Effect of retained placenta and clinical mastitis on reproduction parameters, immune response, and steroidogenic receptors gene expression in postpartum crossbred dairy cows.

The purpose of this study was to evaluate the effect of retained placenta (RP) and clinical mastitis (CM) on the reproductive efficiency of crossbred dairy cows during the postpartum period and the effect in some innate immune system indicators. For this, two experiments were carried out. In the first, a total of 232 cows were evaluated and divided as: healthy control (n = 184), RP (n = 22), and CM (n = 26) groups. The RP and CM was evaluated until 30 days postpartum (DPP) and reproductive rates were measured. In experiment 2, cows were divided in control (n = 10), RP (n = 10), and CM (n = 30) groups. Between 40 and 50 DPP, clinical, gynecological examination and endometrial cytobrush were performed to evaluate subclinical endometritis (SE) and gene expression of interleukins 1ß (IL-1ß) and 6 (IL-6), chemokine ligand 5 (CCL5), estrogen α (ESR1), and progesterone (PGR) receptors by qRT-PCR analysis. In experiment 1, the conception rate at 1st artificial insemination (AI) was lower in RP and CM groups and pregnancy rate at 150 days decreased in CM group. Calving-to-1st AI interval and days open were shorter in healthy cows. In experiment 2, the occurrence of SE was 26.7% and higher in RP and CM groups. The expression of IL-1ß increased in RP and CM groups, while IL-6 was less expressed in RP group. The CCL5, ESR1, and PGR were similar between groups. In conclusion, cows with RP and CM had their reproductive efficiency negatively affected and had they initial pro-inflammatory response improved by the increase of IL-ß.

The effect of n- 3 polyunsaturated fatty acid supplementation on immune and reproductive parameters in dairy cows: a review / O efeito da suplementação de ácidos graxos poliinsaturados n-3 sobre os parâmetros imunológicos e reprodutivos em vacas leiteiras: revisão de literatura

In dairy cattle, supplementation with polyunsaturated fatty acids (PUFAs) is considered to be an important tool to decrease the negative energy balance of periparturient dairy cows and improve the reproductive and immune systems. The most common PUFAs added to ruminant diets are omega 3 (n-3 PUFA) as linolenic acid and omega 6 (n-6 PUFA) as linoleic acid. This paper aims to review the potential effects of n-3 PUFA. We consider the effects of n-3 PUFA on the bovine immune system, especially on immune cells, and on in vivo and in vitro reproductive parameters, emphasizing how n-3 PUFAs act as modulators through one or more molecular mechanisms. The incorporation of n-3 PUFA in the dairy cow diet has positive effects on animal fertility and immunity. Future research on n-3 PUFA should be more explored concerning reproduction and immune function, starting from the investigation of basic biology to their potential for application in the clinical and preventive medicine fields.(AU)

Em rebanhos leiteiros, a suplementação com ácidos graxos poliinsaturados (PUFAs) é considerada uma ferramenta importante para diminuir o balanço energético negativo de vacas leiteiras durante o periparto e contribuir para a reprodução e sistema imunológico. Os efeitos da suplementação com PUFA sobre estes sistemas têm sido pouco explorados na literatura. Os PUFAs mais comuns na dieta dos ruminantes são Ômega-3 (n-3 PUFA) como ácido linolênico e Ômega-6 (n-6 PUFA) como ácido linoleico. Esta revisão abordará os aspectos gerais do n-3 PUFA, seus efeitos mais relevantes no sistema imune, principalmente seus efeitos nas células imunes, bem como seus efeitos na parte reprodutiva, tanto in vivo como in vitro, enfatizando a ação do n-3 PUFA através de mecanismos moleculares. A incorporação de n-3 PUFAs na dieta de vacas leiteiras exerce efeitos positivos na fertilidade e imunidade. Mais estudos a fim de explorar a função do n-3 PUFA na modulação do sistema imune e parâmetros reprodutivos, desde a investigação da biologia básica até a aplicação a campo de modo clínico e preventivo, devem ser requeridos.(AU)

Systemic changes caused by artificial insemination in beef cows (Bos indicus) and their impact on animal welfare / Alterações sistêmicas causadas pela inseminação artificial em vacas de corte (Bos indicus) e seu impacto no bem-estar animal

This study aimed to verify if the process of artificial insemination (AI) characterized here as animal immobilization, the passage of the semen applicator through the cervix, and deposition of the semen in the uterus, affected cows' welfare. For this, 18 beef calved cows were selected and divided into two groups: inseminated cows (AIG, n = 9), and not inseminated cows, the control group (CG, n = 9). Body condition score, uterus, and ovary evaluation were performed. Later, both groups were submitted into an estrus synchronization protocol and only the AIG group was inseminated. Blood components of urea, creatinine, AST, GGT, CK, glucose, triglycerides, cholesterol, HDL, LDL, VLDL, NEFA, BHB, cortisol, estradiol, progesterone, albumin, and total protein were measured 30 h before AI, and 4, 24, 48 and 168 h after AI. Statistical differences were considered when P <0.05. No differences between AIG and CG were observed. On the other hand, when the moment of insemination was evaluated, differences were observed for urea, creatinine, AST, GGT, CK, glucose, triglycerides, NEFA, BHB, albumin, and total protein. There was an oscillation of metabolic profiles depending on the time and procedures to which animals were exposed, even though it could be inferred that the AI process was incapable of altering those metabolic components on animals that were inseminated. Still, we can affirm that artificial insemination cannot be categorized as a negative reproduction tool on animal welfare. However, the containment and management procedures for AI may alter the metabolic profile of cows, especially the increase of CK.(AU)

O objetivo deste estudo foi verificar se o processo de inseminação artificial (IA) caracterizado como imobilização do animal, passagem do aplicador de sêmen pelo colo do útero e deposição do sêmen no útero, afetou o bem-estar de bovinos. Para tanto, foram selecionadas 18 vacas de corte paridas, divididas em dois grupos: grupo de animais inseminados (AIG, n = 9) e grupo de animais não inseminados, grupo controle (GC, n = 9). Foram avaliados o escore de condição corporal, útero e ovário. Posteriormente, ambos os grupos foram submetidos a um protocolo de sincronização de cio e apenas o grupo AIG foi inseminado. Componentes metabólicos como ureia, creatinina, AST, GGT, CK, glicose, triglicerídeos, colesterol, HDL, LDL, VLDL, NEFA, BHB, cortisol, estradiol, progesterona, albumina e proteína total foram mensurados 30 horas antes da IA e 4, 24, 48 e 168 horas após a IA. Diferenças estatísticas foram consideradas quando P <0,05. Não foram observadas diferenças entre os dois grupos, por outro lado, quando o momento da inseminação foi avaliado, diferenças foram observadas para ureia, creatinina, AST, GGT, CK, glicose, triglicerídeos, NEFA, BHB, albumina e proteína total. Houve uma variação dos perfis metabólicos em função do tempo e dos procedimentos que os animais foram submetidos, embora pode-se inferir que o processo de IA não foi capaz de alterar esses componentes metabólicos os animais inseminados. Ainda assim, observou-se que o processo de IA não foi categorizado como uma ferramenta negativa de reprodução com relação ao bem-estar animal. Porém, ainda assim, os procedimentos de contenção e manejo da IA podem alterar o perfil metabólico das vacas, principalmente o aumento da CK.(AU)

Effects of clinical mastitis and puerperal diseases on reproductive efficiency of dairy cows.

The objective of this study was to evaluate the effects of clinical mastitis (CM) occurring before or after the first AI postpartum, and puerperal diseases (PD) on the pregnancy per artificial insemination (P/AI), number of AI/conception, and days open (DO) of two different dairy herds (Girolando and Holstein). The CM, PD (retained placenta and metritis), and reproductive data were collected from two dairy farms throughout 1 year. Both farms were located in the southern region of Minas Gerais State, Brazil. One herd was composed of Girolando cows and the other of Holstein cows. Cows were inseminated after estrus detection or submitted to timed AI. Only CM cases (clots in milk accompanied or not by udder inflammation) that occurred before or after first AI postpartum (from calving until 35 days after the first AI) were considered. There were no effects of CM, PD, or both diseases on the reproductive efficiency of the Girolando herd. In the Holstein herd, a reduce P/AI and prolonged DO were verified for those affected by ≥ 2 CM cases. Holstein cows with CM also required more inseminations to become pregnant. A decrease in the P/AI and an increase in the number of AI/conception and DO were observed in cows of the Holstein herd that developed only CM, only PD, and for those diagnosed with both diseases. In summary, considering that some management differences exist between the two dairy farms, CM occurrence (before or after the first AI postpartum) and puerperal diseases negatively affected the reproductive efficiency of the Holstein herd. However, these diseases did not compromise the reproductive efficiency of the Girolando herd.

Non-infectious causes that increase early and mid-to-late pregnancy loss rates in a crossbreed dairy herd.

High reproductive performance is an element factor for profitability in dairy herds; although, pregnancy loss of non-infectious nature during early and mid-to-late embryonic period is increasing in dairy cattle. Based on that, the present study aimed to determine the incidence of pregnancy loss before and after 60 days of insemination, and to identify some non-infectious causes that could enhance it. The herd was composed by 600 crossbred dairy cows and those with a corpus luteum (CL) were treated with prostaglandinF2α, then inseminated, on the other hand, those without a CL were submitted to a timed artificial insemination protocol (TAI). Pregnancy losses rates were analyzed by logistic regression by SAS, and differences were considered significant when P < 0.05. The overall pregnancy loss and mid-to-late pregnancy loss were not affected by animal category (P > 0.05); although, early pregnancy loss was higher in cows than in heifers (11.90 vs. 3.39%). The early pregnancy loss was higher in those cows that calved on spring/summer when compared to those calved on fall/winter (9.22% vs. 16.11%), moreover, those inseminated during spring/summer tended to have higher early pregnancy loss when compared to those inseminated on fall/winter (13.35% vs. 8.57%). In conclusion, when some of non-infectious causes were evaluated, it was observed that cows that calved on spring/summer had higher pregnancy loss. At this point, strategies should be developing to minimize pregnancy loss in dairy herds, as it could be considered an important reproductive problem.

Mechanisms linking bacterial infections of the bovine endometrium to disease and infertility.

Bacterial infections of the endometrium after parturition commonly cause metritis and endometritis in dairy cattle, and these diseases are important because they compromise animal welfare and incur economic costs, as well as delaying or preventing conception. Here we highlight that uterine infections cause infertility, discuss which bacteria cause uterine disease, and review the evidence for mechanisms of inflammation and tissue damage in the endometrium. Bacteria cultured from the uterus of diseased animals include Escherichia coli, Trueperella pyogenes, and several anaerobic species, but their causative role in disease is challenged by the discovery of many other bacteria in the uterine disease microbiome. Irrespective of the species of bacteria, endometrial cell inflammatory responses to infection initially depend on innate immunity, with Toll-like receptors binding pathogen-associated molecular patterns, such as lipopolysaccharide and bacterial lipopeptides. In addition to tissue damage associated with parturition and inflammation, endometrial cell death is caused by a cholesterol-dependent cytolysin secreted by T. pyogenes, called pyolysin, which forms pores in plasma membranes of endometrial cells. However, endometrial cells surprisingly do not sense damage-associated molecular patterns, but a combination of infections followed by cell damage leads to release of the intracellular cytokine interleukin (IL)-1 alpha from endometrial cells, which then acts to scale inflammatory responses. To develop strategies to limit the impact of uterine disease on fertility, future work should focus on determining which bacteria and virulence factors cause endometritis, and understanding how the host response to infection is regulated in the endometrium.

Factors that enhance repeat breeder condition in a crossbred dairy herd submitted to reproductive synchronization protocols / Fatores que aumentam a condição de vacas repetidoras em um rebanho leiteiro mestiço submetido à protocolos de sincronização reprodutivos

The objective of this study was to evaluate the incidence of repeat breeder (RB) in crossbred dairy cows submitted to a timed artificial insemination (TAI) and estrous synchronization protocols. Data from 371 heifers and 997 lactating crossbred dairy cows were analyzed. Cows with more than 30 DPP with good uterine condition, no lameness or clinical mastitis, and body condition score ≥ 2.5 were used. The cows were divided between two groups, as: group 1: TAI ­ those cows with no CL at ultrasound exam that were enrolled in a TAI protocol and group 2: estrous synchronization ­ all those cows that had CL. The incidences of RB and factors relative to it were analyzed by logistic regression in SAS. The incidence of RB condition was higher in lactating cows than nonlactating heifers (P < 0.001). There was a tendency of RB condition to higher in cows that calved during spring/summer than those calved at autumn/winter season. The parturition condition had no effect on RB incidence (P > 0.001); however, lactation degree influenced the incidence of RB (P < 0.001). Repeat breeder condition was greater in lactating crossbred dairy cows than in heifers. Moreover, the presence of RB cows was higher in those from second parity group when compared to those from first, third and four parity groups; on the other hand, parturition condition did not influence the appearance of RB cows in the herd.(AU)

O objetivo deste estudo foi avaliar a incidência vacas repetidoras (VR) leiteiras mestiças submetidas à inseminação artificial em tempo fixo (IATF) e protocolos de sincronização de estro. Dados de 371 novilhas e 997 vacas leiteiras mestiças foram analisados. Vacas com mais de 30 dias pós-parto, boa condição uterina, sem mastite clínica e com escore corporal ≥ 2,5 foram utilizadas. As vacas foram divididas em dois grupos, sendo: grupo 1: IATF ­ vacas sem corpo lúteo no exame ultrassonográfico e que foram submetidas ao protocolo de IATF, e grupo 2: sincronização de cio ­ aquelas vacas com corpo lúteo presente. A incidência de VR e seus fatores foram analisados por regressão logística pelo SAS. A incidência de VR foi maior em vacas em lactação em relação às fêmeas não-lactantes (P < 0,001). Ainda, uma tendência de um maior número de VR foi observada naquelas que pariram na primavera/verão quando comparadas às paridas no outono/inverno. O tipo de parto (normal vs. não normal) não influenciou na incidência de VR (P > 0,001), no entanto, foi detectado efeito do número de partos (P > 0.01). Dessa forma, nas categorias animais avaliadas, a incidência de VR foi maior em vacas leiteiras mestiças do que em novilhas. Uma tendência de VR foi observada naquelas que pariram na primavera/verão quando comparadas aquelas que pariram no outono/inverno, ainda a incidência de VR foi maior naquelas de segunda parição quando comparadas as de primeira, terceira ou quarta, ao passo que o tipo de parto (normal vs. não normal) não influenciou sobre a incidência de VR.(AU)

Incidence of subclinical endometritis and its effects on reproductive performance of crossbred dairy cows.

In dairy cattle, uterine infections are not life threatening and often unavoidable; however, they reduce fertility and increase the production costs of properties. The aim of this study was to evaluate the incidence of subclinical endometritis from 32 to 70 days in milk (DIM) and its effects on the reproductive performance of crossbred dairy cows. Lactating cows (Holstein/Gir; n = 172), with no history of retained placenta, without clinical signs of uterine infection were used. The body condition score (BCS) was evaluated on a scale from 1 to 5. Ultrasound examination was performed to evaluate uterine lining and ovarian activity, while vaginal mucus was analyzed by gloved hand. The diagnosis of subclinical endometritis was performed by endometrial cytobrush technique. The samples were collected, stained, and examined microscopically; positive cases for subclinical endometritis were considered with the presence of ≥5 % of neutrophils. Later, the cows were submitted to conventional artificial insemination or timed artificial insemination. The incidence of subclinical endometritis in the herd was 26 %, and this was not affected by the season of calving, presence of corpus luteum, DIM, and parity. Cows with a BCS ≤2.50 had a higher incidence of subclinical endometritis. The conception rate to first insemination and pregnancy rate at 150 days postpartum were not influenced by the presence of subclinical endometritis in crossbred dairy cows.

Exploratory analysis of differences in sperm morphology in Nelore and Gir (Bos indicus) bulls.

The aim of this study was to evaluate the effects of breed and season on semen quality parameters of zebu bulls. Data (1,632 registers) of semen production from Gir (n = 4) and Nelore (n = 15) bulls were collected between October 2005 and November 2009. The ejaculates were collected twice a week during various seasons (summer, fall, winter, and spring) and evaluated for the following semen parameters: ejaculate volume, sperm concentration, sperm motility, forward progressive motility (FPM), and sperm morphology. Factor analysis was used to determine the relationship among variables. The effect of breed (Gir and Nelore) and season and their cross effect on each parameter and extracted factor were tested using ANOVA. A negative correlation (P < 0.05) was observed between FPM and proximal droplet, as well as with abnormal loose head, abnormal small head, pouch formation, abnormal mid-piece, and strongly folded tail. Gir bull sperm showed more major defects, detached acrosome, and minor FPM (P < 0.01), whereas Nelore bulls showed a higher number of sperm with normally loose head.

Timed artificial insemination and early diagnosis of pregnancy to reduce breeding season in Nelore beef cows.

The purpose of this study was conducted to evaluate the use of a system of timed artificial insemination (TAI) and early diagnosis of pregnancy in Nelore beef cows in order to reduce the breeding season (BS) from 150 to 90 days and analyze the factors affecting the conception rate of cows selected for TAI. The study included 110 Nelore cows belonging to the Experimental Farm Capim Branco, Brazil. The body condition scores (BCS) were evaluated, and those above 2.50 were included in the TAI protocol. Five groups of cows were formed at intervals of 21 days postpartum and then commenced a 9-day progesterone protocol. Progesterone devices were used three times. Statistical analysis of binary variables was performed by logistic regression in the program Proc Logistic of Sas. A 5% of significance (P < 0.05) was used to determine the presence of statistical differences and the effect of independent variables on conception rate. The TAI and early diagnosis of pregnancy reduces the BS from 150 to 90 days without changing the pregnancy rate. The conception rate was not affected by the presence of the calf (P > 0.05), the corpus luteum (P > 0.05), the BCS (P > 0.05), or the number of reuses of the intravaginal progesterone device (P > 0.05). Therefore, this technique is beneficial for breeding Nelore beef cattle and can be used in anestrous cows with a BCS above 2.50. Additionally, the progesterone devices can be reused to reduce the cost of the protocol without compromising the results.

Eficiência do Estrotect® na identificação de estros em vacas leiteiras mestiças / Estrotect® efficiency in identifying estrus in crossbred dairy cows

Dentre os problemas que diminuem a eficiência reprodutiva em rebanhos leiteiros a falha de detecção de estro é um dos fatores mais importantes. Objetivou-se avaliar a eficiência de um dispositivo que auxilia a detecção de estros – Estrotect® e compará-lo com a detecção visual em 58 vacas mestiças leiteiras Holandês/Gir com produção média de 18,5 kg de leite/dia/vaca. Os animais foram distribuídos em dois grupos: o grupo IATF (n= 21) foi submetido a um protocolo de inseminação artificial em tempo fixo (IATF) e 10 dias após a inseminação foi fixado o dispositivo nos animais. No grupo PG (n=37), foi administrado prostaglandina 25mg/animal/IM e imediatamente colado o adesivo. Essas vacas foram monitoradas por meio de exame ultrassonográfico num período de 28 dias para verificação do desenvolvimento do corpo lúteo. Para verificar se ocorreram alterações na eficiência do dispositivo quanto aos grupos utilizou-se a regressão logística e para comparar a eficiência do mesmo em relação à observação visual utilizou-se o teste de MCNemar. Não foi detectado efeito de grupo (P>0,05) na eficiência da detecção visual de estro ou da ferramenta auxiliar. Também não foi detectada diferença entre a eficiência da detecção visual de estro e o dispositivo (P>0,05). A ferramenta auxiliar de detecção de estro (Estrotect®) se mostrou tão eficiente quanto a detecção visual e pode ser utilizada em programas de inseminação artificial de vacas leiteiras.

Among the problems that reduce the reproductive efficiency in dairy cows the failure in estrus detection is a major factor. The objective of this study was to evaluate the efficiency of Estrotect®, a device that aids heat detection in cattle, and to compare it with visual detection in 58 crossbred Holstein/Gyr dairy cows producing 18.5Kg/milk/cow. The animals were divided into two groups: the TAI (n = 21) underwent a fixed-time artificial insemination protocol (TAI), and 10 days after insemination, the device was fixed to the cows. In the PG group (n = 37) prostaglandin (dinoprost tromethamine) 25mg/animal/IM was administered, and then the device was put in place. These cows were then monitored by ultrasound over a period of 28 days to verify the development of the corpus luteum. Logistic regression was used to find out if there were alterations in the efficiency of Estrotect® within the groups, and the McNemar test was used to compare its efficiency to that of visual observation. No group effect was detected (P> 0.05) on the efficiency of visual detection nor on that of the auxiliary device. Neither was there observed any difference between the efficiency of visual detection of estrus and that of the device (P> 0.05). The auxiliary tool for the detection of estrus, (Estrotect ®), proved to be just as effective as visual detection. Thus, it can be used as an auxiliary device in conjunction with visual observation, and also as a unique tool for the detection of estrus in order to simplify the management of large herds.