Evaluation of a povidone-iodine and chitosan-based barrier teat dip in the prevention of mastitis in dairy cows

doi:10.1016/S2095-3119(20)63418-9

摘要/Abstract

摘要：

后药浴剂是预防奶牛乳腺炎的有效手段之一，本研究旨在探究一种基于聚维酮碘-壳聚糖的乳头成膜药浴剂对奶牛乳腺炎的预防效果，主要包括以下几个方面：一、壳聚糖对奶牛乳腺炎致病菌抑菌效果的研究：利用最低抑菌浓度实验考察不同分子量壳聚糖（5，50，150，350 kDa）对六种常见乳腺炎致病菌的抑菌效果，结果显示50 kDa的壳聚糖对六种致病菌的抑菌效果最好，将其应用于下一步的药浴剂制备；二、壳聚糖添加量对成膜药浴剂抑菌性能的影响：利用药敏实验考察不同壳聚糖添加量（0.0, 0.5, 1.0, 1.5%）对六种常见乳腺炎致病菌的抑菌效果，结果显示含4%聚维酮碘及1%壳聚糖的成膜药浴剂对大肠杆菌、金黄色葡萄球菌、无乳链球菌、蜡样芽胞杆菌的抑菌效果要显著大于含4%聚维酮碘的成膜药浴剂（P<0.05）；三、成膜药浴剂临床效果研究-乳区实验：选取47头健康中国荷斯坦牛进行为期56天的乳区对比实验，奶牛左侧2个乳区后药浴使用含4%聚维酮碘及1%壳聚糖的成膜药浴剂，右侧2个乳区后药浴使用10%聚维酮碘。在试验开始时（0天）和第28、56天时采集奶样，进行乳成分和乳中体细胞数（Somatic Cell Count，SCC）测定，同时针对SCC高于200,000个/mL的奶样进行致病菌的分离鉴定。结果表明：左侧乳区和右侧乳区奶样在0、28、56天时SCC及乳成分等指标均无显著差异（P>0.05）。四、成膜药浴剂临床效果研究-牛群实验：选取139头健康中国荷斯坦奶牛进行为期56天的群体对比实验，其中对照组67头，实验组72头。对照组后药浴使用10%聚维酮碘，实验组后药浴使用聚维酮碘-壳聚糖成膜药浴剂。结果表明：56天时，实验组的隐性乳腺炎发病率比对照组降低了29% (P<0.05)。在实验组内，第56天的隐性乳腺炎发病率低于0天 (P<0.05)。此外，实验组和对照组奶样在0、28、56天时SCC、乳成分指标和临床乳腺炎发病率均无显著差异（P>0.05）。微生物分离鉴定实验结果表明基于聚维酮碘-壳聚糖的乳头成膜药浴剂可有效预防由肺炎克雷伯菌、铜绿假单胞菌和弗格森大肠杆菌引起的隐性乳腺炎。综上所述，含4%聚维酮碘及1%壳聚糖的成膜药浴剂预防奶牛隐性乳腺炎的效果要优于10%聚维酮碘，该研究为扩大壳聚糖在奶牛乳腺炎防治方面的应用，开发壳聚糖相关的绿色兽药产品提供了科学的理论依据。

Abstract:

Postmilking teat dip is an important tool used to prevent mastitis in the modern dairy industry. In this study, we evaluated the in vitro and in vivo efficacies of a barrier teat dip containing povidone-iodine and chitosan for the prevention of mastitis. In experiment 1, we evaluated the antibacterial effects of chitosans with different molecular weights against six mastitis-causing bacteria based on the minimal inhibitory concentration test. The results showed that 50 kDa chitosan had the maximum antibacterial activity compared with 5, 150 and 350 kDa chitosans. In experiment 2, the inhibition zone test indicated that the barrier teat dip with 4.0% povidone-iodine and 1.0% chitosan had higher (P<0.05) in vitro antibacterial efficacy against most tested mastitis-causing bacteria than the barrier teat dip with 4.0% povidone-iodine and no chitosan. In experiments 3 and 4, we evaluated the efficacies of two postmilking teat dips, 1) a barrier teat dip containing 1.0% chitosan and 4.0% povidone-iodine and 2) a conventional nonbarrier product containing 10% povidone-iodine in a field trial at two commercial dairy herds (1 and 2). A 56-d split-udder experiment (experiment 3) was conducted using 47 lactating Chinese Holstein cows in herd 1. Both left teats were immersed in barrier postmilking dip, and both right teats were dipped with nonbarrier postmilking dip. During a 56-d split-herd experiment (experiment 4), a total of 139 lactating Chinese Holstein cows from herd 2 were allocated to two groups: 1) all teats of 67 cows were dipped in the nonbarrier teat dip, and 2) all teats of 72 cows were dipped in the barrier teat dip. Milk samples were collected and analyzed for somatic cell count (SCC), fat content, protein content, and fat-to-protein ratio prior to the start of sampling (0 d), and at 28 and 56 d after initiation. Bacteriological analysis was only performed on milk samples with SCC≥200?000 cells mL^–1. In experiment 3, no differences (P>0.05) in SCC, somatic cell score (SCS) or other milk quality indicators were observed between nonbarrier and barrier teat dip treatment teats throughout the experiment. At the end of experiment 4, compared with nonbarrier teat dip group, a reduction (P<0.05) of 29% was observed for subclinical mastitis infection prevalence in the barrier teat dip group. In the barrier teat dip group, the subclinical mastitis infection prevalence on 56 d was lower (P<0.05) than 0 d. No differences (P>0.05) in milk qualities or clinical mastitis incidence were detected between groups. Bacteriological analysis demonstrated that the barrier product containing povidone-iodine and chitosan reduced the subclinical mastitis infection prevalence induced by mastitis pathogens. This effect was mainly due to the reductions in Klebsiella pneumoniae, Pseudomonas aeruginosa, and Escherichia fergusonii infections. Overall, the data indicated that a barrier teat dip containing 4% povidone-iodine and 1% chitosan was more effective than 10% povidone-iodine in preventing subclinical mastitis.

Key words: mastitis , povidone-iodine , chitosan , barrier postmilking dip

. [J]. Journal of Integrative Agriculture, 2021, 20(6): 1615-1625.

ZHANG Hui-min, JIANG Hong-rui, CHEN Dai-jie, SHEN Zi-liang, MAO Yong-jiang, LIANG Yu-sheng, Juan J. LOOR, YANG Zhang-ping. Evaluation of a povidone-iodine and chitosan-based barrier teat dip in the prevention of mastitis in dairy cows[J]. Journal of Integrative Agriculture, 2021, 20(6): 1615-1625.

参考文献

Asli A, Brouillette E, Ster C, Ghinet M G, Brzezinski R, Lacasse P, Jacques M, Malouin F. 2017. Antibiofilm and antibacterial effects of specific chitosan molecules on Staphylococcus aureus isolates associated with bovine mastitis. PLoS ONE,12, e0176988.
Bain M S, Green C C. 1999. Isolation of Escherichia fergusonii in cases clinically suggestive of salmonellosis. The Veterinary Record, 144, 511–511.
Banerjee S, Batabyal K, Joardar S N, Isore D P, Dey S, Samanta I, Samanta T K, Murmu S. 2017. Detection and characterization of pathogenic Pseudomonas aeruginosa from bovine subclinical mastitis in West Bengal, India. Veterinary World, 10, 738–742.
El Behiry A, Schlenker G, Szabo I, Roesler U. 2012. In vitro susceptibility of Staphylococcus aureus strains isolated from cows with subclinical mastitis to different antimicrobial agents. Journal of Veterinary Science, 13, 153–161.
Breser M L, Felipe V, Bohl L P, Orellano M S, Isaac P, Conesa A, Rivero V E, Correa S G, Bianco I D, Porporatto C. 2018. Chitosan and cloxacillin combination improve antibiotic efficacy against different lifestyle of coagulase-negative Staphylococcus isolates from chronic bovine mastitis. Scientific Reports, 8, 5081.
Breen J E, Bradley A J, Green M J. 2009. Quarter and cow risk factors associated with a somatic cell count greater than 199 000 cells per milliliter in United Kingdom dairy cows. Journal of Dairy Science, 92, 3106–3115.
Chen L C, Kung S K, Chen H H, Lin S B. 2010. Evaluation of zeta potential difference as an indicator for antibacterial strength of low molecular weight chitosan. Carbohydrate Polymers, 82, 913–919.
Chen Y, Yang Y M, Liao Q P, Yang W, Ma W F, Zhao J, Zheng X G, Yang Y, Chen R. 2016. Preparation, property of the complex of carboxymethyl chitosan grafted copolymer with iodine and application of it in cervical antibacterial biomembrane. Materials Science & Engineering (C: Materials for Biological Applications), 67, 247–258.
Davoodbasha M, Lee S Y, Kim J W. 2018. Solution plasma mediated formation of low molecular weight chitosan and its application as a biomaterial. International Journal of Biological Macromolecules, 118, 1511–1517.
Dore S, Ferrini A M, Appicciafuoco B, Massaro M R, Sotgiu G, Liciardi M, Cannas E A. 2019. Efficacy of a terpinen-4-ol based dipping for post-milking teat disinfection in the prevention of mastitis in dairy sheep. Journal of Essential Oil Research, 31, 19–26.
Foret C, Aguro A, Janowicz P. 2006. Efficacy of two barrier iodine teat dips under natural exposure conditions. Journal of Dairy Science, 89, 2279–2285.
Fuenzalida M J, Ruegg P L. 2020. Molecular epidemiology of nonsevere clinical mastitis caused by Klebsiella pneumoniae occurring in cows on 2 Wisconsin dairy farms. Journal of Dairy Science, 103, 3479–3492.
Hillerton J E, Cooper J, Morelli J. 2007. Preventing bovine mastitis by a postmilking teat disinfectant containing acidified sodium chlorite. Journal of Dairy Science, 90, 1201–1208.
Kelly E J, Wilson D J. 2016. Pseudomonas aeruginosa mastitis in two goats associated with an essential oil-based teat dip. Journal of Veterinary Diagnostic Investigation, 28, 760–762.
Kong M, Chen X G, Xing K, Park H J. 2010. Antimicrobial properties of chitosan and mode of action: A state of the art review. International Journal of Food Microbiology, 144, 51–63.
Lanctot S, Fustier P, Taherian A R, Bisakowski B, Zhao X, Lacasse P. 2017. Effect of intramammary infusion of chitosan hydrogels at drying-off on bovine mammary gland involution. Journal of Dairy Science, 100, 2269–2281.
Martins C, Pinheiro E S C, Gentilini M, Benavides M L, Santos M V. 2017. Efficacy of a high free iodine barrier teat disinfectant for the prevention of naturally occurring new intramammary infections and clinical mastitis in dairy cows. Journal of Dairy Science, 100, 3930–3939.
Munoz M A, Welcome F L, Schukken Y H, Zadoks R N. 2007. Molecular epidemiology of two Klebsiella pneumoniae mastitis outbreaks on a dairy farm in New York State. Journal of Clinical Microbiology, 45, 3964–3971.
NRC (National Research Council). 2001. Nutrient Requirements of Dairy Cattle. 7th ed. National Academies Press, Washington, D.C.
Peng Y B, Song C L, Yang C F, Guo Q G, Yao M. 2017. Low molecular weight chitosan-coated silver nanoparticles are effective for the treatment of MRSA-infected wounds. International Journal of Nanomedicine, 12, 295–304.
Raafat D, von Bargen K, Haas A, Sahl H G. 2008. Insights into the mode of action of chitosan as an antibacterial compound. Applied and Environmental Microbiology, 74, 3764–3773.
Raafat D, Sahl H G. 2009. Chitosan and its antimicrobial potential - A critical literature survey. Microbial Biotechnology, 2, 186–201.
Rahaman M S, Rahman M M, Moghal M M R, Rakib T M, Siddiki A M A M Z. 2017. Characterization of bacteria causing bovine mastitis by amplified ribosomal DNA restriction analysis (ARDRA) method. Bangladesh Journal of Veterinary and Animal Sciences, 5, 44–51.
Ruegg P L. 2017. A 100-Year Review: Mastitis detection management and prevention. Journal of Dairy Science, 100, 10381–10397.
Saber A, Strand S P, Ulfendahl M. 2010. Use of the biodegradable polymer chitosan as a vehicle for applying drugs to the inner ear. European Journal of Pharmaceutical Sciences, 39, 110–115.
Schepers A J, Lam T, Schukken Y H, Wilmink J B M, Hanekamp W J A. 1997. Estimation of variance components for somatic cell counts to determine thresholds for uninfected quarters. Journal of Dairy Science, 80, 1833–1840.
Silva-Dias A, Palmeira-De-Oliveira A, Miranda I M, Branco J, Cobrado L, Monteiro-Soares M, Queiroz J A, Pina-Vaz C, Rodrigues A G. 2014. Anti-biofilm activity of low-molecular weight chitosan hydrogel against Candida species. Medical Microbiology and Immunology, 203, 25–33.
Tokura S, Ueno K, Miyazaki S, Nishi N. 1996. Molecular weight dependent antimicrobial activity by chitosan. In: New Macromolecular Architecture and Functions. Springer, Berlin Heidelberg. pp. 199–207.
Zemljic L F, Persin Z, Sauperl O, Rudolf A, Kostic M. 2018. Medical textiles based on viscose rayon fabrics coated with chitosan-encapsulated iodine: Antibacterial and antioxidant properties. Textile Research Journal, 88, 2519–2531.
Zheng L Y, Zhu J A F. 2003. Study on antimicrobial activity of chitosan with different molecular weights. Carbohydrate Polymers, 54, 527–530.
Zohri M, Alavidjeh M S, Haririan I, Ardestani M S, Ebrahimi S E S, Sani H T, Sadjadi S K. 2010. A comparative study between the antibacterial effect of nisin and nisin-loaded chitosan/alginate nanoparticles on the growth of Staphylococcus aureus in raw and pasteurized milk samples. Probiotics and Antimicrobial Proteins, 2, 258–266.