Reproduction technologies in pig farming: artificial insemination - literature review / Tecnologias de reprodução na suinocultura: inseminação artificial - revisão de literatura / Tecnologías de reproducción en la porcicultura: inseminación artificial - revisión de literatura

This summary addresses the use of reproduction technologies in swine farming, with an emphasis on artificial insemination (AI). Brazilian swine farming has been growing significantly and seeks new technologies to achieve high productive indices sustainably and competitively. Pigs present favorable characteristics such as high prolificacy, fertility, rapid growth, feed efficiency, and carcass yield, which has led to intensive development of the activity with advanced genetic selection. AI is widely employed to disseminate genetic material among different regions and farms. Several AI techniques are used in modern swine farming: intrauterine insemination (IUI) allows semen deposition in the uterine region, reducing costs; fixed-time insemination (FTAI) synchronizes estrus in various females, facilitating management and increasing efficiency; deep intrauterine insemination (DIUI) deposits semen in the uterine horns, obtaining better results; and cervical insemination (CI), a traditional technique widely used, although it may be more time-consuming and present higher reflux rates. The success of AI is related to knowledge of the reproductive cycle of sows, proper nutrition, and genetic and environmental factors. Semen quality is essential, requiring collection by trained professionals and evaluation of sperm motility and morphology. Although it is a consolidated technique, there are issues to be further explored to optimize its application, defining the exact moment for insemination, reducing reflux, and adopting effective protocols. AI is an essential tool for the growth of Brazilian swine farming, but it requires continuous studies to maximize its efficiency and results, considering the farm's production goal and the size of the enterprise to achieve high reproductive and productive indices.

Este resumo aborda o uso de tecnologias de reprodução na suinocultura, com ênfase na inseminação artificial (IA). A suinocultura brasileira vem crescendo significativamente e busca novas tecnologias para alcançar altos índices produtivos de maneira sustentável e competitiva. Os suínos apresentam características favoráveis, como alta prolificidade, fertilidade, rápido crescimento, eficiência alimentar e rendimento de carcaça, o que levou ao desenvolvimento intensivo da atividade com seleção genética avançada. A IA é amplamente empregada para disseminar material genético entre diferentes regiões e granjas. Diversas técnicas de IA são utilizadas na suinocultura moderna: a inseminação intrauterina (IAIU) permite a deposição do sêmen na região uterina, reduzindo custos; a inseminação em tempo fixo (IATF) sincroniza o estro em várias fêmeas, facilitando o manejo e aumentando a eficiência; a inseminação intrauterina profunda (IAUP) deposita o sêmen nos cornos uterinos, obtendo melhores resultados; e a inseminação cervical (IAC), técnica tradicional amplamente utilizada, embora possa ser mais demorada e apresentar maiores taxas de refluxo. O sucesso da IA estar relacionado ao conhecimento do ciclo reprodutivo das matrizes, à nutrição adequada e aos fatores genéticos e ambientais. A qualidade do sêmen é essencial, exigindo coleta por profissionais treinados e avaliação da motilidade e morfologia dos espermatozoides. Apesar de ser uma técnica consolidada, há questões a serem aprofundadas para otimizar sua aplicação, definindo o momento exato para a realização da inseminação, a redução do refluxo e adoção de protocolos eficazes. A IA é uma ferramenta essencial para o crescimento da suinocultura brasileira, mas requer estudos contínuos para maximizar sua eficiência e resultados, considerando o objetivo produtivo da granja e o tamanho do empreendimento para alcançar altos índices reprodutivos e produtivos.

Este resumen aborda el uso de tecnologías de reproducción en la producción porcina, con énfasis en la inseminación artificial (IA). La producción porcina brasileña ha crecido significativamente y busca nuevas tecnologías para alcanzar altos índices de productividad de manera sostenible y competitiva. Los cerdos presentan características favorables, como alta prolificidad, fertilidad, rápido crecimiento, eficiencia alimentaria y rendimiento de la canal, lo que ha llevado al desarrollo intensivo de la actividad con selección genética avanzada. La IA se utiliza ampliamente para difundir material genético entre diferentes regiones y granjas. Diversas técnicas de IA son utilizadas en la producción porcina moderna: la inseminación intrauterina (IAIU) permite la deposición del semen en la región uterina, reduciendo costos; la inseminación a tiempo fijo (IATF) sincroniza el estro en varias hembras, facilitando el manejo y aumentando la eficiencia; la inseminación intrauterina profunda (IAUP) deposita el semen en los cuernos uterinos, obteniendo mejores resultados; y la inseminación cervical (IAC), técnica tradicional ampliamente utilizada, aunque puede ser más demorada y presentar mayores tasas de reflujo. El éxito de la IA está relacionado con el conocimiento del ciclo reproductivo de las hembras, la nutrición adecuada y los factores genéticos y ambientales. La calidad del semen es esencial, requiriendo la recolección por profesionales capacitados y la evaluación de la motilidad y morfología de los espermatozoides. A pesar de ser una técnica consolidada, hay aspectos que deben ser profundizados para optimizar su aplicación, como la definición precisa del momento de la inseminación, la reducción del reflujo y la adopción de protocolos eficaces. La IA es una herramienta esencial para el crecimiento de la producción porcina brasileña, pero requiere estudios continuos para maximizar su eficiencia y resultados, considerando el objetivo productivo de la granja y el tamaño del emprendimiento para alcanzar altos índices reproductivos y productivos.

Evaluación de la terapia de reproducción médicamente asistida en 9 años: estudio de cohorte retrospectivo / Medically assisted reproductive therapy assessment in 9 years: retrospective cohort study

OBJETIVO: Describir las tasas de recién nacidos vivos (RNV) y embarazo de la terapia de reproducción médicamente asistida de baja complejidad del Centro de Reproducción Humana de la Universidad de Valparaíso, Chile. MÉTODO: Estudio retrospectivo de todos los ciclos de estimulación ovárica controlada con inseminación intrauterina (IIU) completados, entre los años 2011 y 2019. Se evaluaron las características clínicas basales y los resultados en IIU homólogas y heterólogas según el ciclo inseminado, la causa de infertilidad, el rango etario y el índice de masa corporal (IMC). El desenlace principal fue la tasa de RNV por ciclo inseminado. RESULTADOS: Se estudiaron 1415 ciclos en 700 parejas. La tasa acumulativa de RNV fue del 19,6%, un 18,3% en IIU homóloga y un 39,0% en IIU heteróloga. La tasa de RNV fue del 10,0% al primer ciclo, del 5,8% al segundo ciclo y del 3,7% al tercer o más ciclos. Al separar por IIU heteróloga, esta aumenta al 24,4% al primer ciclo y al 14,6% al segundo ciclo. La tasa de RNV es significativamente mejor en pacientes menores de 35 años (23,7%) y con IMC < 29 (20,8%). CONCLUSIONES: El tratamiento de baja complejidad en pacientes infértiles es una opción terapéutica vigente con una aceptable tasa de RNV por ciclo inseminado. Los resultados están influenciados por la edad y por el IMC.

OBJECTIVE: To describe the rates of live newborns (LNB) and pregnancy of the low complexity therapy of the Centre for Human Reproduction of Universidad de Valparaíso, Chile. METHOD: Retrospective study of all cycles of controlled ovarian stimulation with intrauterine insemination (IUI) completed between 2011-2019. The baseline clinical characteristics and results in homologous and heterologous IUI were evaluated according to inseminated cycle, cause of infertility, age range and body mass index (BMI). The main outcome was rate of LNB per inseminated cycle. RESULTS: 1415 cycles were studied in 700 couples. The cumulative rate of LNB was 19.6%, 18.3% in homologous IUI and 39.0% in heterologous IUI. The LNB rate was 10.0% at the first cycle, 5.8% at the second cycle, 3.7% at the third or more cycles. When separating by heterologous IUI, it increases to 24.4% in the first cycle and 14.6% in the second cycle. The LNB rate is significantly better in patients under 35 years of age (23.7%) and with a BMI less than 29 (20.8%). CONCLUSIONS: Treatment of low complexity in selected infertile patients is a current therapeutic option with an acceptable rate of LNB per inseminated cycle. The results are influenced by age and BMI.

Pre-sincronización con Progesterona para la Inducción de Ciclicidad en Vacas Bos taurus indicus en Periodo de Anestro Posparto / Presynchronization with Progesterone for Induction of Cyclicity in Bos Taurus Indicus Cows in Postpartum Anestrus Period / Pré-sincronização com Progesterona para a Indução de Ciclicidade em Vacas Bos Taurus Indicus em Período de Anestro Pós-Parto

Resumen Objetivo. Este trabajo tuvo como objetivo evaluar el efecto de la pre-sincronización con progesterona (P4) por 3 y 6 días en vacas de raza Nelore en periodo de anestro posparto, sometidas a un programa de inseminación artificial a tiempo fijo (IATF). Materiales y métodos: Fueron utilizadas 74 vacas multíparas de raza Nelore con estado de condición corporal de 2,5. Las vacas fueron sometidas a tres grupos experimentales, grupo control y grupos pre-sincronizados con P4 por un periodo de 3 y 6 días. Para la determinación del estado de anestro posparto, fueron realizadas ecografías y toma de muestras de sangre con la finalidad de verificar la ausencia de P4 endógena. Cuarenta y cinco días después de la IA fue realizado el diagnóstico de la gestación. Los datos fueron obtenidos por análisis descriptivo y en el caso de presentación de diferencias significativas fue utilizado la prueba de Chi-cuadrado, considerándose una frecuencia de P< 0,05. Resultados: Fue observada mayor tasa de gestación para el grupo que fue pre-sincronizado por tres días con P4 (40 %), con relación a los grupos de seis días de pre-sincronización y control (25 % y 32 %, respectivamente). Conclusión: Es posible concluir que la pre-sincronización con P4 por tres días, mejoró la tasa de gestación de vacas de raza Nelore en periodo de anestro posparto.

Abstract Objective: The objective of this study was to evaluate the effect of pre-synchronization with progesterone (P4) for 3 and 6 days in Nelore cows in postpartum anestrus period, submitted to a fixed-time artificial insemination program (FTAI). Materials and methods: Seventy-four multiparous Nelore cows with a body condition of 2.5 were used. The cows were submitted to three experimental groups, control group and groups pre-synchronized with P4 for a period of 3 and 6 days. For the determination of postpartum anestrus status, ultrasound scans and blood samples were taken to verify the absence of endogenous P4. Forty-five days after AI, gestation was diagnosed. Data were obtained by descriptive analysis and in the case of presentation of significant differences the Chi-square test was used, considering a frequency of P < 0.05. Results: A higher gestation rate was observed for the group that was pre-synchronized for three days with P4 (40 %), in relation to the groups of six days of pre-synchronization and control (25 % and 32 %, respectively). Conclusion: It is possible to conclude that the pre-synchronization with P4 for three days improved the gestation rate of Nelore cows in the postpartum anestrus period.

Resumo Objetivo: O objetivo deste estudo foi avaliar o efeito da pré-sincronização com progesterona (P4) durante 3 e 6 dias em vacas de raça Nelore em período de anestro pós-parto, submetidas a um programa de inseminação artificial em tempo fixo (IATF). Materiais e métodos: Foram utilizadas 74 vacas multíparas da raça Nelore com estado de condição corporal de 2,5. As vacas foram submetidas a três grupos experimentais, grupo controle e grupos pré-sincronizados com P4 por um período de 3 e 6 dias. Para a determinação do estado de anestro pós-parto, foram realizadas ultrassonografias e coleta de amostras de sangue com a finalidade de verificar a ausência de P4 endógena. Quarenta e cinco dias após a IA foi realizado o diagnostico da gestação. Os dados foram obtidos por análise descritiva e no caso de diferenças significativas foi utilizado o teste do Quí-quadrado, considerando-se uma freqüência de P < 0,05. Resultados: Foi observada maior taxa de gestação para o grupo que foi pré-sincronizado por três dias com P4 (40 %), com relação aos grupos de seis dias de pré-sincronização e controle (25 % e 32 %, respetivamente). Conclusão: É possível concluir que a pré-sincronização com P4 por três dias, melhorou a taxa de gestação de vacas Nelore em período de anestro pós-parto.

Retirada de corpo estranho (dispositivo impregnado com progesterona) da cavidade abdominal de novilha da raça Jersey ­ relato de caso / Removal of a foreign body (progesterone impregnated device) from the abdominal cavity of a Jersey heifer ­ a case report / Extracción de cuerpo extraño (dispositivo impregnado con progesterona) de la cavidad abdominal de novilla raza Jersey - reporte de caso

A inseminação artificial em tempo fixo (IATF) é uma biotécnica com inúmeras vantagens para a produção de bovinos. Para a realização da IATF, as fêmeas bovinas devem receber um protocolo farmacológico que induz a ovulação de forma sincronizada. Esse protocolo inicia com a inserção de um dispositivo impregnado com progesterona que deve permanecer no fundo vaginal da fêmea por tempo determinado. O presente relato tem por objetivo descrever a remoção de um implante impregnado com progesterona que foi encontrado na cavidade abdominal de uma novilha. Durante um atendimento reprodutivo de rotina em uma propriedade rural, uma novilha da raça Jersey, recém-adquirida, com histórico de ter sido submetida à IATF, foi avaliada para diagnóstico de gestação. No exame a novilha não estava gestante e apresentava um cisto folicular ovariano. A novilha foi submetida novamente à IATF em que foi inserido e removido um implante, com isso o cisto regrediu e ao inseminar a novilha não foi possível passar a cérvix com o aplicador. Vinte dias após a inseminação a novilha manifestou estro e ao ser avaliada para nova inseminação, foi identificado um implante solto na cavidade abdominal da novilha. Foi realizada uma laparotomia pelo flanco esquerdo e o implante foi removido. Após a recuperação da cirurgia a novilha foi inseminada e novamente e ficou gestante. Os protocolos de IATF devem ser realizados por profissionais capacitados para evitar problemas como a presente situação em que um implante foi encontrado na cavidade abdominal de uma novilha.(AU)

The fixed-time artificial insemination (FTAI) is a biotechnique with numerous advantages for cattle production. In order to perform the FTAI, the cows must receive a pharmacological protocol to induce ovulation in a synchronized way. This protocol begins with the insertion of a device impregnated with progesterone that must remain in the vaginal fundus of the cow for a specific period of time. This report aims at describing the removal of a progesterone impregnated implant that was found in the abdominal cavity of a heifer. During a routine reproductive service in a rural property, a recently acquired Jersey heifer, with history of having been submitted to FTAI, was evaluated for pregnancy diagnosis. At the exam, it was observed that the heifer was not pregnant and presented an ovarian follicular cyst. The heifer was then submitted to FTAI again, where an implant was inserted and removed. This made the cyst to recede, and upon subsequent insemination, the applicator was not able to pass the cervix. Twenty days after insemination, the heifer was in estrus and when it was evaluated for insemination, a loose implant was identified in the abdominal cavity of the heifer. A laparotomy was performed through the left flank and the implant was removed. After recovery from the surgery, the heifer was inseminated again and became pregnant. The FTAI protocols should only be carried out by trained professionals to avoid problems such as the one presented herein, where an implant was found in the abdominal cavity of a heifer.(AU)

La inseminación artificial a tiempo fijo (FTAI) es una biotécnica con numerosas ventajas para la producción de ganado. Para realizar FTAI, las hembras bovinas deben recibir un protocolo farmacológico que induzca la ovulación de forma sincronizada. Este protocolo comienza con la inserción de un dispositivo impregnado con progesterona que debe permanecer en el fondo vaginal de la hembra durante un tiempo específico. Este informe tiene como objetivo describir la extracción de un implante impregnado de progesterona que se encontró en la cavidad abdominal de una novilla. Durante la atención reproductiva de rutina en una propiedad rural, se evaluó el diagnóstico de preñez de una novilla de Jersey recién adquirida con antecedentes de someterse a FTAI. En el examen, la novilla no estaba embarazada y tenía un quiste folicular ovárico. La novilla fue nuevamente sometida a FTAI, en la cual se insertó y removió un implante, con esto el quiste retrocedió y al inseminar a la novilla no fue posible pasar el cérvix con el aplicador. Veinte días después de la inseminación, la vaquilla manifestó estro y cuando se evaluó para una nueva inseminación, se identificó un implante suelto en la cavidad abdominal de la vaquilla. Se realizó una laparotomía a través del flanco izquierdo y se extrajo el implante. Después de recuperarse de la cirugía, la novilla fue inseminada y nuevamente quedó embarazada. Los protocolos FTAI deben ser realizados por profesionales capacitados para evitar problemas como la situación actual donde se encontró un implante en la cavidad abdominal de una novilla.(AU)

Relación genética entre toros de inseminación comercializados en Chile por distintas empresas proveedoras de semen: un estudio de caso en las razas Holstein y Angus / Genetic relationship between insemination bulls marketed in Chile by two semen suppliers: a case study in Holstein and Angus breeds / Relação genética entre touros de inseminação comercializados no Chile por diferentes empresas fornecedoras de sêmen: um estudo de caso nas raças Holstein e Angus

Resumen La inseminación artificial (IA) es actualmente el manejo reproductivo preferido para la mayoría de los ganaderos que trabajan con razas deganado internacionales como Holstein Friesian y Aberdeen Angus. En relación con esto, varios autores han descrito que a lo largo de los años ha habido una alteración en parámetros de productivos, lo que se asociaría significativamente con un aumento en el grado de endogamia en los animales. Esta situación tendría su origen a nivel de los núcleos genéticos, en los que se llevan a cabo programas de mejora, debido al pequeño número de líneas familiares a las que pertenecen los animales seleccionados como reproductores. Este estudio evaluó el grado de parentesco entre toros de inseminación Holstein Friesian y Aberdeen Angus comercializados en Chile por dos empresas de distribución de semen. Se estudiaron un total de 86 toros disponibles en catálogos hasta el año 2017. La información genealógica de cada toro se remonta hasta trastatarabuelo y luego se construyó un registro genealógico consolidado para cada raza. Este análisis consideró la estimación del porcentaje de endogamia (F) de cada toro y la construcción de una matriz de parentesco aditivo (A) entre los animales. Los resultados indicaron que el 99% de los toros Holstein y el 79% de los toros Angus están emparentados. Como resultado de lo anterior, se concluye que es esencial que en Chile se inicie un programa de gestión de información genealógica para controlar adecuadamente los niveles de endogamia y poder mitigar los problemas que puedan surgir a causa de este fenómeno.

Abstract Artificial insemination (AI) is currently the preferred reproductive management for most cattle farmers working with international cattle breeds likes Holstein Friesian and Aberdeen Angus. In relation to this, several authors have described that over the years there has been an alteration in the production parameters, which would be significantly associated with an increase in the inbreeding degree of the animals. This fact would have its origin at the genetic nucleus level, in which improvement programs are carried out, due to the small number of family lines which the animals selected as breeding stock belong to. This study evaluated the degree of kinship between Holstein Friesian and Aberdeen Angus insemination bulls marketed in Chile by two semen distribution companies. A total of 86 bulls available in catalogues until 2017 were studied. The genealogical information of each bull was traced back up to great-great-grandfather and then a consolidated genealogical record was built for each breed. This analysis considered the estimation of each bull's inbreeding percentage (F) and the construction of a co-ancestry matrix among animals. Results indicated that 99% of Holstein bulls and 79% of Angus bulls, are related to some extent. As a result of the above, it is concluded that is essential that in Chile a program management of genealogical information be initiated to adequately control inbreeding problems that may arise from the use of related animals.

Resumo A inseminação artificial (IA) atualmente é o manejo reprodutivo preferido da maioria dos criadores de gado que trabalham com raças de gado internacionais, como o Holstein Friesian e o Aberdeen Angus. Nesse âmbito, vários autores assinalam que, ao longo dos anos, tem ocorrido uma alteração dos parâmetros produtivos, que poderia estar associada de forma significativa com um aumento no nível de endogamia nos animais. A origem dessa situação poderia estar nos núcleos genéticos, pois nos programas de melhoria, os animais que são selecionados como reprodutores pertencem a um reduzido número de linhas familiares. Esta pesquisa avalia os graus de parentesco entre touros para inseminação Holstein Friesian e Aberdeen Angus comercializados no Chile por duas empresas de distribuição de sêmen. No total, 86 touros foram analisados, disponíveis em catálogos de até 2017. As informações genealógicas de cada touro remontam aos tetravós. Posteriormente, foi elaborado um registro genealógico consolidado para cada raça. A análise considerou a estimação da porcentagem de endogamia (F) de cada touro e a construção de uma matriz de parentesco aditivo (A) entre os animais. Os resultados indicam que 99% dos touros Holstein e 79% dos touros Angus são aparentados. Como resultado disso, chega-se à conclusão de que é essencial se ter um programa de gestão de informações genealógicas no Chile para controlar adequadamente os níveis de endogamia e assim mitigar os possíveis problemas gerados por esse fenômeno.

Identificación por catálogo y detección molecular de bovinos Holstein portadores de braquiespina en Uruguay / Identification through sire catalogues analysis and molecular detection of brachyspina carriers in Uruguayan Holstein

Resumen El síndrome de braquiespina es una condición genética de la raza Holstein, detectada en el año 2006. Es causado por una deleción de 3.3 Kb en el gen FANCI localizado en el cromosoma bovino 21. La mutación fue identificada en poblaciones de Holstein de Europa, América del Norte y Asia. Dada la importancia económica del defecto y su amplia distribución mundial, el objetivo de este trabajo ha sido la identificación de animales portadores en el núcleo de selección genética de la raza en Uruguay y el diagnóstico molecular del alelo deletéreo en animales del rodeo nacional. En el presente estudio se analizaron 2598 registros de toros Holstein del catálogo de padres del sistema de evaluación genética lechera, los registros de toros pertenecientes a los catálogos de semen Holstein disponible para Uruguay de los años 2014 al 2018; y 71 vacas pertenecientes al rodeo general. Se encontraron 28 toros portadores de braquiespina de un total de 377 toros con información genética del catálogo de padres y cuatro vacas portadoras de un total de 71 genotipificadas en nuestro laboratorio. Se demostró una disminución en el ingreso de semen de animales portadores al país entre los años 2014 y 2018. La frecuencia significativa de animales portadores en Uruguay evidencia la necesidad de implementar estrategias que permitan eliminar gradualmente el defecto de la población.

Abstract Brachyspina syndrome is a hereditary recessive disease of recent identification in the Holstein breed. It is caused by a deletion of 3.3Kb in the FANCI gene located in the bovine chromosome 21. The mutation was identified in Holstein populations of Europe, North America and Asia. Given the economic importance of the defect and its wide distribution, the objective of this work was the identification of carrier animals in the genetic selection nucleus of the breed in Uruguay and the molecular verification of the deleterious allele in animals of the national herd. In the present study, 2598 records of Holstein bulls were analyzed from the list of parents of the dairy genetic evaluation system, records of bulls belonging to the Holstein semen catalogs available for Uruguay from 2014 to 2018; and 71 cows belonging to the general herd. Twenty-eight brachyspina carrier bulls were found of a total of 377 bulls with genetic information from the list of parents and four carrier cows of a total of 71 genotyped in our laboratory. A decrease in the income of semen from carrier animals to the country between 2014 and 2018 was demonstrated. The significant frequency of carrier animals in Uruguay evidences the need to implement strategies to gradually eliminate the population defect.

Different times to perform timed artificial insemination when using a P4/E2/eCG-based protocol in buffalo / Momentos diferentes para executar a inseminação artificial em tempo fixo utilizando o protocolo à base de P4/E2/eCG em búfalas

ABSTRACT: The aim of this study was to evaluate different times for timed artificial insemination (TAI) in buffalo submitted to a P4/E2/eCG-based protocol. In this study, 204 buffaloes were distributed into one of two groups (TAI56, n=103 and TAI64, n=101). At a random stage of the oestrous cycle (Day 0 = D0), in the morning (TAI56, a.m.) or afternoon (TAI64, p.m.), buffaloes received an intravaginal progesterone device (P4; 1.0 g) plus EB (2.0 mg i.m.). On D9 a.m. (TAI56) or p.m. (TAI64), the P4 was removed and buffaloes received PGF2a (0.53 mg i.m. sodium cloprostenol) and eCG (400 IU i.m.). On D10 a.m. (TAI56) or p.m. (TAI64), 24 h after P4 removal, buffaloes were treated with EB (1.0 mg i.m.). Buffaloes from TAI56 and TAI64 were inseminated 56 and 64 h after P4 removal (D11, p.m. and D12, a.m., respectively). Ultrasound examinations were performed on D0 to ascertain ovarian follicular status, at TAI to measure the diameter of the dominant follicle (DF) and D42 for pregnancy diagnosis. The statistical analysis was performed using the GLIMMIX procedure of SAS®. There was no difference between TAI56 and TAI64 for the diameter of the DF at TAI and the pregnancy per TAI. It was concluded that TAI 56 or 64 h after P4 removal did not affect fertility in buffaloes submitted to the induction of ovulation with EB. The present research supports that is possible to perform TAI at any time throughout the day in buffalo synchronized during the non-breeding season.

RESUMO: O objetivo deste estudo foi avaliar diferentes momentos para a realização da IATF em búfalas submetidas a um protocolo à base de P4/E2/eCG. Neste estudo, 204 búfalas foram distribuídas em um de dois grupos (IATF56, n=103 e IATF64, n=101). Em estágio aleatório do ciclo estral (Dia 0 = D0), pela manhã (IATF56, manhã) ou pela tarde (IATF64, tarde), as búfalas receberam um dispositivo intravaginal de progesterona (P4; 1,0 g) e BE (2,0 mg i.m.). No D9 pela manhã (IATF56) ou pela tarde (IATF64), a P4 foi removida e as búfalas receberam PGF2a (0,53 mg i.m. cloprostenol sódico) e eCG (400 UI i.m.). No D10 pela manhã (IATF56) ou pela tarde (IATF64), 24 h após a remoção da P4, as búfalas foram tratadas com BE (1,0 mg i.m.). As búfalas dos grupos IATF56 e IATF64 foram inseminadas 56 e 64 h após a remoção da P4 (D11, pela tarde e D12, pela manhã, respectivamente). Avaliações ultrassonográficas foram realizadas no D0 para verificar o status folicular ovariano, na IATF para medir o diâmetro do folículo dominante (FD) e no D42 para o diagnóstico de gestação. A análise estatística foi realizada utilizando o procedimento GLIMMIX do SAS®. Não houve diferença entre os grupos IATF56 e IATF64 no diâmetro do FD na IATF e na prenhez por IATF. Conclui-se que a IATF 56 ou 64 h após a remoção da P4 não afeta a fertilidade de búfalas submetidas à indução da ovulação com BE. A presente pesquisa evidencia que é possível realizar a IATF durante todo o dia em búfalas sincronizadas durante a estação reprodutiva desfavorável.

Diferentes momentos de aplicação de gonadotrofina coriônica equina em protocolo de inseminação artificial em tempo fixo para vacas de leite / Different times of application of equine chorionic gonadotropin in fixed-time artificial insemination protocols for dairy cows

Avaliaram-se diferentes momentos de aplicação da gonadotrofina coriônica equina (eCG) em protocolos de inseminação artificial em tempo fixo (IATF) para vacas de leite. Foram utilizadas 76 fêmeas, as quais receberam, no dia zero (D0) do protocolo, dispositivos intravaginais de progesterona, sendo esses retirados no D9, e os animais foram, então, distribuídos aleatoriamente em três tratamentos: T1 - aplicação de eCG no momento da retirada dos dispositivos; T2 e T3 - aplicação de eCG 48h e 24h antes da retirada dos dispositivos, respectivamente. No D10 os animais receberam 1mg de GnRH, e a IATF foi realizada 52 horas após a retirada do implante. Não houve diferença (P>0,05) para intervalo entre a retirada do implante à ovulação (72,56±3,92h), o diâmetro do maior folículo no D9 (10,88±1,49mm), o diâmetro do folículo ovulatório (15,15±1,16mm) e do segundo maior folículo (7,49±0,52mm), a taxa de crescimento folicular (1,38±0,04mm/dia), a taxa de ovulação (96,67%), o intervalo entre diâmetro final e inicial do folículo dominante (73,49±3,84h), a área de corpo lúteo (2,27±0,43cm²), a porcentagem de CL no ovário direito (53,00%) e no esquerdo (26,33%) e a taxa de gestação (33,33%). O momento da aplicação da eCG não influenciou na eficiência do protocolo. Recomenda-se a utilização da eCG no momento da retirada do implante por otimização do manejo.(AU)

Different times of application of equine chorionic gonadotropin (eCG) in Fixed-Time Artificial Insemination (FTAI) protocols for dairy cows were evaluated. A total of 76 females were used, which received intravaginal progesterone devices on day zero (D0) of the protocol, which were withdrawn on D9, and the animals were then randomly distributed in three treatments: T1- application of eCG at the time of device withdrawal; T2 and T3 - application of eCG 48h and 24h before withdrawal of the devices, respectively. On D10 the animals received 1mg of GnRH and the FTAI was carried out 52 hours after the removal of the implant. There was no difference (P>0.05) for interval from implant removal to ovulation (72.56±3.92h), diameter of the largest follicle of D9 (10.88±1.49mm), diameters of ovulatory follicle (15.15±1.16mm) and of the second largest follicle (7.49±0.52mm), follicle growth rate (1.38±0.04mm/day), ovulation rate (96.67%), interval between final diameter and initial diameter of dominant follicle (73.49±3.84h), corpus luteum area (2.27±0.43cm²), percentage of CL in right ovary (53.00%) and in left ovary (26.33%) and pregnancy rate (33.33%). The time of the application of eCG did not influence the efficiency of the protocol. We recommend the use of eCG at the time of the removal of the implant for management optimization.(AU)

Perspectives of health care professionals on artificial insemination of donor semen: appraising their knowledge and perceptions: a single institution study in Nigeria

Background: Artificial inseminations of donor semen (AID) involve use of heterologous donated semen for conception in infertile couple when indicated or in a single woman desirous of pregnancy. Its practice often requires regulation to address possible ethical and legal issues which may arise. In formulating acceptable guidelines/policies, the perspectives of health professionals and the participants should be considered. Therefore, we sought to explore the knowledge and perception of semen donation for artificial insemination among health professionals.Methods: A cross sectional study conducted on consenting health workers in a single health institution using a structured questionnaire to assess their knowledge and perception on artificial insemination of donor semen (AID). Descriptive and analytical statistics were applied to the data collected with a statistically significant value of <0.05.Results: One hundred and twenty-one health professionals completed the questionnaires. The mean age of the respondents was 27.58±5.5years. Sixty of the respondents were males while 61 were females. Eighty-four of the respondents (69.4%) demonstrated good knowledge of AID while 37 (30.6%) had poor knowledge. Cadre of health professionals and marital status influenced the knowledge of respondents. Perceptions on AID varied among the respondents mostly influenced by psycho-social factors and possible legal disputes on third party reproductive process. None of the male respondents has ever donated semen and willingness to donate semen was low; with anonymity preferred by the willing donors.Conclusions: Substantial knowledge gap of AIDS existed among health professionals which were influenced by cadre and marital status. Psycho-social factors and possible legal disputes influenced their perceptions of AID.

Assisted Reproductive Technologies in Latin America and Europe: a Comparative Analysis of Reported Databases for 2013 / Técnicas de reprodução assistida na América Latina e Europa: Uma análise comparativa das bases de dados de 2013

Abstract Objective To compare the Latin American and European assisted reproductive technology (ART) registries regarding data accessibility and quality, treatment utilization, effectiveness, safety, and quality of services. Methods We performed an ecological study using data from scientific publications of Latin American and European registries that report cycles initiated during 2013 (the most recent registries available until December of 2017). The summarized data are presented as frequencies, percentages, minimum-maximum values, and absolute numbers. Results Reporting clinics and cycle treatments were unevenly distributed between the participating countries for both registries, although access to ART is 15 times greater in Europe. In Latin America, individual services participate voluntarily reporting started cycles until cancellation, birth or miscarriage, while in Europe it varied among countries. It makes the data available from Latin America more uniform, although lesser representative when compared with European ones, given that reporting is compulsory formost countries. The cumulative live birth rate was better in Latin America. Female age, use of intracytoplasmic sperm injection (ICSI), cycles with transfer of ≥ 3 embryos, as well as multiple pregnancy rates were greater in the Latin American Register of Assisted Reproduction (RLA, in the Portuguese acronym). Assisted reproductive technology complications, such as ovarian hyperstimulation syndrome, hemorrhage, and infections were also higher in LatinAmerica, although they are extremely uncommon in both regions. Conclusion Both regions have points to improve in the quality of their reports. Latin America has produced a more uniform reporting, their clinical results are generally

Resumo Objetivo Comparar os registros das técnicas de reprodução assistida da América Latina e da Europa em relação à qualidade dos dados e acesso, utilização do tratamento, efetividade, segurança e qualidade dos serviços. Métodos Estudo ecológico usando os dados das publicações científicas dos registros da América Latina e da Europa com os ciclos iniciados durante o ano de 2013 (que são os dadosmais recentes disponíveis até dezembro de 2017). Os dados são apresentados como porcentagens, valores mínimos e máximos e números absolutos. Resultados Em comum, vemos que o número de clínicas e de ciclos de tratamento varia bastante entre os países dentro de cada região emambos os registros, embora o acesso às técnicas de reprodução assistida seja 15 vezes maior na Europa. Na América Latina, os serviços reportam voluntariamente os ciclos iniciados até o cancelamento, o nascimento ou aborto, enquanto que na Europa o que é reportado varia entre os países. Isso faz o registro da América Latina mais uniforme, apesar de ser menos representativo quando comparado ao Europeu, dado o caráter compulsório na maioria dos países deste último. A taxa de gravidez cumulativa, idade feminina, uso de injeção intracitoplasmática de espermatozoides (ICSI), ciclos comtransferência ≥ 3 embriões e taxa de gestação múltipla foram mais elevados na América Latina. Complicações da reprodução assistida, como síndrome de hiperestimulação ovariana, hemorragia, e infecções também foram mais comuns na América Latina, embora sejam incomuns em ambas as regiões. Conclusão Ambas as regiões têm pontos a melhorar na qualidade dos registros. A América Latina tem produzido um registro mais uniforme, e seus resultados clínicos são comparáveis e, algumas vezes, superiores aos Europeus. Por outro lado, a segurança do tratamento foi maior na Europa, com menores taxas de complicações, especialmente gestações múltiplas.

Clinical and blood gas analysis of calves conceived by artificial insemination, in vitro fertilization and animal cloning / Análise clínica e hemogasométrica de bezerros concebidos por inseminação artificial, fertilização in vitro e clonagem animal

In order for successful extra-uterine adaptation to occur, it is necessary for the neonate to be able to establish its respiratory functions effectively, guaranteeing efficient oxygenation and good vitality. Respiratory disorders are the major cause of death during the neonatal period in cattle, and this mortality is even more significant when it comes to calves originated by in vitro fertilization (FIV) or animal cloning (CA). Blood gas analysis assesses acid-base balance changes effectively, and when associated with the neonate's clinical examination, provides subsidies for accurate diagnosis and early treatment of neonatal maladaptation. The objective of this study was to study neonates born from artificial insemination (IA) and to compare them to calves conceived by FIV and CA, regarding blood gas and clinical examination. For that, 20 AI calves, 15 FIV calves, and 15 cloned calves were evaluated immediately after calving and at 6, 12, 24 and 48 hours of life. At all experimental times, venous blood samples were collected for blood gas and clinical examination was performed. In the postpartum evaluation, Apgar score and column length and respiratory amplitude measurements were used. IVF animals showed no alterations, resembling Group IA calves. The calves from CA showed more pronounced acidosis postpartum than expected physiological acidosis mixed for neonates, with decreasing values of bicarbonate (HCO3-), and base excess (BE) and the increase in carbon dioxide pressure (PCO2) when compared to the other groups. This disorder may have reflected lower mean values of Apgar scores and increased heart and respiratory rates. Intensive follow-up of these neonates is suggested, with monitoring by clinical and hemogasometric examination for early diagnosis of this condition and treatment based on oxygen therapy and bicarbonate replacement.(AU)

Para que ocorra adaptação extra-uterina bem sucedida é necessário que o neonato consiga estabelecer suas funções respiratórias de maneira eficaz, garantindo oxigenação eficiente e boa vitalidade. Distúrbios respiratórios são os maiores causadores de óbito durante o período neonatal em bovinos, e essa mortalidade é ainda mais expressiva quando se trata de bezerros originados por fertilização in vitro (FIV) ou clonagem animal (CA). A hemogasometria avalia alterações do equilíbrio ácido-básico de forma eficaz, e quando associada ao exame clínico do neonato, fornece subsídios para diagnóstico acurado e tratamento precoce da má adaptação neonatal. O objetivo deste trabalho foi estudar recém-nascidos bovinos originados por inseminação artificial (IA) e compará-los a bezerros concebidos por FIV e CA, no que se refere a hemogasometria e exame clínico. Para isso, foram utilizados 20 bezerros IA, 15 bezerros FIV e 15 bezerros clonados que foram avaliados imediatamente após o parto e com 6, 12, 24 e 48 horas de vida. Em todos os momentos experimentais foram colhidas amostras de sangue venoso para hemogasometria e foi realizado o exame clínico. Na avaliação pós-parto foram utilizados escore Apgar e mensurações de comprimento de coluna e amplitude respiratória. Os animais FIV não demonstraram alterações, assemelhando-se aos bezerros do Grupo IA. Os bezerros provenientes de CA apresentaram acidose pós-parto mais acentuada do que a acidose mista fisiológica esperada para neonatos, evidenciada pela diminuição dos valores de bicarbonato (HCO3-) e excesso de bases (EB) e pelo aumento de pressão parcial de dióxido de carbono (PCO2) quando comparados aos demais grupos. Esse distúrbio pode ter refletido em valores médios menores de escore Apgar e no aumento das frequências cardíaca e respiratória. Sugere-se acompanhamento intensivo desses neonatos, com monitoramento por meio do exame clínico e hemogasométrico para diagnóstico precoce dessa condição e tratamento baseado em oxigenioterapia e reposição de bicarbonato.(AU)

Ultrasound guidance versus the blind method for intrauterine catheter insemination: A randomized controlled trial / 대한생식의학회지

OBJECTIVE: The primary objective of this study was to compare clinical pregnancy rates in intrauterine insemination (IUI) treatment cycles with transabdominal ultrasound guidance during intrauterine catheter insemination (US-IUI) versus the “blind method” IUI without ultrasound guidance (BM-IUI). The secondary objective was to compare whether US-IUI had better patient tolerability and whether US-IUI made the insemination procedure easier for the clinician to perform compared to BM-IUI. METHODS: This was a randomized controlled trial done at the Reproductive Medicine Unit of General Hospital Kuala Lumpur, Malaysia. We included women aged between 25 and 40 years who underwent an IUI treatment cycle with follicle-stimulating hormone injections for controlled ovarian stimulation. RESULTS: A total of 130 patients were recruited for our study. The US-IUI group had 70 patients and the BM-IUI group had 60 patients. The clinical pregnancy rate was 10% in both groups (p> 0.995) and there were no significant difference between the groups for patient tolerability assessed by scores on a pain visual analog scale (p= 0.175) or level of difficulty for the clinician (p> 0.995). The multivariate analysis further showed no significant increase in the clinical pregnancy rate (adjusted odds ratio, 1.07; 95% confidence interval, 0.85–1.34; p= 0.558) in the US-IUI group compared to the BM-IUI group even after adjusting for potential covariates. CONCLUSION: The conventional blind method for intrauterine catheter insemination is recommended for patients undergoing IUI treatment. The use of ultrasound during the insemination procedure increased the need for trained personnel to perform ultrasonography and increased the cost, but added no extra benefits for patients or clinicians.

Comparative analysis of medical, legal, and ethical considerations for establishing a standard operating protocol for artificial insemination by donor

Artificial insemination by donor is an important means of improving the likelihood of pregnancy in couples affected by male factor infertility, but it poses medical, legal, and ethical issues due to the involvement of third parties, such as the sperm donor. In Korea, the Bioethics and Safety Act was enacted for the purpose of preventing and eliminating unethical research on germ cells, and such research was limited to matters related to the use of assisted reproductive technologies, centering on embryos, oocytes, and protecting the health of oocyte donors. However, this law is incomplete in terms of specific standards or regulations relating to the donation and receipt of sperm. In Korea, artificial insemination by donor has been carried out without a standard operating protocol for donation and receipt of sperm, which would include testing sperm donors for diseases, limiting the number of donor offspring, compensation for donations, and the role of anonymity and non-anonymity. The diversity of policies worldwide shows that each country has its unique set of guidelines tailored for its own specific needs and practical considerations. Herein, I present a standard operating protocol of medical, legal, and ethical principles for artificial insemination by donor that is suitable for domestic circumstances, along with a comparison of recommendations and guidelines of other countries concerning sperm donation issues.

Reasons of shortage for donated sperm and plan to resolve deficiencies in the sperm donor system

Sperm banking from recruited donors is an important way to help infertile couples who want to achieve pregnancy by therapeutic artificial insemination, even in the era of in vitro fertilization. Korea has the lowest population density of newborns among all Organization for Economic Co-operation and Development countries, with a total fertility rate of 1.17 and an annual birth rate of 300,000 or less as of 2017. This situation emerged due to various causes, including late marriage, fecundity problems, and air pollution. Nevertheless, we have imposed strict limits on basic research and clinical activities, including the operation of sperm banks, in the field of human reproduction under the 2005 Bioethics and Safety Act. Therefore, cryopreserved sperm for artificial insemination is almost absent in the sperm banks of major human reproduction centers. To resolve this difficult situation, as well as to increase the operation of sperm banks in Korea to the global standard level, the author evaluates the underlying reasons for donor sperm use by azoospermic patients from medical, ethical, legal, religious, and cultural perspectives, and then proposes a plan to resolve this situation including highly regulated standard operating procedures to ensure comprehensive safety practices for voluntary sperm donors and infertile couples, as well as to build an ideal sperm donation program.

Alginate encapsulation preserves the quality and fertilizing ability of Mediterranean Italian water buffalo (Bubalus bubalis) and Holstein Friesian (Bos taurus) spermatozoa after cryopreservation

The use of artificial insemination (AI) in buffalo (Bubalus bubalis) is limited by poor ovarian activity during the hot season, seasonal qualitative patterns in semen, low resistance of sperm cells in the female tract, difficulties in estrus detection, and variable estrus duration. Although AI procedures are commonly used in bovine, use of AI has been limited in buffalo. In the zootechnical field, different studies have been conducted to develop techniques for improvement of fertilizing ability of buffalo spermatozoa after AI. In this study, for the first time, the use of alginate encapsulation and cryopreservation of buffalo spermatozoa is described, and the same procedure was performed with Holstein Friesian (Bos taurus) semen. Results obtained from in vitro analyses indicate that the encapsulation process does not have detrimental effects (compared to controls) on quality parameters (membrane integrity, progressive motility, path average velocity) in either species. Similarly, there were no detrimental effects after cryopreservation in either species. The fertilizing potential of encapsulated and cryopreserved semen was evaluated after AI in 25 buffalo and 113 bovine females. Pregnancy rates were not affected in either species. The results of this study show proof of concept for the use of frozen semen controlled-release devices in buffalo.

Inquiry Survey of Infertile Couples for Sperm Donation and Artificial Insemination by Donor

BACKGROUND: Artificial insemination by donor (AID) is important to chance the pregnancy in male infertile couples by requiring appropriate medical, legal and ethical reviews. The purpose of this study was to investigate the perception changes of AID to collect the basic information for establishing the sperm donation, cryopreservation and artificial insemination management system. METHODS: We evaluated the people's thought of sperm donation, cryopreservation, and artificial insemination by donor by means of national inquiry survey from 247 infertile couples for 4 weeks from October 2016. The questionnaires were composed of 4 areas, and each questionnaire consisted of 15 items. Changes in the perception of AID by year were investigated after review of domestic journals. RESULTS: 67.6% of respondents knew the growth of couples with difficulty getting pregnant due to male infertility. 82.2% of respondents replied the necessary of sperm bank as a treatment option for infertility and 40.5% knew the donation and receipt system of sperm. In the survey of change perception of AID according to year, 37.5% and 39.5% of respondents were got information about AID from doctor in 2003 and 2004, but 49.6% from broadcasting in 2016. As a child grows up, 4.0% (2003), 9.0% (2014), and 42.8% (2016) of respondents answered to tell the child about AID. CONCLUSIONS: Infertile couple's thought of AID about the extension of opportunities for male infertility treatment is changed. In conclusion, it is necessary to establish institutional system of sperm donation, cryopreservation and artificial insemination prior to public sperm bank operation.

Testicular cytology indicates differences in Sertoli cell counts between “good freezer” and “poor freezer” bulls.

In artificial insemination, poor quality of semen unsuitable for cryopreservation and susceptibility of spermatozoa to cryodamage in crossbred bulls have been a matter of concern. Present study was designed to identify the testicular cytology indices that might be used to predict the semen quality and cryotolerance of spermatozoa in bulls. Based on the ejaculate rejection rate and sperm cryotolerance, bulls (Holstein Friesian X Tharparkar crossbred) were classified into either good (producing good quality semen with spermatozoa having good cryotolerance; n=4) or poor (producing poor quality semen with spermatozoa having poor cryotolerance; n=4). Testicular cytology was studied in all the 8 bulls using fine needle aspiration technique. Testicular cytology of good bulls and poor bulls differed significantly. The proportion of Sertoli cells was significantly higher in good bulls (25.3±1.6) compared to poor bulls (11.0±0.8). The Sertoli cell index was 46.1±5.0 in good bulls while it was only 13.8±1.3 in poor bulls. The cut off values, as determined using Receiver Operating Characteristics analysis, indicate that the bulls having testicular cytogram comprising of <15.5% Sertoli cells, <24.3 Sertoli cell index and >4.0 spermatogenic cells to Sertoli cell ratio might be a poor bull in terms of semen quality and cryotolerance of spermatozoa. The proportion of Sertoli cells in the testicular cytology had positive (P <0.05) relationship with semen quality and cryotolerance of spermatozoa.

Infertility Treatment and Congenital Malformations

Over the past 30 years, fertility treatments including fertility enhancing drugs, artificial insemination and in vitro fertilization (IVF) have been revolutionized. Thus, worldwide, more than 5 million children have been born after assisted reproductive technology (ART) and domestically, it was announced that 2.83% of all births in 2011 were associated with financial support from the National Supporting Program for the Subfertile (NSPS) administered by the Korean Ministry of Health and Welfare (KMHW). As more women seek for fertility treatment, there is growing concern about the safety of the methods. In this review, we aim to summarize the literature on the association between fertility treatment and congenital malformations with respect to subfertility, fertility enhancing medications, and different ART methods. To sum up, although ARTs are associated with slight increase in some malformations, it appears that the increase in congenital malformations is due less to ART methods per se than to the biological perturbations that generated the infertility necessitated ART to achieve pregnancy. And no particular organ system seems disproportionately affected. It is often instructive to remind all couples attempting pregnancy that the baseline malformation rate is 2~3%, compared with 3~4% in ART.

The current status of public sperm bank in Korea

Sperm banking is an important option to maintain the male fertilization capacity or induce pregnancy even though under the era of in vitro fertilization. The medical indications for sperm banking are generally consisted of 3 categories. There are cases on planning the permanent contraception like vasectomy or cancer patients to be scheduled the chemotherapy or radiotherapy as first category, male infertile patients with severe oligozoospermia or artificially harvested sperm ie, from microscopic epididymal sperm aspiration or testicular sperm extraction et cetera for the artificial insemination with husband sperm as second category, and the therapeutic donor insemination as third category. Of these three categories, the sperm donation program accompanies various complicated practical, ethical and legal issues. Therefore, highly regulated statements are mandatory in order to secure safety and the complete practices for voluntary sperm donors and infertile couples both. In aspect of administrative structure of sperm bank, there are 3 types that are national based to be established in the most of European countries and China, public based in Japan, and commercially available in the USA. Additionally, each country has different standard guidelines, regulation statements, act and law to control the sperm donation program as well as different cultural or religious background. Nevertheless, we need a consensus document to operate the sperm bank with the standard guidelines to be well revised according to each country's ethical perspectives as well as contemporary scientific evolution. This article will provide what is the Korean model for ideal sperm bank with the history of sperm cryopreservation and banking, background and prerequisite for the public sperm bank operation, and also expected effects.

Impact of Equine Chorionic Gonadotropin Associated with Temporary Weaning, Estradiol Benzoate, or Estradiol Cypionate on Timed Artificial Insemination in Primiparous Bos Indicus Cows

The study aimed to determine the impact of equine chorionic gonadotropin (eCG) associated with different timed artificial insemination (TAI) protocols on the pregnancy rate (PR) in Bos indicus cows previously treated with progesterone. Five hundred and fifty-seven primiparous cows were subjected to the following treatments: on day 0 (d0), GeCGTW (group equine Chorionic Gonadotropin+Temporary Weaning;n=178) received 0,558 g intravaginal progesterone (P4)+1.0 mg of estradiol benzoate (EB) (IM); on d8 (P4 removal+0,075 mg D-cloprostenol + 400 IU eCG + TW for 48 h); on d10, TAI + calves return to dam; GeCGEB (group equine Chorionic Gonadotropin+Estradiol benzoate; n=176) the same as GeCGTW without TW + application of 1.0 mg of EB on d9; GeCGEC (group equine Chorionic Gonadotropin+Estradiol Cypionate; n=203), the same as GeCGTW without TW+1.5 mg EC (IM). On d35, post TAI, pregnancy diagnosis (PD) was performed. Non-pregnant animals remained under clean-up bulls for 90 days. After this period, the animals were subjected to PD using ultrasound. The PR of TAI was 51.1%, 47.1%, and 47.8% for GeCGTW, GeCGEB24, and GeCGEC (P>0.05) respectively. The PR under clean-up bulls was 88.3%, 47.3%, and 31.1% (P<0.05). The final PR (TAI+clean-up bulls) of the groups was 94.4%, 72.1%, and 64.0%, respectively (P<0.05). It was concluded that no differences in PR among the protocols related to TAI were detected; PR in the GeCGTW protocol under clean-up bulls was higher compared to others (P<0.05); the overall PR of cows subjected to TAI+clean-up bulls was significantly higher in GeCGTW than in the other groups.