餵飼新穎益生菌組合對荷蘭乳牛和仔牛生產表現和健康的影響

Abstract

在畜牧業中，益生菌的使用已有多年的歷史，隨著精準醫療的快速發展，次世代益生菌的應用潛力日益受到重視，開發適用於乳牛的高效益生菌產品已成為一項重要的研究議題，然而，目前對乳牛新一代益生菌的研究仍然相當有限且菌株多半非源自乳牛，而且多數研究僅進行了短期的評估，有鑑於此，本研究旨在探討一種由三種菌株組成的新型益生菌配方，包括B. longum APL30、L. salivarius K108 和 S. cerevisiae T15，對於荷蘭乳牛和仔牛的健康狀況和生產性能的影響，這項牛隻試驗在台北和台南兩個不同的牧場進行。 在台北牧場的試驗中，我們餵飼18頭泌乳牛益生菌180天，並在試驗第0天 (T0)、試驗第60天 (T1) 和試驗第180天 (T2)，收集了血液、糞便和瘤胃液等樣品進行分析。在台南牧場試驗中，50頭懷孕乳牛被隨機分為對照組 (C組) 和益生菌組 (P組)，試驗總共進行240天直到母牛分娩後2個月，以評估益生菌長期補充對牛隻健康的影響，整個試驗過程會收集每個月疾病用藥紀錄與乳量與乳品質報告，並在試驗第0天 (S0)、試驗第60天 (S1)、乾乳期 (S2)、分娩期 (S3) 和產後2個月 (S4) 收集血液、糞便和瘤胃液等樣品進行分析，同時，牠們所產下的仔牛也進一步納入仔牛試驗直到2個月齡。仔牛共83頭分為3個組別，包括了母牛與仔牛皆未使用益生菌的C組，母牛未使用仔牛使用益生菌的N組，與母牛和仔牛皆使用益生菌的P組，並會在仔牛出生0天、30天和60天測量體重，並收集血液和糞便。 結果在台南牧場的試驗中，P組牛隻的疾病和乳房炎發生率顯著低於對照組，尤其是初產牛表現更為突出; 牛乳中體細胞數 (somatic cell count, SCC) 方面，台北牧場大多數牛隻維持在理想範圍，且經產牛與前一胎次SCC相比也具有較低數量，台南牧場益生菌組牛隻SCC值亦顯著低於對照組，在泌乳初期 (泌乳天數 0-150天)，補充益生菌後牛隻體態評分 (body condition score) 也有所提升，另外，在分娩後餵飼益生菌可提升牛隻乳量，乳蛋白率和酪蛋白率的含量。透過瘤胃發酵代謝和菌相方面進行了探討本益生菌可能的作用途徑，在瘤胃總揮發性脂肪酸 (volatile fatty acids, VFA) 含量方面，台北牧場T1階段顯著升高，而台南牧場S4階段則顯著降低，各類VFA的比例變化也不太一致，但兩牧場觀察到異丁酸以及微生物體蛋白 (microbial crude protein, MCP) 含量升高，其增加可能是因益生菌能夠促進瘤胃內胺基酸的有效代謝和利用。在菌相方面，補充益生菌顯著提升了瘤胃內微生物的多樣性。我們觀察到益生菌能夠增加健康泌乳牛生物指標菌種Ruminococcus與Bifidobacterium的相對豐度， 此外，我們還發現補充益生菌能夠增加與MCP合成呈正相關的Breznakia pachnodae和Sodaliphilus pleomorphus的含量，功能預測分析顯示補充益生菌的牛隻瘤胃菌相在胺基酸和碳水化合物代謝路徑方面有顯著增加，這表明益生菌能夠藉由影響瘤胃微生物群與其代謝活動，促進牛隻對營養物質的消化和吸收，從而提高健康與生產性能。 在仔牛方面，在補充益生菌後仔牛的腹瀉發生率也明顯降低，菌相的部分發現到與糞便評分呈負相關的Pseudoramibacter、Pseudoramibacter porci 與Solibacillus isronensis B3W22增加，並在基因功能性的預測觀察到胺基酸代謝路徑與訊息傳遞路徑的增加，說明益生菌可能藉由調節腸道菌相，改善消化吸收和免疫功能，從而提高仔牛的疾病抵抗力。 綜上所述，本研究證實了新穎益生菌組合在不同牧場環境下對荷蘭母牛和仔牛的健康狀況及生產性能均存在正面影響，並具有減少抗生素使用、增加經濟效益和提升乳牛福祉在畜牧業中的實際應用潛力。

Probiotics have been used in ruminant production for many years. With the rapid development of precision medicine, the application potential of next-generation probiotics is increasingly gaining attention. The development of highly effective probiotic products for dairy cows has become an important research topic. However, research on next-generation probiotics for dairy cows is still quite limited. It is worth noting that many previous dairy cow probiotic trials used strains not derived from dairy cows, and most studies only conducted short-term evaluations. In view of this, the present study aims to investigate the effects of a new probiotic formula composed of three strains, including Bifidobacterium longum subsp. longum APL30, Ligilactobacillus salivarius K108, and Saccharomyces cerevisiae T15 on the health status and production performance of Holstein cows and calves. This cattle trial was conducted on two different farms in Taipei and Tainan. In the trial at the Taipei farm, we fed 18 lactating cows probiotics for 180 days and collected samples such as blood, feces, and rumen fluid for analysis on day 0 (T0), day 60 (T1), and day 180 (T2) of the trial. In the trial at the Tainan farm, 50 pregnant dairy cows were randomly divided into a control group (C group) and a probiotic group (P group). The trial lasted a total of 240 days until the cows calved and continued for 2 months post-calving to assess the long-term effects of probiotic supplementation on cow health. Throughout the trial, records of disease medication, milk production, and milk quality reports were collected monthly. Blood, feces, and rumen fluid samples were collected for analysis on day 0 (S0), day 60 (S1), dry period (S2), calving period (S3), and 2 months post-calving (S4). Additionally, the calves produced were further included in a calf trial until they reached 2 months of age. A total of 83 calves were divided into 3 groups: the control group (C group) where neither the cows nor calves received probiotics, the N group where the cows did not receive probiotics but the calves did, and the P group where both the cows and calves received probiotics. We measured the calves' weights at birth, 30 days, and 60 days, and collected blood and feces samples. For cows, the trial conducted at the Tainan farm showed that the incidence of diseases and mastitis in the P group was significantly lower than in the control group, especially in primiparous cows. Regarding somatic cell counts (SCC), most cows at the Taipei farm maintained an ideal range. At the Tainan farm, the SCC values of cows in the probiotic group were significantly lower than those in the control group. In the early lactation period (days in milk 0-150), the body condition score of cows improved after supplementing with probiotics. Additionally, milk yield and lactation persistency increased post-partum, with significant increases in milk protein and casein content. Regarding the possible mechanisms, we explored rumen fermentation and microbiota. In terms of total volatile fatty acids (VFA) content, there was a significant increase in the T1 stage at the Taipei farm, while a significant decrease was observed in the S4 stage at the Tainan farm. The changes in the proportions of various VFAs were not consistent, but both farms observed a increase in isobutyric acid and microbial crude protein (MCP) content following probiotic supplementation. This increase may indicate that probiotics promote effective metabolism and utilization of amino acids in the rumen. In terms of microbiota, probiotic supplementation significantly increased microbial diversity in the rumen. We observed that probiotics increased the relative abundance of Ruminococcus and Bifidobacterium, which are biomarkers for healthy lactating cows. Additionally, we found that probiotic supplementation increased the content of Breznakia pachnodae and Sodaliphilus pleomorphus, which are positively correlated with MCP synthesis. Functional prediction analysis showed a significant increase in amino acid and carbohydrate metabolism pathways in the rumen microbiota of cows supplemented with probiotics, suggesting that probiotics can improve digestion and absorption of nutrients by influencing the rumen microbiota and its metabolic activities, thereby enhancing health and production performance. For calves, the incidence of diarrhea significantly decreased after probiotic supplementation. In terms of microbiota, we found an increase in Pseudoramibacter, Pseudoramibacter porci, and Solibacillus isronensis B3W22, which are negatively correlated with fecal scores. Gene function prediction observed an increase in amino acid metabolism and signal transduction pathways, indicating that probiotics may improve digestion, absorption, and immune function by modulating the gut microbiota, thereby enhancing disease resistance in calves. In summary, this study confirms that the novel probiotic combination has a positive impact on the health status and production performance of dairy cows and calves in different farm environments, and it has practical application potential in the livestock industry for reducing antibiotic use, increasing economic benefits and enhancing the welfare of dairy cows.