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 pentoxifylline supplementation to semen extender on post-breeding inflammation response assessed by endometrial cytology and vascular perfusion in mares.

The aim was to evaluate effects of addition of pentoxifylline to skimmed milk semen extender on uterine inflammatory response. Thirty-six estrous cycles of 15 mares were randomly divided into five groups for artificial insemination (AI): Control: mimicking the AI procedure (n = 7); Extender: deposition of skimmed milk based extender (n = 7); Extender + PTX: skimmed milk based extender plus pentoxifylline (7.18 mM; n = 8); Semen: semen diluted with extender without pentoxifylline (n = 7), and Semen + PTX: semen diluted with extender containing pentoxifylline (n = 7). Mares in estrus were examined by trans-rectal palpation and using ultrasonography, and ovulation was induced. Uterine hemodynamics were assessed immediately before ovulation induction (T-30), immediately before AI (T0), 2 (T2), 6 (T6), 12 (T12), 24 (T24) and 48 (T48) h after AI. Endometrial samples were collected 6 h after AI, and slides were stained and examined to determine percentage of PMN. Pentoxifylline had no additional effect on vascular perfusion. There was a major inflammatory response with pentoxifylline treatment that was greater than that of the control group. In the group treated with Extender + PTX, there were more PMN (57.98 ±â€¯9.42%) than in the group treated with Extender (20.20 ±â€¯6.63%) and in the Semen + PTX group more PMN (82.84 ±â€¯5.71%) than in the Semen-treated group (47.83 ±â€¯10.61%). These findings indicate the addition of pentoxifylline does not stimulate blood flow; however, it induces a greater immune defense response because more neutrophils migrate to the uterine lumen.

Recovery of normal testicular temperature after scrotal heat stress in rams assessed by infrared thermography and its effects on seminal characteristics and testosterone blood serum concentration.

Reestablishment of testicular normal temperature after testicular heat stress is unknown and its effect varies widely. The aim of this study was to investigate the impact of scrotal insulation (IN) on testicular temperature and its relation to semen quality and testosterone blood serum concentration. For this, 33 rams were used; 17 submitted to IN for 72 hours (using bags involving the testes) and 16 not submitted to IN (control group). The experiment was performed between August and December 2013 in Pirassununga, Brazil (21°56″13″ South/47°28'24″ West). Seminal characteristics, testosterone blood serum concentration, rectal temperature (RT), respiratory frequency, scrotal superficies mean temperature (SSMT), and eye area mean temperature (EAMT) were analyzed 7 days before IN and 21, 35, 49, 63, and 90 days afterward. Scrotal superficies mean temperature and EAMT were measured by thermography camera FLIR T620. Testosterone was evaluated by radioimmunoassay. Analysis of variance was used to determine the main effects of treatment, time, and treatment-by-time interaction using PROC MIXED of SAS software adding command REPEAT. Pearson correlation test was used to verify correlation between SSMT, EAMT, RT, and respiratory frequency. Significant difference was considered when P ≤ 0.05. At the end of IN, SSMT was higher (P < 0.05) in insulated group (32.26 ± 0.19(o)C) than in control group (30.58 ± 0.18(o)C), and the difference between rectal and testicular (deduced from SSMT) temperatures was 1.12 °C; in the other times of the evaluation this difference was between 2.91 and 4.25 °C in IN group. Scrotal superficies mean temperature was reestablished 24 hours after IN. Rectal temperature and EAMT presented correlation (r = 0.59; P < 0.0001). There was time-by-treatment interaction for total sperm (P = 0.0038) and progressive motility (P = 0.01), abnormal spermatozoa (P < 0.0001), membranes integrity (P < 0.0001), induced thiobarbituric acid reactive substances (TBARSs; P = 0.05), and DNA integrity (P = 0.0004). These semen characteristics were negatively affected 21 days after IN, and excluding induced TBARSs and abnormalities, recovered 35 days afterward; induced TBARSs just were affected after 49 days of IN; sperm abnormalities just recovered after 63 days. Testosterone blood serum concentration was lesser in insulated rams (P = 0.03). Thus, the difference of 1.12 °C between RT and testicular temperature impacts semen quality and testosterone blood serum concentration. Moreover, this study shows that rams can recover testes temperature efficiently toward IN and that infrared thermography is an efficient tool to identify differences on SSMT.

Low-level laser therapy to recovery testicular degeneration in rams: effects on seminal characteristics, scrotal temperature, plasma testosterone concentration, and testes histopathology.

The aim of this study was to investigate the efficiency of low-level laser therapy (LLLT) to recovery testicular degeneration in rams. In the first study, rams were induced to testicular degeneration by scrotal insulation, and then, they were treated using LLLT at 28 J/cm(2) (INS28) or 56 J/cm(2) (INS56) energy densities. Sperm kinetics, morphology, and membranes integrity as well as proportion of lumen area in seminiferous tubule were assessed. In the second study, rams were submitted or not to scrotal insulation and treated or not by the best protocol of LLLT defined by experiment 1 (INS28). In this study were evaluated sperm kinetics, morphology, membranes integrity, ROS production, and DNA integrity. Testosterone serum concentration and proportion of lumen area in seminiferous tubule were also analyzed. Insulation was effective in promoting sperm injuries in both experiments. Biostimulatory effect was observed in experiment 1: INS28 presented smaller proportion of lumen area (P = 0.0001) and less degeneration degree (P = 0.0002). However, in experiment 2, there was no difference between the groups (P = 0.17). In addition, LLLT did not improve sperm quality, and there was a decreasing for total and progressive motility (P = 0.02) and integrity of sperm membranes (P = 0.01) in LLLT-treated groups. Moreover, testosterone concentration was not improved by LLLT (P = 0.37). Stimulation of aerobic phosphorylation by LLLT may have led to a deregulated increase in ROS leading to sperm damages. Thus, LLLT at energy of 28 J/cm(2) (808 nm of wavelength and 30 mW of power output) can induce sperm damages and increase the quantity of cells in seminiferous tubule in rams.

Uterine lavage is efficient to recover endometrial cytology sample and does not interfere with fertility rate after artificial insemination in cows.

Productivity rates directly depend on the fertility of a herd, which in turn can be influenced by many factors. Semen deposited in the female reproductive tract is foreign to the body and, in response to this invasion, produces an inflammatory reaction, which is characterized by rapid infusion of polymorphonuclear (PMN) cells. Techniques to obtain an endometrial sample are usually invasive and can mask the true inflammatory response. Ultrasound is a noninvasive technique and can contribute to the diagnosis of postartificial insemination (AI) inflammatory response in cattle. The present study was divided into two experiments. The aim of experiment 1 was to compare two methods of endometrial cytology collection, uterine cytobrush (UC) and uterine lavage (UL), and their effects on uterine hemodynamics that provide information about blood flow. The two methods were evaluated by Doppler ultrasound using the spectral and color modes. For that purpose, 19 Nellore cows were synchronized for timed AI and subjected to UC (n = 9) or UL (n = 10). The techniques were performed 4 hours after AI. The results showed that both techniques allow collection of a good quality sample and with enough PMN cells to perform counting. More PMN cells were obtained by UL than UC. There was no difference in uterine blood flow between the UC and UL groups in any of the periods evaluated (34 hours before and 4, 24, and 48 hours after collection of uterine sample). On the basis of results of experiment 1, the effect of UL on fertility was studied in experiment 2. A total of 128 Nellore cows were synchronized for TAI; 35 cows were subjected to endometrial cytology by UL 4 hours after AI, and 93 were not submitted to any procedure (control). Pregnancy diagnosis was performed by transrectal ultrasound 30 days after AI. Pregnancy rates did not differ between UL (54.29%) and control (56.99%) groups. The results of this study showed that UL allows the collection of more representative cells of the surface of the uterus than UC technique and causes no damage to the reproductive tract. Moreover, UL did not affect pregnancy rate when performed 4 hours after AI.

Correlations between testicular hemodynamic and sperm characteristics in rams / Correlações entre a hemodinâmica testicular e as características espermáticas em carneiros

The testicular artery is responsible for the blood supply that reaches the testis and has great importance in heat radiation. Vascular changes in the testis may lead to damage in sperm production, reflected in sperm motility and morphology. The aim of the present study was to evaluate correlations between testicular vascularity and sperm characteristic. Eight adult Santa Ines rams showing different reproductive status were used. The testicular vascularity and sperm characteristics were evaluated fortnightly during 90 days. Color Doppler ultrasonography was used to evaluate the testicular hemodynamic. Resistance index (RI) and pulsatility index (PI) of the testicular artery were evaluated by spectral-Doppler mode. The color-Doppler mode was used to evaluate the blood flow of the pampiniform plexus and testicular parenchyma. The semen analyses assessed were volume, concentration, motility, and morphology. The data were submitted to Pearson´s linear correlations test (p 0.05 was considered significant). No correlations were found between motility and testicular hemodynamic. The percentage of total sperm defects was positively correlated to left and right parenchymal score and to left RI and PI. On the other hand, the pampiniform plexus score was positively correlated to the number of colored pixels and negatively correlated to the RI and PI, for both sides. This study showed that the increase of sperm defect can be related to increase of testicular blood flow; however, more studies are need.

A artéria testicular é responsável pelo fluxo de sangue que chega aos testículos e tem grande importância na termorregulação. Mudanças vasculares nos testículos podem levar à queda da produção espermática, refletindo na motilidade e morfologia. O objetivo deste trabalho foi avaliar as correlações entre a vascularização testicular e as características espermáticas. Foram utilizados oito carneiros adultos Santa Inês com diferentes status reprodutivos. A vascularização testicular e as características seminais foram avaliadas por um período de 90 dias. A ultrassonografia Doppler colorida foi utilizada para avaliar a hemodinâmica testicular. Foram calculados os índices de resistência (RI) e os índices de pulsatilidade (PI) com o modo espectral do Doppler. O modo colorido do Doppler foi utilizado para analisar o fluxo sanguíneo do plexo pampiniforme e do parênquima testicular. As características seminais avaliadas foram o volume, concentração, motilidade e morfologia. Os dados foram submetidos ao teste de correlação linear de Person (P 0.05 foi considerado significativo). Não foram encontradas relações entre motilidade e a hemodinâmica testicular. A porcentagem de defeitos totais correlacionou-se positivamente com o escore de vascularização dos parênquimas direito e esquerdo, e com o RI e PI esquerdos. Também o escore de vascularização dos plexos se correlacionou positivamente com a média de pixels e negativamente com o RI e PI, de ambos os lados. Este trabalho mostrou que o aumento de defeitos espermáticos pode estar correlacionado com o aumento do fluxo sanguíneo nos testículos; contudo, mais estudos são necessários.