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Bacillus spp. supplementation promotes feed efficiency in mid- to late-lactation dairy cows and affects rumen fermentation traits of rumen-fistulated females offered a corn silage–based total mixed ration diet 在以玉米青贮为基础的全混合日粮中,添加芽孢杆菌可提高泌乳中后期奶牛的饲料效率,并影响瘤胃瘘母奶牛的瘤胃发酵特性
Pub Date : 2025-07-01 DOI: 10.3168/jdsc.2024-0686
Bruno A.V. Arthur , Luiz Gustavo Nussio , Oscar C.M. Queiroz , Greicieli de Morais , Rafael H.P. Reis , Giuseppe Copani , Jens N. Joergensen , Bruno I. Cappellozza
Two experiments evaluated the effects of supplementing a Bacillus-based direct-fed microbial (DFM) on productive performance of mid- to late-lactating dairy cows (experiment [Exp.] 1), and on rumen fermentation traits of rumen-fistulated Holstein cows (Exp. 2). In Exp. 1, 60 mid-lactating (126 ± 11.5 DIM), primiparous (n = 14), and multiparous (n = 46) Holstein cows were blocked, within parity, by milk yield into (1) TMR (CON; n = 30) and (2) CON with Bacillus licheniformis 809 and Bacillus subtilis 810 (n = 30; BAC). Diets were offered for 12 wk, following a 3-wk covariate period. All cows received a corn silage–based TMR throughout the study. Dry matter intake and milk yield were evaluated daily, whereas milk, blood, and fecal samples were collected on wk 4, 8 and 12 for milk composition, metabolite analysis (urea and glucose), and nutrient digestibility, respectively. In Exp. 2, 16 rumen-fistulated nonlactating multiparous Holstein cows were ranked by initial BW and assigned to the treatments described in Exp. 1. The experimental period lasted 25 d and samples were taken on d 22 to 25 for in situ DM degradability, rumen ammonia, and pH. All data were analyzed with the MIXED procedure of SAS using cow as the experimental unit. In Exp. 1, DMI was reduced in BAC-fed cows, but no differences were observed on milk yield. Cows offered BAC had a grater feed efficiency versus CON (+100 g/kg feed). Bacillus spp. supplementation increased milk fat content, reduced MUN, and tended to reduce milk protein content. Mean BUN was reduced and plasma glucose was greater in cows fed BAC. No treatment effects were observed for DM or starch digestibility, but NDF digestibility was greater for BAC-fed cows. In Exp. 2, a treatment × hour interaction was observed on DM degradability, being greater at 12 h postfeeding in BAC cows versus CON. Rumen pH tended to be greater for BAC at 12, 24, and 72 h, whereas mean rumen pH and DM degradability were greater in BAC. In summary, feeding a Bacillus-based DFM to mid- to late-lactating dairy cows improved feed efficiency and increased mean glucose, while also stimulating DM and NDF digestibility.
两个试验评估了添加芽孢杆菌直接饲喂微生物(DFM)对泌乳中后期奶牛生产性能的影响(试验[试验]1),以及对瘤胃瘘管型荷斯坦奶牛瘤胃发酵特性的影响(试验2)。在试验1中,60头泌乳中期奶牛(126±11.5 DIM)、初产奶牛(n = 14)和多产奶牛(n = 46)在胎次内根据产奶量被阻断到(1)TMR (CON;n = 30)和(2)CON与地衣芽孢杆菌809和枯草芽孢杆菌810 (n = 30;BAC)。在3周的协变量期后,饲喂12周的饲粮。在整个研究过程中,所有奶牛都接受了基于玉米青贮的TMR。每天评估干物质采食量和产奶量,并在第4、8和12周采集牛奶、血液和粪便样本,分别测定乳成分、代谢物(尿素和葡萄糖)和营养物质消化率。在试验2中,选取16头瘤胃瘘非泌乳多产荷斯坦奶牛,按初始体重进行排序,并按试验1所述处理。试验期25 d,于第22 ~ 25 d取样品测定DM的原位降解率、瘤胃氨氮和ph值。以奶牛为试验单位,采用SAS的MIXED程序对所有数据进行分析。在实验1中,添加bac的奶牛DMI降低,但产奶量没有变化。饲喂BAC的奶牛饲料效率高于饲喂CON (+100 g/kg饲料)的奶牛。添加芽孢杆菌增加了乳脂含量,降低了MUN,并有降低乳蛋白含量的趋势。饲喂BAC的奶牛平均BUN降低,血浆葡萄糖升高。饲喂bac的奶牛对干物质和淀粉消化率没有影响,但NDF消化率较高。在试验2中,在DM降解率上观察到处理×小时的交互作用,在饲喂后12 h, BAC奶牛的瘤胃pH值高于对照组。在12、24和72 h, BAC奶牛的瘤胃pH值和DM降解率均高于对照组。综上所述,在泌乳中后期奶牛饲粮中添加芽孢杆菌类DFM可提高饲料效率,提高平均葡萄糖水平,同时刺激DM和NDF消化率。
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引用次数: 0
Effect of forage sorghum hybrids varying in berry size on berry processing score and in situ starch digestibility 不同浆果大小的饲用高粱杂交种对浆果加工分数和原位淀粉消化率的影响
Pub Date : 2025-07-01 DOI: 10.3168/jdsc.2025-0749
Douglas Duhatschek , Jourdan Bell , Luiz F. Ferraretto , Diego Duretto , John Goeser , Elizabeth Coons , Jason K. Smith , Sushil Paudyal , Juan M. Piñeiro
Water scarcity threatens the production of forages used to feed lactating dairy cows. Utilizing water efficient crops is key for dairy farms trying to mitigate drought and forage scarcity risks. Forage sorghum is a water-efficient crop with the potential to partially replace corn silage. However, challenges with processing forage sorghum berries have resulted in poor starch digestibility and decreased milk production of lactating dairy cows compared with those fed with corn silage. Consequently, this has prevented its adoption by dairy farmers. The objective of this study was to evaluate if a 1.70-mm berry processing score (BPS1.70) and ruminal in situ starch digestibility at 7 h (isSD7) would increase when harvesting forage sorghum with larger berry size compared with regular berry sized hybrid. Two forage sorghum hybrids (F10 and F24) were seeded in 5 plots blocked by the spans across the center irrigation pivot in a randomized complete block design. The F10 hybrid had average size berries; only 41.7% of intact berries were retained above a 3.35-mm sieve. Conversely, the F24 hybrid was genetically developed to produce a larger whole berry size and retained 90% of intact berries above a 3.35-mm sieve. With larger berries, the proportion of starch for the F24 increased almost 3 percentage units compared with F10 (26.6% and 23.9%, respectively). However, BPS1.70 and isSD7 were not different. The study suggests that although increasing berry size may increase starch content, it does not necessarily increase BPS1.70 and isSD7, at least for the berry size difference tested and harvested with kernel processors and settings commonly used to process corn.
水资源短缺威胁着奶牛饲料的生产。利用节水作物是奶牛场缓解干旱和饲料短缺风险的关键。饲用高粱是一种节水作物,具有部分替代玉米青贮的潜力。然而,与以玉米青贮饲料喂养的奶牛相比,高粱浆果饲料加工的挑战导致了淀粉消化率差和泌乳奶牛产奶量下降。因此,这阻碍了奶农采用它。本研究的目的是评估收获较大浆果尺寸的饲用高粱是否会提高1.70 mm浆果加工评分(BPS1.70)和7 h瘤胃原位淀粉消化率(isSD7)。采用完全随机区组设计,将2个饲用高粱杂交种F10和F24分在5个地块上进行播种。F10杂交种有中等大小的浆果;只有41.7%的完整浆果被保留在3.35毫米的筛子上。相反,F24杂交品种经遗传改良后,整个浆果的大小更大,在3.35毫米的筛子上保留了90%的完整浆果。在果实较大的情况下,与F10相比,F24的淀粉含量增加了近3个百分点(分别为26.6%和23.9%)。然而,BPS1.70和isSD7没有差异。研究表明,尽管增大浆果的大小可能会增加淀粉含量,但它并不一定会增加BPS1.70和isSD7,至少对于用玉米加工常用的谷粒处理机和设置测试和收获的浆果大小差异来说是这样。
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引用次数: 0
Effects of lysophosphatidylcholine on the oxidative burst response, inflammatory cytokine secretion, and Escherichia coli killing in polymorphonuclear neutrophils isolated from Holstein heifer calves 溶血磷脂酰胆碱对荷斯坦小母牛多形核中性粒细胞氧化爆发反应、炎性细胞因子分泌和大肠杆菌杀伤的影响
Pub Date : 2025-07-01 DOI: 10.3168/jdsc.2024-0646
B.N. Tate , A. Zhu , M.J.B. Felippe , D.C. Reyes , J.W. McFadden
The neonatal bovine immune system is immunonaïve at birth, making newborn calves reliant on passive immunity acquired through colostrum for protection. Calfhood illness can negatively affect health and productivity in adulthood, leading to economic losses and welfare concerns within the cattle industry. Lysophosphatidylcholine (LPC) is an immunomodulator known for enhancing bactericidal mechanisms in immune cells. However, the role of LPC in ruminants has not been thoroughly investigated. Our objective was to determine whether LPC modulates neonatal bovine neutrophil activation, cytokine release, and Escherichia coli killing. Neutrophils were isolated from the peripheral blood of preweaning Holstein heifer calves. The effects of LPC on hydrogen peroxide (H2O2) production, tumor necrosis factor-α, and IL-6 secretion, and E. coli killing were evaluated. Each experiment used 3 calves, with biological and technical replicates performed in duplicate. Data were analyzed using a mixed linear model including the fixed effects of treatment and time (when applicable) and the random effect of calf and replicate nested within treatment. In bovine neutrophils, palmitoyl-, stearoyl-, and oleoyl-LPC (i.e., LPC-16:0, -18:0, and -18:1, respectively) enhanced phorbol myristate acetate-stimulated H2O2 production. Additionally, LPC-18:0 potentiated lipopolysaccharide-induced tumor necrosis factor-α and IL-6 secretion. Although LPC-18:1 did not demonstrate this effect, LPC-18:0 was found to enhance neutrophil-mediated E. coli killing. We conclude that stearoyl-LPC enhances neutrophil bactericidal mechanisms and inflammatory responses in dairy calves. Additionally, palmitoyl- and oleoyl-LPC contribute to increased H2O2 production. These findings indicate that LPC plays an important role in modulating various neutrophil functions in neonatal bovine immunity.
新生牛的免疫系统在出生时是immunonaïve,使得新生小牛依赖于通过初乳获得的被动免疫来保护。小牛疾病会对成年期的健康和生产力产生负面影响,导致养牛业的经济损失和福利问题。溶血磷脂酰胆碱(LPC)是一种已知的免疫调节剂,可增强免疫细胞的杀菌机制。然而,LPC在反刍动物中的作用尚未得到充分的研究。我们的目的是确定LPC是否调节新生牛中性粒细胞活化、细胞因子释放和大肠杆菌杀灭。从断奶前荷斯坦小母牛犊牛外周血中分离出嗜中性粒细胞。观察LPC对过氧化氢(H2O2)生成、肿瘤坏死因子-α、IL-6分泌及大肠杆菌杀灭的影响。每个试验选用3头犊牛,生物重复和技术重复各2次。使用混合线性模型分析数据,包括处理和时间(如适用)的固定效应以及犊牛和重复巢在处理内的随机效应。在牛中性粒细胞中,棕榈酰-、硬脂酰-和油酰- lpc(分别为LPC-16:0、-18:0和-18:1)增强了肉豆蔻酸酯酚刺激的H2O2生成。此外,LPC-18:0增强了脂多糖诱导的肿瘤坏死因子-α和IL-6的分泌。虽然LPC-18:1没有表现出这种效果,但LPC-18:0被发现可以增强中性粒细胞介导的大肠杆菌杀伤。我们得出结论,硬脂酰- lpc增强了奶牛中性粒细胞杀菌机制和炎症反应。此外,棕榈酰-和油酰- lpc有助于增加H2O2的产量。这些结果表明,LPC在调节新生牛免疫中各种中性粒细胞功能中起重要作用。
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引用次数: 0
Induction of intestinal barrier dysfunction in dairy heifers: Evaluation of new serum inflammatory markers and method for quantifying intestinal hyperpermeability 奶牛肠道屏障功能障碍的诱导:新的血清炎症标志物的评价和肠道高通透性的量化方法
Pub Date : 2025-07-01 DOI: 10.3168/jdsc.2025-0740
K.E. Vagnoni, E. Lopez-Cruz, M. Carranza, D.B. Vagnoni
Experimental induction of intestinal barrier dysfunction (e.g., inflammation and hyperpermeability) has been shown to induce a systemic inflammatory response and reduce productivity in lactating dairy cows. Because numerous natural situations on-farm (e.g., ruminal acidosis, heat stress, weaning) can impair intestinal barrier function, this is an important phenomenon to study. Therefore, our objective was to induce intestinal barrier dysfunction and evaluate new serum inflammatory markers as well as a new approach to measuring intestinal hyperpermeability. This was accomplished via oral aspirin administration for 21 d in 10-mo-old Holstein and Jersey heifers. Twelve heifers (6 each, Holsteins and Jerseys) were blocked by breed and then randomly assigned (3 heifers per breed) to receive either 0 or 200 mg aspirin/kg BW per day orally. At 0600 h on d 21 of the experiment, urine and blood samples were collected from each animal. Heifers then were dosed orally with gelatin capsules containing 50 g of Co-EDTA using a balling gun. Urine samples were subsequently collected at 1, 3, 6, 8, 12, 18, 24, 30, and 36 h after dosing. Urine samples were analyzed for Co and creatinine, and serum samples were analyzed for the inflammatory markers haptoglobin (Hp), LBP, FABP2, and TNF. Modeling urinary Co:creatinine ratios using a nonlinear function yielded an excellent fit and indicated that urinary Co excretion, a measure of intestinal permeability, was not increased due to aspirin but was higher for Jersey than for Holstein heifers. Also, serum concentrations of Hp and LBP were unaffected, but serum concentrations of FABP2 and TNF were increased due to aspirin administration. Finally, analysis of covariance indicated that serum TNF concentrations were highly correlated with urinary Co excretion. These data suggest that FABP2 and TNF may be valuable additional markers for the study of intestinal barrier dysfunction.
实验表明,肠屏障功能障碍(如炎症和高渗透性)的诱导可诱导泌乳奶牛的全身炎症反应并降低生产率。由于农场的许多自然情况(如瘤胃酸中毒、热应激、断奶)会损害肠道屏障功能,这是一个重要的研究现象。因此,我们的目的是诱导肠道屏障功能障碍,评估新的血清炎症标志物以及测量肠道高通透性的新方法。这是通过10莫龄荷斯坦和泽西小母牛口服阿司匹林21 d来完成的。将12头小母牛(荷斯坦和泽西各6头)按品种隔离,然后随机分配(每个品种3头)每天口服0或200毫克/公斤体重的阿司匹林。实验第21天0600时,采集每只动物的尿液和血液样本。然后用球枪给小母牛口服含有50克Co-EDTA的明胶胶囊。随后在给药后1、3、6、8、12、18、24、30和36小时采集尿液样本。分析尿样Co和肌酐,分析血清炎症标志物触珠蛋白(Hp)、LBP、FABP2和TNF。用非线性函数模拟尿Co:肌酐比值得到了很好的拟合结果,并表明尿Co排泄量(肠道通透性的衡量指标)并未因阿司匹林而增加,但泽西奶牛的排泄量高于荷斯坦奶牛。此外,血清Hp和LBP浓度未受影响,但血清FABP2和TNF浓度因服用阿司匹林而升高。最后,协方差分析表明血清TNF浓度与尿Co排泄高度相关。这些数据表明,FABP2和TNF可能是研究肠屏障功能障碍的有价值的附加标志物。
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引用次数: 0
Relationship between cow parity and maternal parity on dairy cow lactation performance 奶牛胎次与母胎次对奶牛泌乳性能的关系
Pub Date : 2025-07-01 DOI: 10.3168/jdsc.2025-0775
D.P. Berry , K. Downing
The effect of dam parity on the subsequent performance of her female progeny is of interest when exploring the cost-benefit of alternative herd-level breeding strategies. This is especially true with the growing adoption of sexed semen where proportionally more of the heifers and younger cows in a herd may be selected as candidate dams of the next generation. There is a paucity of information on the effect of, or association between, dam parity and subsequent progeny lactation productivity in dairy cows. The objective of the cross-sectional analysis of 1,833,875 cow lactation records in the present study was to quantify the association between dam parity (1 to 10+) and subsequent progeny lactation yield, milk fat and protein concentration, and SCC. Also of interest were model cow parity effects for these traits so as to inform cost-benefit analyses of improving cow lifespan. All associations were estimated using linear mixed models with cow included as a random effect while also facilitating heterogeneous dam and cow parity variance components. Mean yield increased with cow parity up to parity 5 (30% to 33% higher than yields from first parity cows) after which it declined; parity 10+ cows still produced, on average, 19% to 21% more milk than first parity cows although mean lactation SCC increased with advancing parity in multiparous cows. The mean 305-d lactation milk yield of cows increased as the parity number of their dams increased, peaking at parity 7 (19.8 kg more milk than progeny from first lactation dams) before declining. In contrast, peak fat and protein yields occurred at younger dam parities. Although the mean SCS of cows was greatest in progeny from parity 2 dams, mean lactation SCS declined thereafter as dam parity number increased. In conclusion, although dam parity was associated with progeny lactation performance, the biological extent of this association was almost negligible.
在探索其他群体水平繁殖策略的成本效益时,大坝胎次对其雌性后代后续表现的影响是令人感兴趣的。这在越来越多地采用有性精子的情况下尤其如此,在这种情况下,按比例,一群母牛中有更多的小母牛和年轻的母牛可能被选为下一代的候选水坝。在奶牛中,关于胎次与后代泌乳率的影响或关联的信息缺乏。本研究对1,833,875头奶牛的泌乳记录进行了横断面分析,目的是量化胎次(1 ~ 10+)与随后的后代泌乳量、乳脂和蛋白质浓度以及SCC之间的关系。同样令人感兴趣的是模型牛胎次对这些性状的影响,以便为提高奶牛寿命的成本效益分析提供信息。所有关联都使用线性混合模型进行估计,其中包括奶牛作为随机效应,同时也促进了异质性大坝和奶牛宇称方差成分。与牛平价意味着产量增加到平价5(30%对33%高于产量从第一平价牛)后拒绝了;胎次10胎以上的奶牛产奶量仍比第一次胎次的奶牛平均多出19%至21%,尽管多产奶牛的平均泌乳SCC随着胎次的增加而增加。奶牛305 d平均泌乳产奶量随着胎次的增加而增加,在胎次7时达到峰值(比第一个泌乳胎多产奶19.8 kg),然后下降。相反,脂肪和蛋白质产量的峰值出现在较年轻的坝仔。虽然胎次2次的奶牛的平均SCS最高,但随着胎次数的增加,奶牛的平均泌乳SCS逐渐下降。综上所述,虽然胎次与后代的泌乳性能有关,但这种联系的生物学程度几乎可以忽略不计。
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引用次数: 0
Evaluating bacterial growth in raw, frozen, and heat-treated colostrum inoculated with fecal Escherichia coli 评估接种粪便大肠杆菌的生的、冷冻的和热处理的初乳中的细菌生长情况
Pub Date : 2025-07-01 DOI: 10.3168/jdsc.2025-0756
A.M. McKane, T.A. Westhoff, S. Klaessig, C. Altier, K.E. Bell, P.D. Pavinski Bitar, S. Mann
Providing newborn calves with sufficient high-quality colostrum is a critical calf-management strategy to support health, survival, and future productivity. Unfortunately, colostrum may also serve as a fomite for disease when contaminated with bacteria at harvest, during storage, or during reheating before feeding. Thermal processing, including heat treatment (HT; 60°C for 60 min) and freezing (FR; −20°C), are common strategies to manage bacterial contamination. Although both strategies maintain IgG concentrations, they destroy colostral immune cells, and HT is known to decrease the concentration and activity of certain bioactive factors such as complement and IgA. We hypothesized that HT and FR would influence the inherent antibacterial properties of bovine colostrum compared with unprocessed, refrigerated colostrum (RW). Our objective was thus to compare growth of Escherichia coli in RW, HT, and FR bovine colostrum. Sterile colostrum samples were collected from Holstein cows (n = 11) on a commercial dairy in New York State and divided into 4 aliquots. One aliquot was submitted for bacterial culture to exclude samples with any bacterial growth (n = 4). The remaining 7 samples were used for the experiment. Aliquots were either RW (4°C for 20 h; n = 7), HT (60°C for 60 min, then 4°C for 19 h; n = 7), or FR (−20°C for 20 h; n = 7). Immediately before inoculation of samples for a bacterial kinetics assay, a dried bovine colostrum-based replacer (CR; Ultra Start 150, Sav-A-Calf, Chilton, WI) was prepared from a single bag according to package instructions as a nutrient-rich control (n = 7). The prepared CR aliquots underwent bacterial culture to ascertain the absence of bacterial growth before inoculation. To simulate contamination that may occur on-farm, all samples were inoculated with 104 cfu/mL E. coli (WM1; 060913 P0lA, isolated from bovine feces) and tested in a bacterial kinetics assay at 37°C. At 0, 2, 4, 6, 8, and 24 h, growth of E. coli was quantified on MacConkey agar plates inoculated at 37°C. Relative to colostrum replacer, growth of E. coli was lower in RW and FR colostrum from 4 to 24 h and lower in HT colostrum at 6 and 8 h. These results demonstrate inherent microbial growth-inhibiting activity in RW colostrum and suggest that FR better preserved this activity than HT. Our study contributes to understanding the effect of thermal processing on antimicrobial properties of colostrum with the goal of helping to inform colostrum-management strategies for dairy producers. Specifically, our data emphasizes the need to practice hygiene during all steps of the colostrum harvest and storage process, including after completion of HT, because this renders colostrum more susceptible to bacterial growth. Similarly, the cooling and heating steps of stored colostrum should be rapid to minimize growth of E. coli.
为新生牛犊提供足够的高质量初乳是一项重要的小牛管理策略,以支持其健康、生存和未来的生产力。不幸的是,当初乳在收获、储存或喂养前加热过程中被细菌污染时,也可能成为疾病的病原体。热处理,包括热处理(HT;60°C 60分钟)和冷冻(FR;−20°C),是控制细菌污染的常用策略。虽然这两种策略都维持IgG浓度,但它们破坏了初乳免疫细胞,并且已知HT会降低某些生物活性因子(如补体和IgA)的浓度和活性。我们假设HT和FR会影响牛初乳与未加工的冷藏初乳(RW)的固有抗菌性能。因此,我们的目的是比较大肠杆菌在RW、HT和FR牛初乳中的生长情况。从纽约州一家商业奶牛场的荷斯坦奶牛(n = 11)中采集无菌初乳样品,分成4份。其中一份样品进行细菌培养,以排除任何细菌生长的样品(n = 4)。剩余7份样品用于实验。等分分别为RW(4°C) 20 h;n = 7),高温(60°C 60 min,然后4°C 19 h;n = 7),或FR(- 20°C, 20 h;N = 7)。在接种样品进行细菌动力学试验之前,立即使用基于干牛初乳的替代品(CR;Ultra Start 150, Sav-A-Calf, Chilton, WI)根据包装说明从一个袋子中制备,作为营养丰富的对照(n = 7)。制备的CR等分液在接种前进行细菌培养以确定没有细菌生长。为了模拟农场可能发生的污染,所有样品接种104 cfu/mL大肠杆菌(WM1;060913 P0lA,从牛粪便中分离),并在37°C的细菌动力学试验中进行测试。在0、2、4、6、8和24 h,在37℃接种的MacConkey琼脂板上定量大肠杆菌的生长。与替代初乳相比,RW和FR初乳在4 ~ 24 h期间的大肠杆菌生长较低,HT初乳在6和8 h时的生长较低。这些结果表明,RW初乳具有内在的微生物生长抑制活性,FR比HT更能保持这种活性。我们的研究有助于了解热加工对初乳抗菌特性的影响,目的是帮助乳品生产商提供初乳管理策略。具体来说,我们的数据强调在初乳收获和储存过程的所有步骤中,包括完成高温处理后,都需要实践卫生,因为这使得初乳更容易受到细菌生长的影响。同样,储存初乳的冷却和加热步骤应迅速,以尽量减少大肠杆菌的生长。
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引用次数: 0
Validation of a rapid on-farm culture system for group classification of clinical mastitis-causing pathogens 用于临床乳腺炎致病菌分类的快速农场培养系统的验证
Pub Date : 2025-07-01 DOI: 10.3168/jdsc.2024-0738
Fernando J. Guardado Servellon , David L. Renaud , Bruno Joaquin Paredes Osorio , Kelsey L. Spence , Trevor J. DeVries , Rita Couto Serrenho
The objective of this diagnostic accuracy study was to compare the use of a rapid tube test system (MastDecide [MD]; Quidee GmbH, Homberg, Germany) to aerobic milk culture for group classification of mastitis pathogens in dairy cattle. A total of 204 milk samples from cows with clinical mastitis (CM) were collected across 60 dairy herds in Ontario, Canada. Samples were collected by dairy producers and their staff and transported refrigerated to the University of Guelph Animal Health Laboratory (Guelph, ON, Canada) on the day of collection. Each sample was tested via MD (gram-positive, gram-negative, or “no growth”) and via aerobic milk culture followed by MALDI-TOF (reference method [RM]). The MD and RM results were interpreted at 14 and 24 h after incubation, respectively. An additional assessment was performed regarding the accuracy of intramammary antibiotic treatment decisions (gram-positive: to treat; gram-negative and “no growth” samples: not to treat). Test performance characteristics (overall accuracy, sensitivity [Se], specificity [Sp], and positive [PPV] and negative predictive value [NPV]) as well as Cohen's kappa coefficient (κ) were calculated. The RM results were classified as gram-positive (n = 107, 53%), gram-negative (n = 20, 10%), “no growth” (n = 51, 25%), other pathogens (n = 21, 10%), and mixed growth (gram-positive and gram-negative; n = 5, 3%). The Se of the MD test was 58% (95% CI: 47%–67%), 40% (19%–64%), and 61% (50%–72%) for gram-positive, gram-negative, and “no growth,” respectively. The Sp of the MD test was 73% (63%–81%), 84% (78%–89%), and 75% (66%–82%) for gram-positive, gram-negative, and “no growth,” respectively. The PPV of the MD test was 68% (57%–78%), 22% (10%–39%), and 62% (50%–73%) for gram-positive, gram-negative, and “no growth,” respectively; and the NPV was 63% (54%–72%), 93% (87%–96%), and 74% (65%–82%) for gram-positive, gram-negative, and “no growth,” respectively. The overall test accuracy for Gram stain classification analysis was 57%, with a κ of 0.30. Regarding intramammary antibiotic treatment, the MD test yielded a correct decision relative to the RM for 65% of the cases. Using this on-farm test as a primary guide in CM treatment decisions should be approached with caution, as the risk of false negatives may affect the ability to effectively treat CM cases.
本诊断准确性研究的目的是比较快速试管测试系统(MastDecide [MD];Quidee GmbH, Homberg, Germany)采用有氧乳培养对奶牛乳腺炎病原体进行分组分类。在加拿大安大略省的60个奶牛群中收集了204个临床乳腺炎(CM)奶牛的牛奶样本。样品由乳制品生产商及其工作人员收集,并在收集当天冷藏运送到圭尔夫大学动物卫生实验室(加拿大圭尔夫,ON, Canada)。每个样品通过MD(革兰氏阳性、革兰氏阴性或“无生长”)和有氧乳培养进行检测,然后进行MALDI-TOF(参考方法[RM])。MD和RM结果分别在孵育后14和24 h进行解释。对乳腺内抗生素治疗决策的准确性进行了额外评估(革兰氏阳性:治疗;革兰氏阴性和“无生长”样本:不治疗)。计算试验性能特征(总体准确性、灵敏度[Se]、特异性[Sp]、阳性预测值[PPV]和阴性预测值[NPV])以及科恩kappa系数(κ)。RM结果分为革兰氏阳性(n = 107, 53%)、革兰氏阴性(n = 20, 10%)、“无生长”(n = 51, 25%)、其他病原体(n = 21, 10%)和混合生长(革兰氏阳性和革兰氏阴性;N = 5,3 %)。革兰氏阳性、革兰氏阴性和“无生长”的MD检测Se分别为58% (95% CI: 47%-67%)、40%(19%-64%)和61%(50%-72%)。对于革兰氏阳性、革兰氏阴性和“无生长”,MD试验的Sp分别为73%(63% ~ 81%)、84%(78% ~ 89%)和75%(66% ~ 82%)。对于革兰氏阳性、革兰氏阴性和“无生长”,MD试验的PPV分别为68%(57%-78%)、22%(10%-39%)和62% (50%-73%);革兰氏阳性、革兰氏阴性和“无生长”的净现值分别为63%(54% ~ 72%)、93%(87% ~ 96%)和74%(65% ~ 82%)。革兰氏染色分类分析的总体检测准确率为57%,κ为0.30。关于乳腺内抗生素治疗,相对于RM, MD测试对65%的病例产生了正确的决定。使用这种农场测试作为CM治疗决策的主要指导应谨慎对待,因为假阴性的风险可能影响有效治疗CM病例的能力。
{"title":"Validation of a rapid on-farm culture system for group classification of clinical mastitis-causing pathogens","authors":"Fernando J. Guardado Servellon ,&nbsp;David L. Renaud ,&nbsp;Bruno Joaquin Paredes Osorio ,&nbsp;Kelsey L. Spence ,&nbsp;Trevor J. DeVries ,&nbsp;Rita Couto Serrenho","doi":"10.3168/jdsc.2024-0738","DOIUrl":"10.3168/jdsc.2024-0738","url":null,"abstract":"<div><div>The objective of this diagnostic accuracy study was to compare the use of a rapid tube test system (MastDecide [MD]; Quidee GmbH, Homberg, Germany) to aerobic milk culture for group classification of mastitis pathogens in dairy cattle. A total of 204 milk samples from cows with clinical mastitis (CM) were collected across 60 dairy herds in Ontario, Canada. Samples were collected by dairy producers and their staff and transported refrigerated to the University of Guelph Animal Health Laboratory (Guelph, ON, Canada) on the day of collection. Each sample was tested via MD (gram-positive, gram-negative, or “no growth”) and via aerobic milk culture followed by MALDI-TOF (reference method [RM]). The MD and RM results were interpreted at 14 and 24 h after incubation, respectively. An additional assessment was performed regarding the accuracy of intramammary antibiotic treatment decisions (gram-positive: to treat; gram-negative and “no growth” samples: not to treat). Test performance characteristics (overall accuracy, sensitivity [Se], specificity [Sp], and positive [PPV] and negative predictive value [NPV]) as well as Cohen's kappa coefficient (κ) were calculated. The RM results were classified as gram-positive (n = 107, 53%), gram-negative (n = 20, 10%), “no growth” (n = 51, 25%), other pathogens (n = 21, 10%), and mixed growth (gram-positive and gram-negative; n = 5, 3%). The Se of the MD test was 58% (95% CI: 47%–67%), 40% (19%–64%), and 61% (50%–72%) for gram-positive, gram-negative, and “no growth,” respectively. The Sp of the MD test was 73% (63%–81%), 84% (78%–89%), and 75% (66%–82%) for gram-positive, gram-negative, and “no growth,” respectively. The PPV of the MD test was 68% (57%–78%), 22% (10%–39%), and 62% (50%–73%) for gram-positive, gram-negative, and “no growth,” respectively; and the NPV was 63% (54%–72%), 93% (87%–96%), and 74% (65%–82%) for gram-positive, gram-negative, and “no growth,” respectively. The overall test accuracy for Gram stain classification analysis was 57%, with a κ of 0.30. Regarding intramammary antibiotic treatment, the MD test yielded a correct decision relative to the RM for 65% of the cases. Using this on-farm test as a primary guide in CM treatment decisions should be approached with caution, as the risk of false negatives may affect the ability to effectively treat CM cases.</div></div>","PeriodicalId":94061,"journal":{"name":"JDS communications","volume":"6 4","pages":"Pages 562-566"},"PeriodicalIF":0.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144517986","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Skimming and storage factors affect the detection of heat shock protein 70 in raw bovine milk 脱脂和贮存因素影响生牛乳中热休克蛋白70的检测
Pub Date : 2025-07-01 DOI: 10.3168/jdsc.2025-0759
M.R.H. Rakib , V. Messina , J.I. Gargiulo , N.A. Lyons , I.N. Pathirana , P.C. Thomson , S.C. Garcia
Heat shock proteins (HSP) play a vital role in maintaining cellular function under elevated temperatures. Among these, heat shock protein 70 (HSP70) has recently been detected in bovine milk using competitive ELISA, highlighting its potential as a noninvasive marker of cellular stress and a tool for detecting heat stress (HS) in dairy cows. However, optimal management and storage conditions of milk samples for detection of HSP70 are yet to be determined. This study aimed to evaluate how different management and storage practices affect detection of HSP70 in milk samples. Samples were skimmed by centrifugation either before storage (SBS) or after storage (SAS), and then stored under different temperatures: (a) room temperature (T1, ∼22°C), (b) refrigerated (T2, 4°C), (c) frozen (T3, −20°C), and (d) refrigerated at 4°C with a preservative (bronopol; T4). Samples were analyzed every 2 d until d 15 (d 1, d 3, …, d 15) using a competitive ELISA system (sensitivity range 31.25–2,000 ng/mL) to detect HSP70 levels. Result showed that SBS samples had the smallest reduction in HSP70 levels on d 3 when stored at T2 or T3, with decreases of 4.4% and 7.9%, respectively. In contrast, SAS samples exhibited greater reductions (44.2% at T2 and 53.9% at T3). Samples stored at room temperature (T1) showed the greatest degradation, especially in SAS samples, whereas the use of a preservative (T4) did not consistently maintain HSP70 levels in either SBS or SAS samples. The HSP70 detection showed lower variability during early storage (d 1–3) across all treatments, with SBS samples demonstrating greater stability than SAS; however, variability significantly increased by d 9 in both sample types. An apparent increase in HSP70 levels was observed from d 11 to 15 in both SBS and SAS, suggesting the influence of protein stability and interactions affecting ELISA reactivity, where further research is required. Overall, samples stored as skim milk until d 3 at 4°C or −20°C preserved HSP70 levels more effectively than whole milk, with room temperature storage being the least favorable. This pioneering study explored how sample storage and management affect HSP70 detection, providing practical guidelines for farmers and processors to preserve milk sample integrity for assessment of HSP70 as indicator of cellular stress and, potentially, HS.
热休克蛋白在高温下维持细胞功能方面起着至关重要的作用。其中,热休克蛋白70 (HSP70)最近通过竞争性ELISA在牛奶中检测到,突出了它作为一种无创细胞应激标记物和检测奶牛热应激(HS)的工具的潜力。然而,检测HSP70的牛奶样品的最佳管理和储存条件尚未确定。本研究旨在评估不同的管理和储存方式对牛奶样品HSP70检测的影响。样品在储存前(SBS)或储存后(SAS)通过离心脱脂,然后在不同的温度下储存:(a)室温(T1, ~ 22°C), (b)冷藏(T2, 4°C), (C)冷冻(T3, - 20°C), (d)在4°C冷藏,并添加防腐剂(bronopol;T4)。使用竞争性ELISA系统(灵敏度范围31.25 ~ 2000 ng/mL)检测HSP70水平,每2天分析一次样品,直到第15天(第1、3、…、15天)。结果表明,SBS样品在T2和T3贮藏时,第3天HSP70水平下降幅度最小,分别为4.4%和7.9%。相反,SAS样品表现出更大的减少(在T2时为44.2%,在T3时为53.9%)。在室温(T1)下保存的样品表现出最大的降解,特别是在SAS样品中,而使用防腐剂(T4)并不能始终保持SBS或SAS样品中的HSP70水平。在所有处理中,HSP70检测在贮藏早期(1-3天)的变异性较低,SBS样品比SAS样品表现出更大的稳定性;然而,在两种样本类型中,变异性显著增加了9。从第11天到第15天,SBS和SAS的HSP70水平明显升高,这表明蛋白质稳定性和相互作用影响了ELISA反应性,这需要进一步的研究。总体而言,在4°C或- 20°C下以脱脂牛奶形式保存至3 d的样品比全脂牛奶更有效地保存了HSP70水平,而室温保存效果最差。这项开创性的研究探讨了样品储存和管理如何影响HSP70检测,为农民和加工商提供实用指南,以保持牛奶样品的完整性,以评估HSP70作为细胞应激和潜在HS的指标。
{"title":"Skimming and storage factors affect the detection of heat shock protein 70 in raw bovine milk","authors":"M.R.H. Rakib ,&nbsp;V. Messina ,&nbsp;J.I. Gargiulo ,&nbsp;N.A. Lyons ,&nbsp;I.N. Pathirana ,&nbsp;P.C. Thomson ,&nbsp;S.C. Garcia","doi":"10.3168/jdsc.2025-0759","DOIUrl":"10.3168/jdsc.2025-0759","url":null,"abstract":"<div><div>Heat shock proteins (HSP) play a vital role in maintaining cellular function under elevated temperatures. Among these, heat shock protein 70 (HSP70) has recently been detected in bovine milk using competitive ELISA, highlighting its potential as a noninvasive marker of cellular stress and a tool for detecting heat stress (HS) in dairy cows. However, optimal management and storage conditions of milk samples for detection of HSP70 are yet to be determined. This study aimed to evaluate how different management and storage practices affect detection of HSP70 in milk samples. Samples were skimmed by centrifugation either before storage (SBS) or after storage (SAS), and then stored under different temperatures: (a) room temperature (T<sub>1</sub>, ∼22°C), (b) refrigerated (T<sub>2</sub>, 4°C), (c) frozen (T<sub>3</sub>, −20°C), and (d) refrigerated at 4°C with a preservative (bronopol; T<sub>4</sub>). Samples were analyzed every 2 d until d 15 (d 1, d 3, …, d 15) using a competitive ELISA system (sensitivity range 31.25–2,000 ng/mL) to detect HSP70 levels. Result showed that SBS samples had the smallest reduction in HSP70 levels on d 3 when stored at T<sub>2</sub> or T<sub>3</sub>, with decreases of 4.4% and 7.9%, respectively. In contrast, SAS samples exhibited greater reductions (44.2% at T<sub>2</sub> and 53.9% at T<sub>3</sub>). Samples stored at room temperature (T<sub>1</sub>) showed the greatest degradation, especially in SAS samples, whereas the use of a preservative (T<sub>4</sub>) did not consistently maintain HSP70 levels in either SBS or SAS samples. The HSP70 detection showed lower variability during early storage (d 1–3) across all treatments, with SBS samples demonstrating greater stability than SAS; however, variability significantly increased by d 9 in both sample types. An apparent increase in HSP70 levels was observed from d 11 to 15 in both SBS and SAS, suggesting the influence of protein stability and interactions affecting ELISA reactivity, where further research is required. Overall, samples stored as skim milk until d 3 at 4°C or −20°C preserved HSP70 levels more effectively than whole milk, with room temperature storage being the least favorable. This pioneering study explored how sample storage and management affect HSP70 detection, providing practical guidelines for farmers and processors to preserve milk sample integrity for assessment of HSP70 as indicator of cellular stress and, potentially, HS.</div></div>","PeriodicalId":94061,"journal":{"name":"JDS communications","volume":"6 4","pages":"Pages 578-582"},"PeriodicalIF":0.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144517905","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Effects of large follicle numbers at the onset of short-term timed artificial insemination protocol in lactating dairy cows 短期定时人工授精开始时大卵泡数对泌乳奶牛的影响
Pub Date : 2025-07-01 DOI: 10.3168/jdsc.2024-0682
Kazuhiro Bandai , Ena Chiba , Toru Ogata , Yoshiyuki Tsuchiya , Tadaharu Ajito , Ryotaro Miura
This study aimed to compare the fertility of lactating Holstein cows with only 1 large follicle (LF, diameter ≥10 mm) with those with ≥2 LF and corpora lutea at the start of short-term ovulation synchronization and timed artificial insemination (AI). Cows with a corpus luteum ≥20 mm in diameter and either 1 LF (1F) or ≥2 LF (2F) verified via ovarian ultrasonography were included in the study. Cows were randomly categorized into groups receiving either estradiol benzoate (EB) or GnRH as ovulation induction agents. Cows received a luteolytic dose of PGF, followed by administration of EB 24 h later or GnRH 56 h later. Timed AI was performed 24 to 28 h after EB administration or 16 to 20 h after GnRH administration. We analyzed data from 605 AI in 554 cows (1F-EB: n = 193, 2F-EB: n = 100, 1F-GnRH: n = 213, 2F-GnRH: n = 99). Logistic regression analysis revealed a significant interaction between the number of LF and the ovulation induction agent. Pregnancy per AI was significantly lower in the 1F-GnRH group than in the other 3 groups (1F-EB: 40.9%, 2F-EB: 44.0%, 1F-GnRH: 28.2%, 2F-GnRH: 44.4%). In conclusion, the fertility of cows with 1F was lower than that of cows with 2F when GnRH was administered as the ovulation induction agent in the ovulation synchronization and timed AI protocol. However, the fertility of 1F cows receiving EB was comparable to that of 2F cows receiving either GnRH or EB.
本研究旨在比较短期同步排卵和定时人工授精(AI)开始时,只有1个大卵泡(LF,直径≥10 mm)的泌乳荷斯坦奶牛与有≥2个大卵泡和黄体的泌乳荷斯坦奶牛的生育能力。黄体直径≥20 mm且卵巢超声检查证实为1 LF (1F)或≥2 LF (2F)的奶牛纳入研究。奶牛随机分为两组,分别给予苯甲酸雌二醇(EB)和GnRH促排卵剂。母牛接受促黄体溶解剂量的PGF2α, 24 h后给予EB, 56 h后给予GnRH。在EB给药后24 ~ 28 h或GnRH给药后16 ~ 20 h进行定时人工智能。我们分析了554头奶牛605只AI的数据(1F-EB: n = 193, 2F-EB: n = 100, 1F-GnRH: n = 213, 2F-GnRH: n = 99)。Logistic回归分析显示LF数量与促排卵剂之间存在显著的交互作用。1F-GnRH组每AI妊娠率明显低于其他3组(1F-EB: 40.9%, 2F-EB: 44.0%, 1F-GnRH: 28.2%, 2F-GnRH: 44.4%)。综上所述,在同步排卵和定时人工授精方案中,GnRH作为促排卵剂,1F奶牛的生育能力低于2F奶牛。然而,接受EB的1F奶牛的生育能力与接受GnRH或EB的2F奶牛相当。
{"title":"Effects of large follicle numbers at the onset of short-term timed artificial insemination protocol in lactating dairy cows","authors":"Kazuhiro Bandai ,&nbsp;Ena Chiba ,&nbsp;Toru Ogata ,&nbsp;Yoshiyuki Tsuchiya ,&nbsp;Tadaharu Ajito ,&nbsp;Ryotaro Miura","doi":"10.3168/jdsc.2024-0682","DOIUrl":"10.3168/jdsc.2024-0682","url":null,"abstract":"<div><div>This study aimed to compare the fertility of lactating Holstein cows with only 1 large follicle (LF, diameter ≥10 mm) with those with ≥2 LF and corpora lutea at the start of short-term ovulation synchronization and timed artificial insemination (AI). Cows with a corpus luteum ≥20 mm in diameter and either 1 LF (1F) or ≥2 LF (2F) verified via ovarian ultrasonography were included in the study. Cows were randomly categorized into groups receiving either estradiol benzoate (EB) or GnRH as ovulation induction agents. Cows received a luteolytic dose of PGF<sub>2α</sub>, followed by administration of EB 24 h later or GnRH 56 h later. Timed AI was performed 24 to 28 h after EB administration or 16 to 20 h after GnRH administration. We analyzed data from 605 AI in 554 cows (1F-EB: n = 193, 2F-EB: n = 100, 1F-GnRH: n = 213, 2F-GnRH: n = 99). Logistic regression analysis revealed a significant interaction between the number of LF and the ovulation induction agent. Pregnancy per AI was significantly lower in the 1F-GnRH group than in the other 3 groups (1F-EB: 40.9%, 2F-EB: 44.0%, 1F-GnRH: 28.2%, 2F-GnRH: 44.4%). In conclusion, the fertility of cows with 1F was lower than that of cows with 2F when GnRH was administered as the ovulation induction agent in the ovulation synchronization and timed AI protocol. However, the fertility of 1F cows receiving EB was comparable to that of 2F cows receiving either GnRH or EB.</div></div>","PeriodicalId":94061,"journal":{"name":"JDS communications","volume":"6 4","pages":"Pages 589-592"},"PeriodicalIF":0.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144517957","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Factors affecting follicular emergence at 10 days postpartum in lactating dairy cows 影响泌乳奶牛产后10天卵泡出现的因素
Pub Date : 2025-07-01 DOI: 10.3168/jdsc.2025-0744
Hiromi Kusaka , Takeshi Yamazaki , Minoru Sakaguchi
Several factors influence postpartum ovarian activity in dairy cows. Our previous research confirmed a significant seasonal effect on the occurrence of early ovulation until 26 d postpartum, yet the relationships with earlier follicular development were not examined. This retrospective study aimed to describe the association between follicular development at 10 d postpartum and the presence of early ovulation in 536 lactations. Furthermore, the factors affecting the emergence of ≥15 mm (LL-size) of follicles at 10 d postpartum were assessed, including calving season, parity of dams, BW loss, period of years, number of calves, and uterine abnormality. Approximately 12% of cows lacked ≥10.0 mm follicles at 10 d postpartum and had a lower frequency of early ovulation (32.3%). Early ovulation was observed more in cows with at least one LL-size but no 10.0 to 14.9 mm follicles (69.7%). The odds ratio of LL-size follicle presence in the cows with high uterine abnormalities was significantly lower than in the cows with low uterine abnormalities (odds ratio: 0.20, 54.0% vs. 83.5%). Fewer <10.0 mm follicles were detected in the former than the latter. The cows that calved in summer had a higher frequency of LL-size follicles than those that calved in winter and spring. The effects of BW loss and dam parity on the presence of LL-size follicles approached significance. In conclusion, calving season, BW loss, and dam parity influenced the likelihood of LL-size follicle emergence. In particular, an abnormal uterine status 10 d after calving inhibited early follicular development, which might affect early ovulation.
影响奶牛产后卵巢活动的几个因素。我们之前的研究证实,在产后26天之前,季节性因素对早期排卵的发生有显著影响,但与早期卵泡发育的关系尚未得到检验。本回顾性研究旨在描述536例哺乳期患者产后10天卵泡发育与早期排卵之间的关系。此外,评估影响产后10 d出现≥15mm (LL-size)卵泡的因素,包括产犊季节、胎次、体重损失、产龄、产仔数和子宫异常。约12%的奶牛在产后10天卵泡缺失≥10.0 mm,早期排卵频率较低(32.3%)。至少1个l -大小但没有10.0 ~ 14.9 mm卵泡的奶牛早期排卵较多(69.7%)。子宫高度异常奶牛l -大小卵泡存在的比值比显著低于子宫轻度异常奶牛(比值比:0.20,54.0% vs. 83.5%)。前者比后者少检出10.0 mm的卵泡。在夏季产犊的奶牛比在冬季和春季产犊的奶牛有更高的l -大小卵泡的频率。体重损失和胎次对l -大小卵泡存在的影响接近显著。综上所述,产犊季节、体重损失和胎次影响卵泡出现的可能性。特别是产蛋后10天子宫状态异常会抑制早期卵泡发育,影响早期排卵。
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