experiment was to evaluate the effect of treatment with eCG or FSH at the time of progesterone implant removal on the diameter of OF, final follicle growth and pregnancy rate in beef cows.Materials, Methods & Results: Three hundred and seven suckled beef cows with body condition score 2.80 ± 0.24 (1 to 5 scale) and 45 to 70 days postpartum were synchronized to FTAI. On day 0, intravaginal implants containing 1 g of progesterone, were inserted and administered 2 mg of estradiol benzoate (EB), i.m. On day 7, intravaginal implants were removed and 150 µg of D-cloprostenol were administered i.m. At this time, three groups were assigned according to the treatments: eCG (400 UI eCG, i.m.), FSH (25 UI FSH, i.m.) and Control (1 mL Saline, i.m.). On day 8, all cows were injected with 1mg EB i.m. and were inseminated 52 to 56 h after the implant removal. Transrectal ultrasonography exams of ovaries were performed at the time of implant removal and before FTAI to determine the diameter of the largest follicle and the follicular daily growth. Pregnancy exams were conducted by transretal ultrasonography 30 days after FTAI. Statistical analyzes were performed by Chi-square test for pregnancy rate. Ovarian diameters and follicular daily growth were analyzed by ANOVA with means compared by Tukey test considering significant P value < 0.05.[...]

Influence of follicular growth stimulation on fertility of beef cows treated with eCG or FSH in FTAI protocols / Influência da estimulação do crescimento folicular na fertilidade de vacas de corte tratadas com eCG ou FSH em protocolos de IATF

Background: The pregnancy rate in fixed-time insemination (FTAI) programs in beef cattle is affected by several factors, including the size of ovulatory follicle (OF). Induced ovulation of small OF (≤11 mm) reduces the chance of pregnancy after FTAI when compared to ovulation of larger follicles (≥12 mm). Hormonal treatments to enhance the follicle growth have been widely used in comercial beef cattle systems, aiming to improve the development of a dominant follicle. The objective of this experiment was to evaluate the effect of treatment with eCG or FSH at the time of progesterone implant removal on the diameter of OF, final follicle growth and pregnancy rate in beef cows.Materials, Methods & Results: Three hundred and seven suckled beef cows with body condition score 2.80 ± 0.24 (1 to 5 scale) and 45 to 70 days postpartum were synchronized to FTAI. On day 0, intravaginal implants containing 1 g of progesterone, were inserted and administered 2 mg of estradiol benzoate (EB), i.m. On day 7, intravaginal implants were removed and 150 µg of D-cloprostenol were administered i.m. At this time, three groups were assigned according to the treatments: eCG (400 UI eCG, i.m.), FSH (25 UI FSH, i.m.) and Control (1 mL Saline, i.m.). On day 8, all cows were injected with 1mg EB i.m. and were inseminated 52 to 56 h after the implant removal. Transrectal ultrasonography exams of ovaries were performed at the time of implant removal and before FTAI to determine the diameter of the largest follicle and the follicular daily growth. Pregnancy exams were conducted by transretal ultrasonography 30 days after FTAI. Statistical analyzes were performed by Chi-square test for pregnancy rate. Ovarian diameters and follicular daily growth were analyzed by ANOVA with means compared by Tukey test considering significant P value < 0.05.[...](AU)

Efficiency of different methods of estrus synchronization followed by fixed time artificial insemination in Persian downy does

For evaluating different methods of long term estrous synchronization followed by fixed time artificial insemination and to select the most efficient method, during the breeding season 160 Persian downy does were equally allocated to groups (n = 20/group). Estrus Was synchronized using controlled internal drug release devices alone (CIDR) or with equine chorionic gonadotropin (CIDR-eCG), intravaginal sponge impregnated with 45 mg fluorgestone acetate alone (Sponge) or with eCG (Sponge-eCG),subcutaneous auricular implant of 2 mg norgestomet alone (Implant) or with eCG (Implant-eCG) or two intramuscular injections of 10 mg prostaglandin F2α10 days apart alone (PGF) or accompany with eCG (PGF-eCG). The dose of eCG was 400 IU and injected intramuscularlly at the end of treatments. Heat detection and fixed time artificial insemination were done 12 and 4 hours after the end of the treatments, respectively. The estrus was detected in 94.7-100% of the does who received different forms of progestagens and estrus response was not affected by eCG. Estrus response in PGF-based groups was significantly lower (P <0.05) than progesterone-based groups. Injection of eCG did not affect fertility, fecundity and prolificacy rates in CIDR, Sponge and Implant groups. It also did not affectparturition rate in implant group. Finally, with respect to estrus response, fertility rate, costs and the simplicity of methods it can be concluded that the use of intravaginal CIDR For 16 days without administration of eCG may be a more efficient method for estrus synchronization in fixed-time artificial insemination programs in Persian downy does.

Efficiency of different methods of estrus synchronization followed by fixed time artificial insemination in Persian downy does

For evaluating different methods of long term estrous synchronization followed by fixed time artificial insemination and to select the most efficient method, during the breeding season 160 Persian downy does were equally allocated to groups (n = 20/group). Estrus Was synchronized using controlled internal drug release devices alone (CIDR) or with equine chorionic gonadotropin (CIDR-eCG), intravaginal sponge impregnated with 45 mg fluorgestone acetate alone (Sponge) or with eCG (Sponge-eCG),subcutaneous auricular implant of 2 mg norgestomet alone (Implant) or with eCG (Implant-eCG) or two intramuscular injections of 10 mg prostaglandin F2α10 days apart alone (PGF) or accompany with eCG (PGF-eCG). The dose of eCG was 400 IU and injected intramuscularlly at the end of treatments. Heat detection and fixed time artificial insemination were done 12 and 4 hours after the end of the treatments, respectively. The estrus was detected in 94.7-100% of the does who received different forms of progestagens and estrus response was not affected by eCG. Estrus response in PGF-based groups was significantly lower (P <0.05) than progesterone-based groups. Injection of eCG did not affect fertility, fecundity and prolificacy rates in CIDR, Sponge and Implant groups. It also did not affectparturition rate in implant group. Finally, with respect to estrus response, fertility rate, costs and the simplicity of methods it can be concluded that the use of intravaginal CIDR For 16 days without administration of eCG may be a more efficient method for estrus synchronization in fixed-time artificial insemination programs in Persian downy does.(AU)

Assessment of different in vitro sperm challenges and in vivo fertility of bovine semen batches / Avaliação de diferentes desafios espermáticos in vitro e da fertilidade in vivo de partidas de sêmen bovino

The aim of this work was to submit sperm cells to different laboratory challenges and to compare in vitro results with in vivo semen fertility. Four different batches from the same Brangus bull were used in a timed-AI program of 332 Brangus cows. Each batch (B) was submitted to the following procedure: semen sample was thawed at 36°C for 30 seconds (control). Sperm motility parameters, plasma membrane integrity, sperm morphology, and concentration were assessed. Then, an aliquot of thawed sample was incubated in a water bath at 45°C for 40 min (thermal challenge group; TCG) and another aliquot was centrifuged at 500 xg (Percoll gradient 45%/90%) for 15 min (centrifugation challenge group; CCG). Centrifuged semen was also submitted to another thermal challenge, being incubated (water bath) at 45°C for 40 min (centrifugation + thermal challenge group; CTCG). At the end of each challenge (CCG, TCG, and CTCG), the same laboratory tests used for control group were repeated. The following conception rates (CR) were observed for each batch: B1 = 48.9% (44/90); B2 = 44.2% (23/52); B3 = 55.5% (40/72); B4 = 43.2% (51/118); (p < 0.10). In the lab, B3 presented higher (p ≤ 0.05) progressive motility (PM) than B4 after thawing (control group) and after all sperm challenges (TCG, CCG, and CTCG). However, despite B3 and B4 having demonstrated a similar percentage of plasma membrane integrity (PMI) to the control group (B3 = 66.7 ± 1.3 and B4 = 65.2 ± 3.3), B3 demonstrated higher (P ≤ 0.05) percentage of PMI (37.2 ± 2.5) than B4 (26.7 ± 3.3) after passing through the most stressing in vitro challenge (CTCG). The semen batch presenting the highest resistance to in vitro challenges was the one that presented a trend for higher in vivo fertility, suggesting that submitting semen samples to laboratory challenges may be an interesting alternative for selecting batches with greater field fertility.(AU)

O objetivo deste estudo foi estressar células espermáticas em diferentes desafios laboratoriais e comparar os resultados in vitro com a fertilidade in vivo do sêmen. Quatro partidas de um mesmo touro Brangus foram utilizadas em um programa de IATF de 332 vacas Brangus. Cada partida foi submetida ao seguinte procedimento: a amostra de sêmen foi descongelada a 36°C por 30 segundos (grupo controle). Foram avaliados parâmetros de motilidade espermática (CASA), integridade da membrana plasmática (PMI), morfologia e concentração espermática. Em seguida, uma alíquota da amostra descongelada foi incubada em banho-maria a 45°C durante 40 minutos (grupo de desafio térmico, TCG) e outra alíquota foi centrifugada a 500 xg (gradiente de Percoll 45%/90%) durante 15 min (grupo desafio de centrifugação, CCG). Uma aliquota do sêmen centrifugado foi ainda submetida ao desafio térmico, sendo incubado a 45°C durante 40 min (grupo de desafio térmico + centrifugação, CTCG). No final de cada desafio (CCG, TCG e CTCG), os mesmos testes laboratoriais utilizados para o grupo de controle foram realizados. A seguinte taxa de concepção (CR) foi observada para cada partida (B): B1 = 48,9% (44/90), B2 = 44,2% (23/52), B3 = 55,5% (40/72) e B4 = 43,2% (51/118); (P < 0,10). No laboratório, B3 apresentou maior (P ≤ 0,05) motilidade progressiva (PM) do que B4 logo após o descongelamento (grupo controle) e após todos os desafios laboratoriais (TCG, CCG e CTCG). Porém, apesar de B3 e B4 demonstrarem similar porcentagem de PMI no grupo controle (B3 = 66,7 ± 1,3 e B4 = 65,2 ± 3,3), B3 apresentou maior (P ≤ 0,05) PMI (37,2 ± 2,5%) do que B4 (26,7 ± 3,3%) após passar pelo maior desafio laboratorial (CTCG). A partida seminal que in vitro apresentou maior resistência aos desafios laboratoriais foi a mesma que apresentou tendência para maior fertilidade in vivo. Assim, sugere-se que submeter amostras seminais a desafios laboratoriais pode ser uma alternativa interessante para selecionar partidas com maior fertilidade a campo.(AU)

Assessment of different in vitro sperm challenges and in vivo fertility of bovine semen batches / Avaliação de diferentes desafios espermáticos in vitro e da fertilidade in vivo de partidas de sêmen bovino

The aim of this work was to submit sperm cells to different laboratory challenges and to compare in vitro results with in vivo semen fertility. Four different batches from the same Brangus bull were used in a timed-AI program of 332 Brangus cows. Each batch (B) was submitted to the following procedure: semen sample was thawed at 36°C for 30 seconds (control). Sperm motility parameters, plasma membrane integrity, sperm morphology, and concentration were assessed. Then, an aliquot of thawed sample was incubated in a water bath at 45°C for 40 min (thermal challenge group; TCG) and another aliquot was centrifuged at 500 xg (Percoll gradient 45%/90%) for 15 min (centrifugation challenge group; CCG). Centrifuged semen was also submitted to another thermal challenge, being incubated (water bath) at 45°C for 40 min (centrifugation + thermal challenge group; CTCG). At the end of each challenge (CCG, TCG, and CTCG), the same laboratory tests used for control group were repeated. The following conception rates (CR) were observed for each batch: B1 = 48.9% (44/90); B2 = 44.2% (23/52); B3 = 55.5% (40/72); B4 = 43.2% (51/118); (p < 0.10). In the lab, B3 presented higher (p ≤ 0.05) progressive motility (PM) than B4 after thawing (control group) and after all sperm challenges (TCG, CCG, and CTCG). However, despite B3 and B4 having demonstrated a similar percentage of plasma membrane integrity (PMI) to the control group (B3 = 66.7 ± 1.3 and B4 = 65.2 ± 3.3), B3 demonstrated higher (P ≤ 0.05) percentage of PMI (37.2 ± 2.5) than B4 (26.7 ± 3.3) after passing through the most stressing in vitro challenge (CTCG). The semen batch presenting the highest resistance to in vitro challenges was the one that presented a trend for higher in vivo fertility, suggesting that submitting semen samples to laboratory challenges may be an interesting alternative for selecting batches with greater field fertility.(AU)

O objetivo deste estudo foi estressar células espermáticas em diferentes desafios laboratoriais e comparar os resultados in vitro com a fertilidade in vivo do sêmen. Quatro partidas de um mesmo touro Brangus foram utilizadas em um programa de IATF de 332 vacas Brangus. Cada partida foi submetida ao seguinte procedimento: a amostra de sêmen foi descongelada a 36°C por 30 segundos (grupo controle). Foram avaliados parâmetros de motilidade espermática (CASA), integridade da membrana plasmática (PMI), morfologia e concentração espermática. Em seguida, uma alíquota da amostra descongelada foi incubada em banho-maria a 45°C durante 40 minutos (grupo de desafio térmico, TCG) e outra alíquota foi centrifugada a 500 xg (gradiente de Percoll 45%/90%) durante 15 min (grupo desafio de centrifugação, CCG). Uma aliquota do sêmen centrifugado foi ainda submetida ao desafio térmico, sendo incubado a 45°C durante 40 min (grupo de desafio térmico + centrifugação, CTCG). No final de cada desafio (CCG, TCG e CTCG), os mesmos testes laboratoriais utilizados para o grupo de controle foram realizados. A seguinte taxa de concepção (CR) foi observada para cada partida (B): B1 = 48,9% (44/90), B2 = 44,2% (23/52), B3 = 55,5% (40/72) e B4 = 43,2% (51/118); (P < 0,10). No laboratório, B3 apresentou maior (P ≤ 0,05) motilidade progressiva (PM) do que B4 logo após o descongelamento (grupo controle) e após todos os desafios laboratoriais (TCG, CCG e CTCG). Porém, apesar de B3 e B4 demonstrarem similar porcentagem de PMI no grupo controle (B3 = 66,7 ± 1,3 e B4 = 65,2 ± 3,3), B3 apresentou maior (P ≤ 0,05) PMI (37,2 ± 2,5%) do que B4 (26,7 ± 3,3%) após passar pelo maior desafio laboratorial (CTCG). A partida seminal que in vitro apresentou maior resistência aos desafios laboratoriais foi a mesma que apresentou tendência para maior fertilidade in vivo. Assim, sugere-se que submeter amostras seminais a desafios laboratoriais pode ser uma alternativa interessante para selecionar partidas com maior fertilidade a campo.(AU)

Analysis of lower urinary tract disease of dogs / Análise de doenças do trato urinário inferior em cães

Lower urinary tract diseases (LUTD) include different conditions that affect the urinary bladder, urethra and prostate. The objective of this study was to determine the frequency of different related diseases, to characterize the population affected, and to determine risk factors in dogs. The clinical cases were diagnosed with LUTD through physical examination, and clinical laboratory and imaging studies. Male dogs had a greater predisposition to present a LUTD. Dogs from 3 months to 18 years with a median of 8 years were affected, and the most affected breeds were Poodle, Labrador, German shepherd, Schnauzer, Cocker Spaniel and Chihuahua. The LUTD presented with the following frequencies: bacterial urinary tract infection 34.02%; micturition disorders 22.68%; urolithiasis 20.61%; prostatic disease 14.43%; traumatic problems 8.24%. Sixty-seven per cent of the cases were specific diseases, such as uncomplicated and complicated bacterial urinary tract infections, urinary retention of neurologic origin and silica urolithiasis.(AU)

Doenças do trato urinário inferior (DTUI) incluem várias condições clínicas que afetam a bexiga, próstata e uretra. O objetivo deste estudo foi determinar a frequência de diferentes doenças relacionadas, caracterizar a população afetada, e determinar os fatores de risco em cães. Os casos clínicos com DTUI foram diagnosticados através de exame físico, estudos laboratoriais e de imagem. Os cães machos apresentam uma maior predisposição para DTUI. Foram afectados cães com idades compreendidas entre os 3 meses a 18 anos, com uma mediana de oito anos, sendo as raças mais predispostas os Poodle, Labrador, Pastor Alemão, Schnauzer, Cocker Spaniel e Chihuahua. A etiologia DTUI apresentou as seguintes frequências: 34,02% foram originadas em infecção do trato urinário inferior; 22,68% em distúrbios miccionais; 20,61% em urolitíase; 14,43% em doença prostática; 8,24% em problemas traumáticos. Sessenta e sete por cento dos casos de DTUI estavam associados a doenças específicas, tais como infecções bacterianas do trato urinário não complicadas e complicadas, retenção urinária neurogénica e urolitíase por sílica.(AU)

Effect of GnRH or estradiol benzoate on reproductive traits during a Heatsynch protocol in dairy cows

Background: Milk production of cows is closely correlated with its reproductive efficiency. One of the several factors influencing the dairy efficiency is the decline in fertility of the cows/heifers. Use of hormonal protocol are often used to improve the fertility of the recipients. Several programs are available to inseminate dairy cows, with variation in the use of different types of hormones, time of use and range of duration period, as well as the time of AI. The objective of this study was to evaluate the efficiency of follicular wave emergence induction of estradiol benzoate in comparison to GnRH in a Heatsynch protocol and its influence on reproductive parameters. Materials, Methods & Results: In this study were used primiparous and multiparous Holstein cows (n = 325). Animals were divided randomly into two different treatments according with the hormone used to induce follicular wave emergence (D0). The GnRH group (n = 167), was given 25 mg, while in the EB group (n = 158) was used 2 mg. At same day, were inserted, a controlled internal drug-releasing containing 1.9 g of progesterone CIDR®. On day 7, CIDR was removed and cows were given 25 mg luteinizing hormone followed by an injection of 1 mg Inducer Ovulation at day 8. Cows were then observed for signs of estrus for at least two hours (twice daily at 7 am and 6 pm) for three days following CIDR® removal. Cows [...](AU)

Effect of GnRH or estradiol benzoate on reproductive traits during a Heatsynch protocol in dairy cows

Background: Milk production of cows is closely correlated with its reproductive efficiency. One of the several factors influencing the dairy efficiency is the decline in fertility of the cows/heifers. Use of hormonal protocol are often used to improve the fertility of the recipients. Several programs are available to inseminate dairy cows, with variation in the use of different types of hormones, time of use and range of duration period, as well as the time of AI. The objective of this study was to evaluate the efficiency of follicular wave emergence induction of estradiol benzoate in comparison to GnRH in a Heatsynch protocol and its influence on reproductive parameters. Materials, Methods & Results: In this study were used primiparous and multiparous Holstein cows (n = 325). Animals were divided randomly into two different treatments according with the hormone used to induce follicular wave emergence (D0). The GnRH group (n = 167), was given 25 mg, while in the EB group (n = 158) was used 2 mg. At same day, were inserted, a controlled internal drug-releasing containing 1.9 g of progesterone CIDR®. On day 7, CIDR was removed and cows were given 25 mg luteinizing hormone followed by an injection of 1 mg Inducer Ovulation at day 8. Cows were then observed for signs of estrus for at least two hours (twice daily at 7 am and 6 pm) for three days following CIDR® removal. Cows [...]

Gonadorelin at the outset and/or end of an ovulation synchronization progesterone estradiol benzoate-based protocol in Nellore females / Gonadorelina no início e/ou no final do protocolo de sincronização da ovulação a base de progesterona e benzoato de estradiol em fêmeas Nelore

This study aimed to evaluate the effects of gonadorelin (GnRH), used both at the outset of a timed artificial insemination (TAI) protocol to synchronize follicular wave recruitment and at the end to synchronize ovulation, on the conception rate (CR) in Nellore cows. The experiment was conducted on two beef cattle farms in the Vale do Araguaia, MT. The body condition score (BCS) evaluation and ultrasound examination to determine ovarian status [follicle diameter < 10 mm, ≥ 10 mm or presence of corpus luteum (CL)] were performed at the beginning (Day 0) of the TAI protocol. Multiparous cows (n = 494) were subjected to the following protocol: Day 0 insertion of intravaginal progesterone device (P4 DIB®, MSD) previously used for 8 or 16 days and IM administration of 2.0 mg of estradiol benzoate (EB, Gonadiol®, MSD); Day 8 P4 device withdrawal, IM injection of 300 IU of equine chorionic gonadotropin (eCG, Folligon®, MSD), 1.0 mg of estradiol cypionate (ECP®, Zoetis) and 0.265 mg of Sodium cloprostenol (PGF2α, Ciosin®, MSD); Day 10 TAI, performed by a single inseminator. Animals were randomly assigned to one of four groups: control (n = 126), GnRH D0 (n = 123), GnRH D10 (n = 123) and GnRH D0 + D10 (n = 122). Animals treated with GnRH received 50 mcg of gonadorelin (GnRH, Cystorelin®, Merial). Pregnancy was diagnosed by ultrasound 39 ± 10 days after TAI. Data were analyzed using the GLIMMIX procedure of SAS program (P < 0.05). The overall CR was 42.71%. Effect of treatment was not detected (P = 0.2482) on CR, which was 36.89% ± 0.19 (47/126) for the control group, 46.08% ± 0.19 (56/123) for GnRH D0, 48.30% ± 0.18 (61/123) for GnRH D10 and 38.48% ± 0.19 (47/122) for GnRH D0 + D10. An effect of farm was detected; the CR was 47.32% ± 0.13 at Farm A and was 38.24% ± 0.13 at Farm B (P = 0.0249). No effects of the interaction between farm and treatment were observed (P = 0.7662), nor of those among ovarian condition and treatment (P = 0.1225) on CR.[...](AU)

Objetivou-se avaliar os efeitos da gonadorelina (GnRH) utilizada para sincronização da emergência da onda de crescimento folicular e para indução da ovulação sincronizada ou em ambos sobre a taxa de concepção (TC) de vacas da raça Nelore. O experimento foi realizado em duas fazendas no Vale do Araguaia, MT. A avaliação do escore de condição corporal (ECC) e a ultrassonografia para determinação da condição dos ovários [folículo com diâmetro < 10 mm, ≥ 10 mm ou presença de corpo lúteo (CL)] foram realizadas no início (Dia 0) do protocolo de inseminação artificial em tempo fixo (IATF). Vacas multíparas (n = 494) foram submetidas ao protocolo: Dia 0 inserção do dispositivo intravaginal de progesterona (P4, DIB®, MSD) utilizado previamente por oito ou 16 dias e aplicação IM de 2,0 mg de Benzoato de Estradiol (BE, Gonadiol®, MSD); Dia 8 remoção do dispositivo, aplicação IM de 300 UI de Gonadotrofina Coriônica Equina (eCG, Folligon®, MSD), de 1,0 mg de Cipionato de Estradiol (ECP®, Zoetis) e de 0,265 mg de Cloprostenol Sódico (PGF2α, Ciosin®, MSD); Dia 10 IATF, realizada por um único inseminador. Os animais foram distribuídos aleatoriamente em quatro grupos: controle (n = 126), GnRH D0 (n = 123), GnRH D10 (n = 123) e GnRH D0 + D10 (n = 122), sendo que os animais tratados com GnRH receberam 50 mcg de gonadorelina (GnRH, Cystorelin®, Merial). O diagnóstico de gestação foi realizado por ultrassonografia 39 ± 10 dias após a IATF. As variáveis foram analisadas pelo procedimento GLIMMIX do programa SAS (P < 0,05). Verificou-se uma TC geral de 42,71%. Não foi detectado efeito dos tratamentos sobre a TC (P = 0,2482), que foi de 36,89% ± 0,19 (47/126) para o grupo controle, 46,08% ± 0,19 (56/123) para GnRH D0, 48,30% ± 0,18 (61/123) para GnRH D10 e de 38,48% ± 0,19 (47/122) para GnRH D0 + D10. Observou-se efeito da variável fazenda, sendo que na Fazenda A a TC foi de 47,32% ± 0,13 e na Fazenda B de 38,24% ± 0,13 (P = 0,0249).[...](AU)