MSCAGR (Animal Science) / Department of Animal Science / The aims of the study were to evaluate the effects of dilution and chilled storage time on the quality of semen, and of different artificial insemination techniques on fertility in artificially inseminated indigenous Venda does. Fresh semen was collected using an artificial vagina from three Boer bucks aged 4±1.55 years once every four days during July and August 2016. Semen was pooled and samples were divided into two equal parts, which were extended using Biladyl® extender at ratios of 1:5 and 1:10 v/v (semen to extender), before refrigeration for 120 hours at 5 °C. The fresh undiluted semen and freshly extended semen were evaluated in six replicates for sperm motility, live-dead and sperm morphology using the Sperm Class Analyzer (SCA). Extended semen continued to be evaluated at 24 hour intervals for 120 hours. Ninety indigenous Venda does were obtained from different flocks in the Vhembe district and kept intensively in one 10 m x 40 m pen at the University of Venda experimental farm in the goat feedlot. The does were fed and watered ad libitum. After acclimatization for 14 days, estrus was synchronized using a controlled internal drug release (CIDR) containing 0.3 g of progesterone. Upon removal of the CIDR, does were injected 10 mg of PGF2α (Lutalyse® dinoprost tromethamine) Sterile Solution. At 24 hours after the removal of the CIDR, the does were injected intramuscularly with 300 international units (IU) of equine chorionic gonadotrophin (eCG). Forty eight hours after the removal of the progesterone, freshly collected and diluted (1:5 ratio ~150x106 sperm/ml), five day-stored semen were used to inseminate the does using cervical (CAI), trans-cervical (TAI), and laparoscopic artificial (LAI) insemination methods in a complete randomized design (CRD) with a 2 X 3 factorial arrangement of the treatments with 15 replications per treatment. The does were tested for pregnancy after 30 days using ultrasonography. Analyses of variance was performed on the pregnancy, kidding rates and on prolificacy using the GLM procedure of Minitab (Minitab 2013). Significant differences in all motility parameters were observed between the extension ratios and storage time (P<0.01). There were significant interactions between the extension ratio and storage time (P<0.05) on the sub-population of sperm cells with non-progressive motility (NON-P). Significant (P<0.01) interaction was observed between the semen extension ratio and storage time on medium and slow spermatozoa (P<0.01). The method of insemination did not (P>0.05) affect fertility, though both pregnancy and kidding rates numerically decreased in the order laparoscopic insemination (LAI)≥ trans-cervical insemination (TAI)≥ cervical insemination (CAI). Overall, 71% kidding rate was achieved.

O objetivo do presente estudo foi verificar a influência de diferentes pontos de risco de contaminação bacteriana durante a coleta do ejaculado suíno e seus efeitos sobre a qualidade da dose inseminante (DI). O experimento foi realizado em quatro centros de difusão genética (CDG), nos quais as coletas dos ejaculados foram observadas buscando possíveis pontos de risco de contaminação. Posteriormente, o sêmen in natura e 2 DIs, provenientes da coleta observada, foram avaliados no que se refere à quantificação bacteriana, morfologia e motilidade espermática e pH. Além do ejaculado, amostras de água e diluente foram avaliadas quanto ao número de unidades formadoras de colônia (UFC). Pêlos prepuciais compridos (>1,0 cm), a higiene da luva de coleta, líquido pingando pela mão do coletador para o interior do recipiente de coleta e a duração da coleta foram os 4, dentre os 12 fatores avaliados, que levaram a um aumento no percentual de ejaculados com valor superior a 220 UFC mL-1 de mesófilos aeróbios (P<0,05). Foi avaliado o efeito isolado ou associado de sete fatores (reprodutor sujo, óstio prepucial sujo, divertículo prepucial grande, pêlos prepuciais compridos, luva de coleta suja, pingos pela mão do coletador para dentro do recipiente de coleta e pênis escapou durante a coleta) que pudessem resultar diretamente na contaminação do ejaculado. Houve aumento significativo do número de ejaculados com contaminação superior a 220 UFC mL-1, a partir da associação de 2 fatores, quando comparados aos ejaculados obtidos em coletas sem nenhum fator predisponente para contaminação. Ao classificar as DIs conforme o grau de contaminação do diluente foram observadas redução na motilidade e no pH e aumento nas alterações de acrossoma das DIs, ao longo das 168 horas de armazenamento, no grupo cujo grau de contaminação do diluente foi superior a 14000 UFC mL-1 versus o grupo com contaminação inferior a 330 UFC mL-1. Doses inseminantes provenientes de ejaculados mais contaminados apresentaram maior grau de contaminação bacteriana. Aparentemente, quando o ejaculado é coletado com um protocolo de contaminação mínima, dificilmente seu grau de contaminação será capaz de produzir efeitos deletérios na qualidade da DI, exceto quando a origem da contaminação for proveniente de falhas higiênicas na linha de processamento das mesmas. A produção de DIs com alta qualidade do ponto de vista bacteriano, somente será possível com um rigoroso controle higiênico na linha de processamento, principalmente no que diz respeito à água e ao diluente, associados a um protocolo de contaminação mínima durante a coleta. / The aim of this study was to check the influence of different risk of factors for bacterial contamination during the collection of ejaculate and the effects in boar extended semen quality. The experiment was conducted in four boar studs, where semen collection was observed, searching for possible risk of factors for bacterial contamination. The ejaculate and two extended semen doses, deriving from the observed collection, were evaluated in regard to numbers of colony-forming units (CFU), sperm morphology and motility, and pH. Water and extender samples were also evaluated for CFU. Long preputial hair (>1 cm), the hygiene of the collection glove, liquid trickling from the hand of the technician into the semen container and the duration of the collection were the four, from twelve factors evaluated, that lead to an increase in the percentage of ejaculates with more than 220 CFU mL-1 of aerobic mesophiles (P<0.05). The isolated or combined effect of seven factors (bad hygiene of boars, dirty preputial ostium, large preputial diverticulum, long preputial hair, dirty collecting glove, liquid trickling from the hand of the technician into the semen container and escape penis during the collection), that could directly result in the contamination of ejaculates was evaluated. There was a significant increase in the number of ejaculates contaminated with more than 220 CFU mL-1 when two or more factors were associated, compared to ejaculates obtained from collections without any predisponent factors. When the extended semen doses were classified according to the degree of contamination of the extender, a decrease in motility and pH and an increase on acrosome alterations in extended semen, during 168 hours of storage, were verified in the group where the degree of contamination was higher than 14,000 CFU mL-1 versus the group with lower than 330 CFU mL-1. Extended semen derived from more contaminated ejaculates showed a higher degree of bacterial contamination. Apparently, when the ejaculate was collected with minimum contamination protocol, its degree of contamination will hardly be able to produce effects in the extended semen quality, unless when the source of contamination was hygienic failure in the processing. The production of semen extended with high quality in the bacterial point of view will only be possible with a strict hygienic control in the processing, mostly in respect to water and extender, associated with minimum contamination protocol during the collection.

O objetivo do presente estudo foi verificar a influência de diferentes pontos de risco de contaminação bacteriana durante a coleta do ejaculado suíno e seus efeitos sobre a qualidade da dose inseminante (DI). O experimento foi realizado em quatro centros de difusão genética (CDG), nos quais as coletas dos ejaculados foram observadas buscando possíveis pontos de risco de contaminação. Posteriormente, o sêmen in natura e 2 DIs, provenientes da coleta observada, foram avaliados no que se refere à quantificação bacteriana, morfologia e motilidade espermática e pH. Além do ejaculado, amostras de água e diluente foram avaliadas quanto ao número de unidades formadoras de colônia (UFC). Pêlos prepuciais compridos (>1,0 cm), a higiene da luva de coleta, líquido pingando pela mão do coletador para o interior do recipiente de coleta e a duração da coleta foram os 4, dentre os 12 fatores avaliados, que levaram a um aumento no percentual de ejaculados com valor superior a 220 UFC mL-1 de mesófilos aeróbios (P<0,05). Foi avaliado o efeito isolado ou associado de sete fatores (reprodutor sujo, óstio prepucial sujo, divertículo prepucial grande, pêlos prepuciais compridos, luva de coleta suja, pingos pela mão do coletador para dentro do recipiente de coleta e pênis escapou durante a coleta) que pudessem resultar diretamente na contaminação do ejaculado. Houve aumento significativo do número de ejaculados com contaminação superior a 220 UFC mL-1, a partir da associação de 2 fatores, quando comparados aos ejaculados obtidos em coletas sem nenhum fator predisponente para contaminação. Ao classificar as DIs conforme o grau de contaminação do diluente foram observadas redução na motilidade e no pH e aumento nas alterações de acrossoma das DIs, ao longo das 168 horas de armazenamento, no grupo cujo grau de contaminação do diluente foi superior a 14000 UFC mL-1 versus o grupo com contaminação inferior a 330 UFC mL-1. Doses inseminantes provenientes de ejaculados mais contaminados apresentaram maior grau de contaminação bacteriana. Aparentemente, quando o ejaculado é coletado com um protocolo de contaminação mínima, dificilmente seu grau de contaminação será capaz de produzir efeitos deletérios na qualidade da DI, exceto quando a origem da contaminação for proveniente de falhas higiênicas na linha de processamento das mesmas. A produção de DIs com alta qualidade do ponto de vista bacteriano, somente será possível com um rigoroso controle higiênico na linha de processamento, principalmente no que diz respeito à água e ao diluente, associados a um protocolo de contaminação mínima durante a coleta. / The aim of this study was to check the influence of different risk of factors for bacterial contamination during the collection of ejaculate and the effects in boar extended semen quality. The experiment was conducted in four boar studs, where semen collection was observed, searching for possible risk of factors for bacterial contamination. The ejaculate and two extended semen doses, deriving from the observed collection, were evaluated in regard to numbers of colony-forming units (CFU), sperm morphology and motility, and pH. Water and extender samples were also evaluated for CFU. Long preputial hair (>1 cm), the hygiene of the collection glove, liquid trickling from the hand of the technician into the semen container and the duration of the collection were the four, from twelve factors evaluated, that lead to an increase in the percentage of ejaculates with more than 220 CFU mL-1 of aerobic mesophiles (P<0.05). The isolated or combined effect of seven factors (bad hygiene of boars, dirty preputial ostium, large preputial diverticulum, long preputial hair, dirty collecting glove, liquid trickling from the hand of the technician into the semen container and escape penis during the collection), that could directly result in the contamination of ejaculates was evaluated. There was a significant increase in the number of ejaculates contaminated with more than 220 CFU mL-1 when two or more factors were associated, compared to ejaculates obtained from collections without any predisponent factors. When the extended semen doses were classified according to the degree of contamination of the extender, a decrease in motility and pH and an increase on acrosome alterations in extended semen, during 168 hours of storage, were verified in the group where the degree of contamination was higher than 14,000 CFU mL-1 versus the group with lower than 330 CFU mL-1. Extended semen derived from more contaminated ejaculates showed a higher degree of bacterial contamination. Apparently, when the ejaculate was collected with minimum contamination protocol, its degree of contamination will hardly be able to produce effects in the extended semen quality, unless when the source of contamination was hygienic failure in the processing. The production of semen extended with high quality in the bacterial point of view will only be possible with a strict hygienic control in the processing, mostly in respect to water and extender, associated with minimum contamination protocol during the collection.

O objetivo do presente estudo foi verificar a influência de diferentes pontos de risco de contaminação bacteriana durante a coleta do ejaculado suíno e seus efeitos sobre a qualidade da dose inseminante (DI). O experimento foi realizado em quatro centros de difusão genética (CDG), nos quais as coletas dos ejaculados foram observadas buscando possíveis pontos de risco de contaminação. Posteriormente, o sêmen in natura e 2 DIs, provenientes da coleta observada, foram avaliados no que se refere à quantificação bacteriana, morfologia e motilidade espermática e pH. Além do ejaculado, amostras de água e diluente foram avaliadas quanto ao número de unidades formadoras de colônia (UFC). Pêlos prepuciais compridos (>1,0 cm), a higiene da luva de coleta, líquido pingando pela mão do coletador para o interior do recipiente de coleta e a duração da coleta foram os 4, dentre os 12 fatores avaliados, que levaram a um aumento no percentual de ejaculados com valor superior a 220 UFC mL-1 de mesófilos aeróbios (P<0,05). Foi avaliado o efeito isolado ou associado de sete fatores (reprodutor sujo, óstio prepucial sujo, divertículo prepucial grande, pêlos prepuciais compridos, luva de coleta suja, pingos pela mão do coletador para dentro do recipiente de coleta e pênis escapou durante a coleta) que pudessem resultar diretamente na contaminação do ejaculado. Houve aumento significativo do número de ejaculados com contaminação superior a 220 UFC mL-1, a partir da associação de 2 fatores, quando comparados aos ejaculados obtidos em coletas sem nenhum fator predisponente para contaminação. Ao classificar as DIs conforme o grau de contaminação do diluente foram observadas redução na motilidade e no pH e aumento nas alterações de acrossoma das DIs, ao longo das 168 horas de armazenamento, no grupo cujo grau de contaminação do diluente foi superior a 14000 UFC mL-1 versus o grupo com contaminação inferior a 330 UFC mL-1. Doses inseminantes provenientes de ejaculados mais contaminados apresentaram maior grau de contaminação bacteriana. Aparentemente, quando o ejaculado é coletado com um protocolo de contaminação mínima, dificilmente seu grau de contaminação será capaz de produzir efeitos deletérios na qualidade da DI, exceto quando a origem da contaminação for proveniente de falhas higiênicas na linha de processamento das mesmas. A produção de DIs com alta qualidade do ponto de vista bacteriano, somente será possível com um rigoroso controle higiênico na linha de processamento, principalmente no que diz respeito à água e ao diluente, associados a um protocolo de contaminação mínima durante a coleta. / The aim of this study was to check the influence of different risk of factors for bacterial contamination during the collection of ejaculate and the effects in boar extended semen quality. The experiment was conducted in four boar studs, where semen collection was observed, searching for possible risk of factors for bacterial contamination. The ejaculate and two extended semen doses, deriving from the observed collection, were evaluated in regard to numbers of colony-forming units (CFU), sperm morphology and motility, and pH. Water and extender samples were also evaluated for CFU. Long preputial hair (>1 cm), the hygiene of the collection glove, liquid trickling from the hand of the technician into the semen container and the duration of the collection were the four, from twelve factors evaluated, that lead to an increase in the percentage of ejaculates with more than 220 CFU mL-1 of aerobic mesophiles (P<0.05). The isolated or combined effect of seven factors (bad hygiene of boars, dirty preputial ostium, large preputial diverticulum, long preputial hair, dirty collecting glove, liquid trickling from the hand of the technician into the semen container and escape penis during the collection), that could directly result in the contamination of ejaculates was evaluated. There was a significant increase in the number of ejaculates contaminated with more than 220 CFU mL-1 when two or more factors were associated, compared to ejaculates obtained from collections without any predisponent factors. When the extended semen doses were classified according to the degree of contamination of the extender, a decrease in motility and pH and an increase on acrosome alterations in extended semen, during 168 hours of storage, were verified in the group where the degree of contamination was higher than 14,000 CFU mL-1 versus the group with lower than 330 CFU mL-1. Extended semen derived from more contaminated ejaculates showed a higher degree of bacterial contamination. Apparently, when the ejaculate was collected with minimum contamination protocol, its degree of contamination will hardly be able to produce effects in the extended semen quality, unless when the source of contamination was hygienic failure in the processing. The production of semen extended with high quality in the bacterial point of view will only be possible with a strict hygienic control in the processing, mostly in respect to water and extender, associated with minimum contamination protocol during the collection.

A bacteriospermia pode prejudicar a qualidade das doses de sêmen suíno. Desta forma, a adição de antimicrobianos (ATM) aos diluentes de sêmen é imprescindível para a manutenção da qualidade das doses inseminantes. Contudo, a crescente ocorrência de resistência bacteriana tem impulsionado a redução do uso de ATM na suinocultura. Nesse sentido, o armazenamento das doses inseminantes em baixas temperaturas pode ser uma alternativa para a remoção dos ATM dos diluentes comerciais. Sendo assim, no presente estudo, foram realizados dois experimentos para avaliar a qualidade espermática e a contagem de unidades formadoras de colônias (UFC/mL) de doses de sêmen suíno submetidas a baixas temperaturas de armazenamento, na ausência ou presença de ATM. No experimento 1, as motilidades (total e progressiva) das doses com ATM foram maiores conforme aumentou a temperatura de armazenamento (P<0,01). Nas doses sem ATM, as motilidades foram inferiores nas mantidas a 5 °C do que nas demais (P<0,05). O número de UFC/mL foi menor nas doses sem ATM mantidas a 5 e 10 °C do que a 17 °C (P<0,05), mas não houve diferença entre as temperaturas de armazenamento nas doses com ATM (P>0,05). As integridades de acrossoma e de membrana plasmática não foram afetadas (P>0,05) pelo uso de ATM, mas foram influenciadas pela temperatura de armazenamento (P<0,0001) No experimento 2, os machos foram categorizados em BONS e RUINS de acordo com a motilidade progressiva das doses com ATM armazenadas a 5 °C nas 120 h, sendo investigado o efeito dessas categorias sobre as variáveis estudadas. A motilidade total das doses armazenadas a 17 °C foi superior à das mantidas a 5 °C diluídas sem ATM (P<0,05). Os percentuais de motilidade progressiva e de acrossomas normais foram superiores nas doses mantidas a 17 °C do que nas mantidas a 5 °C, com ou sem ATM (P<0,05). O número de UFC/ml foi maior nas doses diluídas sem ATM do que nas demais (P<0,05). Após a categorização dos machos, as motilidades (total e progressiva) foram maiores nos machos BONS do que nos RUINS (P<0,05), sem diferença significativa (P>0,05) nas integridades (acrossomal e de membrana plasmática). Apesar de a qualidade espermática ter sido afetada negativamente pelas baixas temperaturas, o armazenamento das doses de sêmen suíno a 5 °C é possível, uma vez que foi mantida a viabilidade espermática in vitro, por até 5 dias, acima do nível mínimo considerado adequado para a inseminação artificial. Contudo, o uso de doses sem antimicrobianos ainda precisa de otimização, posto que que as baixas temperaturas de armazenamento reduzem, mas não inibem por completo o crescimento bacteriano. / Bacteriospermia can impair boar semen dose quality. Thus, the addition of antibiotics (ATB) is indispensable for maintaining semen doses quality. Nevertheless, growing bacterial resistance occurrence have had driven to a reduction in use of ATB in pig industry. In this sense, storage of semen doses at low temperature may be an alternative to removal ATB of commercial semen extenders. Therefore, the aim of the present study was to assess sperm quality and number of colony-forming units (CFU mL-1) in boar semen doses stored at low storage temperatures with or without ATB, in two experiments. In experiment 1, in semen doses with ATB, total and progressive motility increased as the storage temperature increased (P<0.01). In semen doses without ATB, total and progressive motility were observed to be lower when stored at 5 °C than at 10 and 17 °C (P<0.05). The number of CFU mL-1 was lower in semen doses without ATB stored at 5 and 10 °C than at 17 °C (P<0.05), but there was no difference among storage temperatures in doses with ATB (P>0.05). Acrosome and sperm membrane integrity were not influenced (P>0.05) by using ATB, but they were influenced by storage temperature (P<0,0001) In experiment 2, boars were grouped in GOOD and POOR according to progressive motility in doses stored for up to 120 h at 5 °C. So, the effect of this classification on assessed variables, was investigated. Total motility was higher in doses stored at 17 °C than in doses without ATB stored at 5 °C (P<0.05). The percentages of progressive motility and normal acrosomes were higher in doses stored at 17 °C than in doses stored at 5 °C, with or without ATB (P<0.05). The number of CFU mL-1 was higher in doses without ATB than in remaining ones (P<0.05). Total and progressive motility were observed to be higher in GOOD than in POOR boars (P<0.05). There was no difference between groups of boars in acrosome and membrane integrity (P>0.05). Despite sperm quality was negatively affected by low temperatures, the storage of boar semen doses at 5 °C is possible, since sperm viability in vitro was maintained for up to 5 days, fulfilling the requirements of semen quality to be used in artificial insemination. Nevertheless, the use of semen doses without ATB will need optimization, since low storage temperatures decreased bacterial growth, but not completely inhibit it.

A bacteriospermia pode prejudicar a qualidade das doses de sêmen suíno. Desta forma, a adição de antimicrobianos (ATM) aos diluentes de sêmen é imprescindível para a manutenção da qualidade das doses inseminantes. Contudo, a crescente ocorrência de resistência bacteriana tem impulsionado a redução do uso de ATM na suinocultura. Nesse sentido, o armazenamento das doses inseminantes em baixas temperaturas pode ser uma alternativa para a remoção dos ATM dos diluentes comerciais. Sendo assim, no presente estudo, foram realizados dois experimentos para avaliar a qualidade espermática e a contagem de unidades formadoras de colônias (UFC/mL) de doses de sêmen suíno submetidas a baixas temperaturas de armazenamento, na ausência ou presença de ATM. No experimento 1, as motilidades (total e progressiva) das doses com ATM foram maiores conforme aumentou a temperatura de armazenamento (P<0,01). Nas doses sem ATM, as motilidades foram inferiores nas mantidas a 5 °C do que nas demais (P<0,05). O número de UFC/mL foi menor nas doses sem ATM mantidas a 5 e 10 °C do que a 17 °C (P<0,05), mas não houve diferença entre as temperaturas de armazenamento nas doses com ATM (P>0,05). As integridades de acrossoma e de membrana plasmática não foram afetadas (P>0,05) pelo uso de ATM, mas foram influenciadas pela temperatura de armazenamento (P<0,0001) No experimento 2, os machos foram categorizados em BONS e RUINS de acordo com a motilidade progressiva das doses com ATM armazenadas a 5 °C nas 120 h, sendo investigado o efeito dessas categorias sobre as variáveis estudadas. A motilidade total das doses armazenadas a 17 °C foi superior à das mantidas a 5 °C diluídas sem ATM (P<0,05). Os percentuais de motilidade progressiva e de acrossomas normais foram superiores nas doses mantidas a 17 °C do que nas mantidas a 5 °C, com ou sem ATM (P<0,05). O número de UFC/ml foi maior nas doses diluídas sem ATM do que nas demais (P<0,05). Após a categorização dos machos, as motilidades (total e progressiva) foram maiores nos machos BONS do que nos RUINS (P<0,05), sem diferença significativa (P>0,05) nas integridades (acrossomal e de membrana plasmática). Apesar de a qualidade espermática ter sido afetada negativamente pelas baixas temperaturas, o armazenamento das doses de sêmen suíno a 5 °C é possível, uma vez que foi mantida a viabilidade espermática in vitro, por até 5 dias, acima do nível mínimo considerado adequado para a inseminação artificial. Contudo, o uso de doses sem antimicrobianos ainda precisa de otimização, posto que que as baixas temperaturas de armazenamento reduzem, mas não inibem por completo o crescimento bacteriano. / Bacteriospermia can impair boar semen dose quality. Thus, the addition of antibiotics (ATB) is indispensable for maintaining semen doses quality. Nevertheless, growing bacterial resistance occurrence have had driven to a reduction in use of ATB in pig industry. In this sense, storage of semen doses at low temperature may be an alternative to removal ATB of commercial semen extenders. Therefore, the aim of the present study was to assess sperm quality and number of colony-forming units (CFU mL-1) in boar semen doses stored at low storage temperatures with or without ATB, in two experiments. In experiment 1, in semen doses with ATB, total and progressive motility increased as the storage temperature increased (P<0.01). In semen doses without ATB, total and progressive motility were observed to be lower when stored at 5 °C than at 10 and 17 °C (P<0.05). The number of CFU mL-1 was lower in semen doses without ATB stored at 5 and 10 °C than at 17 °C (P<0.05), but there was no difference among storage temperatures in doses with ATB (P>0.05). Acrosome and sperm membrane integrity were not influenced (P>0.05) by using ATB, but they were influenced by storage temperature (P<0,0001) In experiment 2, boars were grouped in GOOD and POOR according to progressive motility in doses stored for up to 120 h at 5 °C. So, the effect of this classification on assessed variables, was investigated. Total motility was higher in doses stored at 17 °C than in doses without ATB stored at 5 °C (P<0.05). The percentages of progressive motility and normal acrosomes were higher in doses stored at 17 °C than in doses stored at 5 °C, with or without ATB (P<0.05). The number of CFU mL-1 was higher in doses without ATB than in remaining ones (P<0.05). Total and progressive motility were observed to be higher in GOOD than in POOR boars (P<0.05). There was no difference between groups of boars in acrosome and membrane integrity (P>0.05). Despite sperm quality was negatively affected by low temperatures, the storage of boar semen doses at 5 °C is possible, since sperm viability in vitro was maintained for up to 5 days, fulfilling the requirements of semen quality to be used in artificial insemination. Nevertheless, the use of semen doses without ATB will need optimization, since low storage temperatures decreased bacterial growth, but not completely inhibit it.

A bacteriospermia pode prejudicar a qualidade das doses de sêmen suíno. Desta forma, a adição de antimicrobianos (ATM) aos diluentes de sêmen é imprescindível para a manutenção da qualidade das doses inseminantes. Contudo, a crescente ocorrência de resistência bacteriana tem impulsionado a redução do uso de ATM na suinocultura. Nesse sentido, o armazenamento das doses inseminantes em baixas temperaturas pode ser uma alternativa para a remoção dos ATM dos diluentes comerciais. Sendo assim, no presente estudo, foram realizados dois experimentos para avaliar a qualidade espermática e a contagem de unidades formadoras de colônias (UFC/mL) de doses de sêmen suíno submetidas a baixas temperaturas de armazenamento, na ausência ou presença de ATM. No experimento 1, as motilidades (total e progressiva) das doses com ATM foram maiores conforme aumentou a temperatura de armazenamento (P<0,01). Nas doses sem ATM, as motilidades foram inferiores nas mantidas a 5 °C do que nas demais (P<0,05). O número de UFC/mL foi menor nas doses sem ATM mantidas a 5 e 10 °C do que a 17 °C (P<0,05), mas não houve diferença entre as temperaturas de armazenamento nas doses com ATM (P>0,05). As integridades de acrossoma e de membrana plasmática não foram afetadas (P>0,05) pelo uso de ATM, mas foram influenciadas pela temperatura de armazenamento (P<0,0001) No experimento 2, os machos foram categorizados em BONS e RUINS de acordo com a motilidade progressiva das doses com ATM armazenadas a 5 °C nas 120 h, sendo investigado o efeito dessas categorias sobre as variáveis estudadas. A motilidade total das doses armazenadas a 17 °C foi superior à das mantidas a 5 °C diluídas sem ATM (P<0,05). Os percentuais de motilidade progressiva e de acrossomas normais foram superiores nas doses mantidas a 17 °C do que nas mantidas a 5 °C, com ou sem ATM (P<0,05). O número de UFC/ml foi maior nas doses diluídas sem ATM do que nas demais (P<0,05). Após a categorização dos machos, as motilidades (total e progressiva) foram maiores nos machos BONS do que nos RUINS (P<0,05), sem diferença significativa (P>0,05) nas integridades (acrossomal e de membrana plasmática). Apesar de a qualidade espermática ter sido afetada negativamente pelas baixas temperaturas, o armazenamento das doses de sêmen suíno a 5 °C é possível, uma vez que foi mantida a viabilidade espermática in vitro, por até 5 dias, acima do nível mínimo considerado adequado para a inseminação artificial. Contudo, o uso de doses sem antimicrobianos ainda precisa de otimização, posto que que as baixas temperaturas de armazenamento reduzem, mas não inibem por completo o crescimento bacteriano. / Bacteriospermia can impair boar semen dose quality. Thus, the addition of antibiotics (ATB) is indispensable for maintaining semen doses quality. Nevertheless, growing bacterial resistance occurrence have had driven to a reduction in use of ATB in pig industry. In this sense, storage of semen doses at low temperature may be an alternative to removal ATB of commercial semen extenders. Therefore, the aim of the present study was to assess sperm quality and number of colony-forming units (CFU mL-1) in boar semen doses stored at low storage temperatures with or without ATB, in two experiments. In experiment 1, in semen doses with ATB, total and progressive motility increased as the storage temperature increased (P<0.01). In semen doses without ATB, total and progressive motility were observed to be lower when stored at 5 °C than at 10 and 17 °C (P<0.05). The number of CFU mL-1 was lower in semen doses without ATB stored at 5 and 10 °C than at 17 °C (P<0.05), but there was no difference among storage temperatures in doses with ATB (P>0.05). Acrosome and sperm membrane integrity were not influenced (P>0.05) by using ATB, but they were influenced by storage temperature (P<0,0001) In experiment 2, boars were grouped in GOOD and POOR according to progressive motility in doses stored for up to 120 h at 5 °C. So, the effect of this classification on assessed variables, was investigated. Total motility was higher in doses stored at 17 °C than in doses without ATB stored at 5 °C (P<0.05). The percentages of progressive motility and normal acrosomes were higher in doses stored at 17 °C than in doses stored at 5 °C, with or without ATB (P<0.05). The number of CFU mL-1 was higher in doses without ATB than in remaining ones (P<0.05). Total and progressive motility were observed to be higher in GOOD than in POOR boars (P<0.05). There was no difference between groups of boars in acrosome and membrane integrity (P>0.05). Despite sperm quality was negatively affected by low temperatures, the storage of boar semen doses at 5 °C is possible, since sperm viability in vitro was maintained for up to 5 days, fulfilling the requirements of semen quality to be used in artificial insemination. Nevertheless, the use of semen doses without ATB will need optimization, since low storage temperatures decreased bacterial growth, but not completely inhibit it.