Effect of recombinant bovine somatotropin (rbST) treatment on follicular population and development in non-lactating dairy cows

The aim of this study was to evaluate the long term effects of recombinant bovine somatotropin (rbST) on follicle population and ovulatory follicle development in non-lactating dairy cows. Twenty-one Jersey cows were allocated in rbST (n=11) or control (n=10) groups. On day -60, cows in rbST group received 500 mg of somatotropin (s.c. Lactotropin, Elanco). On day 0, control and rbST cows received an intravaginal progesterone-releasing device (1.9 g, CIDR, Zoetis) and GnRH (100 mg, IM, Factrel, Zoetis). On day 8, cows received PGF2α (25 mg, IM, Lutalyse, Zoetis) and the CIDR was removed. Twelve hours after device removal (D8), serum, follicular fluid and granulosa cells samples were collected. Serum and follicular concentration of estradiol (E2) and progesterone (P4) were analyzed. Total RNA was extracted from granulosa cells to measure gene expression of LHCGR, STAR, HSD-3B1, CYP11A1, CYP19A1, CYP17A1, IGFR and PAPPA by real time PCR. Ultrasonography was performed on days -60, -53, -46, -14, -7, 0 and 8 for antral follicle count. Results were analyzed by repeated measures ANOVA and t-test. There was no effect of rbST treatment on the number of follicles during the 60 days period, as well as no effect on serum and follicular fluid E2 and follicular fluid P4 at the moment of follicle aspiration. There was a reduction in PAPPA (P = 0.006), CYP11A1 (P = 0.04) and CYP19A1 (P = 0.002) mRNA levels in granulosa cells of the pre ovulatory follicle of rbST treated cows. In conclusion, a single dose of rbST did not have long term effects on antral follicle population, serum and follicular E2/P4 concentrations in non-lactating dairy cows. Despite that, rbST injection decreased granulosa cell expression of genes related to steroidogenesis in the pre-ovulatory follicle

Salmonella spp. Isolated by Miniaturized Most Probable Number and Conventional Microbiology in Poultry Slaughterhouses

Background: Salmonella spp. are frequently isolated from fowls, and their detection in poultry products varies according to the breeding system and the slaughtering process, bringing risks to the consumer and compromising the marketability. The control of Salmonella in poultry slaughterhouses is based on the detection of bacteria, but the quantification of the agent would be important in assessing risk, as well as in obtaining data to determine the capacity of each step of the process to decrease or increase bacterial contamination. The aims of this study were to propose a method for the quantification of Salmonella in poultry slaughterhouses, frequency of isolation and serovars identified.Materials, Methods & Results: Twenty-one broiler flocks from seven federally inspected slaughterhouses in southern Brazil, totaling 1,071 samples, were assessed by miniaturized most probable number (mMPN) and conventional microbiology. The samples were collected in triplicate at 17 points, which included cloacae, transportation cages before and after sanitization, water (scald tank, supply, pre-chiller and chiller), and carcasses (before and after scalding, defeathering, rinsing, evisceration, final rinsing, chilling at 4ºC, and freezing at -12°C for 24 h, 30 and 60 days). Typical Salmonella colonies were submitted to TSI, LIA, SIM, urea, and polyvalent anti-O antiserum tests, and to final identification by Microarray by Check&Trace. Nine of the 1,071 (0.83%) samples analyzed by mMPN and by conventional microbiology were positive for Salmonella and the following serovars were identified: Anatum, Brandenburg, Agona, Tennessee, Bredeney, Schwarzengrund and Infantis.Discussion: This positive rate was lower than that described by other authors, whose rates ranged from 3% and 39% for the isolation of Salmonella spp. from different sources, such as slaughterhouses and retail sales in samples collected in Brazil. The low frequency of isolation of Salmonella in this study can be attributed to the efficiency of control systems used from the field to the slaughterhouse, such as Good Manufacturing Practices (GMP) and Sanitation Standard Operating Procedures (SSOP), which are HACCP requirements. Also, when slaughtering technology actions are properly managed, such as water replacement and temperatures lower than 4ºC in the chiller, the initial contamination by Salmonella spp. can be reduced, with a decline in contamination from 70% to 20%, and with a reduction in the contamination of broiler carcasses after chilling from 15.8% to 3.3%. On the other hand the contamination of carcasses by Salmonella before pre-chilling and in post-chilling might be due to the automated system, inadequate temperatures during chilling, and inappropriate water chlorination in the assessed meat-packing plant. Of the 17 points evaluated, seven were positive for Salmonella, especially the cages after sanitization and frozen carcasses. The contamination by Salmonella spp. in transportation cages after sanitization indicates inefficiency of the automated system as well as possible bacterial resistance to the sanitizers used in SSOP while the isolation in carcasses frozen for 24 h and 60 days demonstrates the thermal resistance of the bacterium to a conservation method widely used in the food industry. In this work, just one of the nine positive samples for Salmonella was identified by conventional methods (CM) and mMPN. The discrepancy between methods can be explained by the heterogeneous distribution of Salmonella and other bacteria in naturally contaminated samples. Samples that were positive in the qualitative test but negative in the mMPN protocol could have had a number of Salmonella below the detection amount

J-Synch protocol associated with estrus detection in beef heifers and non-lactating cows

The aim of this study was to compare the J-Synch and conventional protocols associated with estrus detection in beef heifers and to compare pregnancy rate between non-lactating cows displaying estrus or not during the J-Synch protocol. In Experiment 1, heifers were subjected to timed artificial insemination (AI) in a conventional protocol with ECP (n=147) or J-Synch protocol plus eCG (n=149). The AI occurred 12 hours after estrus expression; or 48 (Conventional protocol) and 72 hours (J-Synch protocol) after device removal for animals not displaying estrus. The J-Synch group received 10 µg of GnRH at AI. In Experiment 2, the JSynch was performed (n=116 cows), but without eCG injection, and estrus was monitored. Pregnancy rate was not different between protocols in Experiment 1 (Conventional: 50.68%; J-Synch: 60.4%). Heifers that displayed estrus had higher pregnancy rate only in the conventional protocol. In Experiment 2, pregnancy rate was not different between cows that displayed estrus or not. Therefore, performing AI earlier according to estrus expression increases pregnancy rate in conventional protocol, however it does not increase pregnancy rate in the J-Synch protocol

Supplementation of in vitro maturation medium with serum from dairy cows at early and late lactation in the in vitro production of bovine embryos

The transition period of dairy cows is characterized by several metabolic alterations, due to the negative energy balance, which negatively affects fertility. The aim of this study was to evaluate the effect of supplementing the in vitro maturation medium with serum from dairy cows at early and late lactation in the oocyte maturation on the rates of nuclear maturation, cleavage and initial embryo development. The cumulus-oocyte complexes (COCs) were randomly divided into treatments: LL: serum from four cows at 354 ± 3.8 days in milk (DIM); and EL: serum from four cows at 10.8 ± 4.8 DIM. Subsequently, the COCs were submitted to maturation, fertilization; and in vitro culture in an incubator at 39°C with 5% CO2. For analysis of nuclear maturation; oocytes were collected after IVM, denuded and stained with Hoechst®. No difference was observed in the number of oocytes that reached metaphase II among treatments. The cleavage rate was not different between LL (59.3%; 320/540) and EL (47.0%; 243/517) treatments (P>0.05), as embryonic development between LL (15.0%; 81/540) and EL (16.6%; 86/517) (P>0.05). It was concluded that supplementation with serum from cows at different lactation periods during the bovine oocyte IVM did not influence the results of in vitro embryo production

Relationship between follicular size, ovulation time and pregnancy rate of cattle submitted to conventional or in block FTAI

The aim of study 1 was to evaluate the relationship between follicular size (FS) and moment of ovulation in Bos taurus cows. Data of 138 cows from 4 protocol of estrus synchronization were used. Cows received an intravaginal progesterone-releasing device plus 2 mg of Estradiol Benzoate (EB) on D0. On D8, device was removed and cows were randomized in four groups: Estradiol cipionate (ECP, n=76), administrated 48 hours after device removal; EB (n=19) administrated after 24 hours; GnRH (n=28) administrated after 60 hours; or Control (no inductor; n=15). Ultrasonography evaluations were performed each 12 hours after device removal to measure FS and to detect ovulation. In ECP group, cows with larger FS had earlier ovulation (R²=0.96, P 14 mm demonstrated higher pregnancy per AI (P/IA) than cows with FS <10 mm or < 11 mm (dairy and beef cows, respectively; P<0.05). The FS was correlated to luteal size (R²=0.71) and cows with larger corpus luteum had higher P/AI (P<0.05). The aim of study 3 was to evaluate serum estradiol and progesterone concentrations. Serum samples of 14 cows from Study 1 were used. Cows with larger LS had lower serum progesterone at ovulation. The aim of Experiment 2 was to evaluate whether chancing AI time according FS would improve P/AI. Dairy (n=65) and beef cows (n=90) were synchronized as described for ECP group. Beef cows received 350 IU of eCG on D8. Ultrasonography evaluation was performed at AI and cows were divided according FS in two groups (≤ 10.4 mm or ≥ 10.5 mm) and after randomized in control group (Conventional FTAI), Block 1 (≥ 10.5 mm): AI 48 hours after device removal; or Block 2 (≤ 10.4 mm): AI 65 hours after device removal. No differences were observed between Conventional and blocks FTAI. Lower pregnancy rate observed in cows with smaller follicles possibly was not attributed with asynchrony between AI and moment of ovulation.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESO objetivo do estudo 1 foi avaliar a relação do diâmetro folicular (DF) e momento da ovulação em fêmeas Bos taurus. Dados de 138 fêmeas bovinas de 4 protocolos de sincronização foram utilizados. Os animais receberam dispositivo intravaginal de progesterona (P4) e 2 mg de Benzoato de Estradiol (BE) no D0. No D8, a P4 foi retirada e as fêmeas foram divididas aleatoriamente em 4 grupos: Cipionato de Estradiol (ECP; n=76), administrado na retirada da P4; BE (n=19), administrado 24 horas após; GnRH (n=28), administrado 60 horas após; ou Controle (n=15). Foram realizadas avaliações ultrassonográficas de 12 em 12 horas a partir do D8 para a mensuração do DF e detecção da ovulação. No grupo ECP, vacas com maior DF, apresentam ovulação mais precoce (R²=0,96, P<0,0001). O estudo 2 foi realizado para avaliar relação do DF, volume do corpo lúteo (CL) e prenhez. Foi realizada inseminação artificial (IA) em 30 fêmeas leiteiras do grupo ECP do estudo 1 e em 83 fêmeas de corte sincronizadas conforme descrito no grupo BE. Dados de avaliações ultrassonográficas no momento da IA, 7 e 30 dias após a IA foram utilizados. Vacas com DF >14 mm apresentaram maior prenhez/IA (P/IA) do que vacas com DF <10mm ou < 11mm (leite e corte, respectivamente; P<0,05). O DF foi correlacionado com o volume do CL (R²=0,71) e maiores CLs apresentaram maior P/IA (P<0,05). O objetivo do estudo 3 foi avaliar as concentrações séricas de E2 e P4 nos animais do grupo ECP. Avaliações séricas de 14 animais do Estudo 1 foram utilizadas nessa avaliação. Vacas com maiores DF apresentaram maior concentração de P4 no momento da ovulação. O objetivo do Experimento 2 foi avaliar se a alteração do momento da IA de acordo com o DF aumenta a P/IA de bovinos. Foram utilizadas 65 fêmeas leiteiras e 90 de corte sincronizadas conforme descrito para o grupo ECP. Fêmeas de corte receberam 350 UI de eCG no D8. Avaliação ultrassonográfica foi realizada no momento da IA e as fêmeas foram divididas de acordo com o DF em dois grupos (≤ 10,4 mm ou ≥ 10,5 mm) e após divididas aleatoriamente em grupo controle (IATF convencional) e bloco 1 (≥ 10,5 mm): IA 48 horas após remoção da P4); ou bloco 2 (≤ 10,4 mm): IA 65 horas após remoção da P4). Não houve diferença na P/IA das fêmeas submetidas a IATF convencional ou em blocos. A menor taxa de prenhez encontrada nas fêmeas com menores folículos possivelmente não foi atribuída a assincronia entre momento da IA e ovulação

Sympathetic nerve activity is decreased during device-guided slow breathing

It is known that slow breathing (<10 breaths min(-1)) reduces blood pressure ( BP), but the mechanisms involved in this phenomenon are not completely clear. The aim of this study was to evaluate the acute responses of the muscle sympathetic nerve activity, BP and heart rate (HR), using device-guided slow breathing ( breathe with interactive music (BIM)) or calm music. In all, 27 treated mild hypertensives were enrolled. Muscle sympathetic nerve activity, BP and HR were measured for 5min before the use of the device (n=14) or while subjects listened to calm music (n=13), it was measured again for 15 min while in use and finally, 5min after the interventions. BIM device reduced respiratory rate from 16 +/- 3 beats per minute (b.p.m) to 5.5 +/- 1.8 b.p.m (P<0.05), calm music did not affect this variable. Both interventions reduced systolic (-6 and -4mmHg for both) and diastolic BPs (-4mmHg and -3mmHg, respectively) and did not affect the HR (-1 and -2 b.p.m respectively). Only the BIM device reduced the sympathetic nerve activity of the sample (-8bursts min(-1)). In conclusion, both device-guided slow breathing and listening to calm music have decreased BP but only the device-guided slow breathing was able to reduce the peripheral sympathetic nerve activity. Hypertension Research ( 2010) 33, 708-712; doi: 10.1038/hr.2010.74; published online 3 June 201

Effects of long term device-guided slow breathing on sympathetic nervous activity in hypertensive patients: a randomized open-label clinical trial

Purpose: Device-guided slow breathing (DGB) is indicated as nonpharmacological treatment for hypertension. The sympathetic nerve activity (SNA) reduction may be one of the mechanisms involved in blood pressure (BP) decrease. The aim of this study is to evaluate the long-term use of DGB in BP and SNA. Subjects and methods: Hypertensive patients were randomized to listen music (Control Group–CG) or DGB (aim to reduce respiratory rate to less than 10 breaths/minute during 15 minutes/day for 8 weeks). Before and after intervention ambulatory blood pressure monitoring (ABPM), catecholamines and muscle sympathetic nerve activity (MSNA) by microneurography were performed. Results: 17 volunteers in the DGB and 15 in the CG completed the study. There was no change in office BP before and after intervention in both groups. There was a reduction in systolic and diastolic BP in the awake period by ABPM only in the CG (131 ± 10/92 ± 9 vs 128 ± 10/88 ± 8mmHg, p < 0.05). In relation to SNA, no difference in catecholamines was observed. In the volunteers who had a microneurography record, there was no change the MSNA (bursts/minute): DGB (17(15–28) vs 19(13–22), p = 0.08) and CG (22(17–23) vs 22(18–24), p = 0.52). Conclusion: Long-term DGB did not reduce BP, catecholamines levels or MSNA in hypertensive patients. ClinicalTrials.gov identifier: NCT0139072

Salmonella spp. Isolated by Miniaturized Most Probable Number and Conventional Microbiology in Poultry Slaughterhouses

Background: Salmonella spp. are frequently isolated from fowls, and their detection in poultry products varies according to the breeding system and the slaughtering process, bringing risks to the consumer and compromising the marketability. The control of Salmonella in poultry slaughterhouses is based on the detection of bacteria, but the quantification of the agent would be important in assessing risk, as well as in obtaining data to determine the capacity of each step of the process to decrease or increase bacterial contamination. The aims of this study were to propose a method for the quantification of Salmonella in poultry slaughterhouses, frequency of isolation and serovars identified.Materials, Methods & Results: Twenty-one broiler flocks from seven federally inspected slaughterhouses in southern Brazil, totaling 1,071 samples, were assessed by miniaturized most probable number (mMPN) and conventional microbiology. The samples were collected in triplicate at 17 points, which included cloacae, transportation cages before and after sanitization, water (scald tank, supply, pre-chiller and chiller), and carcasses (before and after scalding, defeathering, rinsing, evisceration, final rinsing, chilling at 4ºC, and freezing at -12°C for 24 h, 30 and 60 days). Typical Salmonella colonies were submitted to TSI, LIA, SIM, urea, and polyvalent anti-O antiserum tests, and to final identification by Microarray by Check&Trace. Nine of the 1,071 (0.83%) samples analyzed by mMPN and by conventional microbiology were positive for Salmonella and the following serovars were identified: Anatum, Brandenburg, Agona, Tennessee, Bredeney, Schwarzengrund and Infantis.Discussion: This positive rate was lower than that described by other authors, whose rates ranged from 3% and 39% for the isolation of Salmonella spp. from different sources, such as slaughterhouses and retail sales in samples collected in Brazil. The low frequency of isolation of Salmonella in this study can be attributed to the efficiency of control systems used from the field to the slaughterhouse, such as Good Manufacturing Practices (GMP) and Sanitation Standard Operating Procedures (SSOP), which are HACCP requirements. Also, when slaughtering technology actions are properly managed, such as water replacement and temperatures lower than 4ºC in the chiller, the initial contamination by Salmonella spp. can be reduced, with a decline in contamination from 70% to 20%, and with a reduction in the contamination of broiler carcasses after chilling from 15.8% to 3.3%. On the other hand the contamination of carcasses by Salmonella before pre-chilling and in post-chilling might be due to the automated system, inadequate temperatures during chilling, and inappropriate water chlorination in the assessed meat-packing plant. Of the 17 points evaluated, seven were positive for Salmonella, especially the cages after sanitization and frozen carcasses. The contamination by Salmonella spp. in transportation cages after sanitization indicates inefficiency of the automated system as well as possible bacterial resistance to the sanitizers used in SSOP while the isolation in carcasses frozen for 24 h and 60 days demonstrates the thermal resistance of the bacterium to a conservation method widely used in the food industry. In this work, just one of the nine positive samples for Salmonella was identified by conventional methods (CM) and mMPN. The discrepancy between methods can be explained by the heterogeneous distribution of Salmonella and other bacteria in naturally contaminated samples. Samples that were positive in the qualitative test but negative in the mMPN protocol could have had a number of Salmonella below the detection amount