Figure 2 - available via license: Creative Commons Attribution 3.0 Unported
Content may be subject to copyright.
Amperometric immunosensor for lactoferrin determination using hydrogen peroxide electrode as transducer.
Source publication
Two different immunosensors, recently developed for the determination of antibacterial proteins (lactoferrin and immunoglobulin G) in buffalo milk and in other commercial animal milks samples, were used in the present study. The aim was to propose these immunosensor methods for routine control of important diet products, such as cow and goat milks,...
Contexts in source publication
Context 1
... Immobilon Ny+ Membrane was cut into approximately 1 cm surface area disks and 100 μL of a 1.0 mg/mL anti-lactoferrin was directly deposited on the membrane surface. The membrane was then dried at room temperature for about 24 h and stored at 4° C. The transducer consisted of an amperometric electrode for H 2 O 2 determination, with a Pt anode and an Ag/AgCl/Cl - cathode, provided with a plastic cap filled with 0.1 M KCl solution and screwed onto the body of the electrode, at the lower end of which a dialysis membrane was positioned. The Immobilon membrane with the immobilized anti-lactoferrin overlapped the dialysis membrane. Finally, a nylon net overlapped the latter membrane. The two membranes and the net were secured by a rubber O-ring to the plastic cap of the electrode as shown in Figure ...
Similar publications
The objective of this study was to quantify lactoferrin (Lfe) in buffalo milk and to examine the factors affecting milk Lfe, such as the lactation stage, daily milk yield, parity, and milk somatic cells count (SCC). Milk Lfe concentration was detected by the SDS-polyacril-amide gel electrophoresis (SDS-PAGE). The overall mean of Lfe concentration w...
Citations
... Colostrum represents a critical source of immune-protection for newborn calves [2]. As ruminants, buffalo calves are born agammaglobulemic and highly dependent on efficient enteral uptake of maternal immunoglobulin G (IgG) from colostrum to protect them from infections until their own immune system has developed [3][4][5]. ...
Brix refractometry has been widely demonstrated to be a useful tool for monitoring colostrum management program and passive immunity transfer (PIT) in Bovines, but its suitability has never been verified in Buffalo. Therefore, the objective of this study was to evaluate the utility of a simple and rapid tool such as a digital Brix refractometer to estimate colostrum quality and for evaluating the success of passive transfer of immunoglobulin G (IgG) in Buffalo calves. The optimal cut points levels for Brix Refractometry for distinguishing good- and poor-quality colostrum and for assessing the adequacy of passive immunity transfer in calves were determined. For this aim, 26 first-milking maternal colostrum (MC) were collected from first-calf heifers. Blood samples were obtained from their calves at birth (T0) and 72 hours after (T3). Colostrum and Serum IgG content were determined by indirect enzyme-linked immunosorbent assay (ELISA), whereas total protein (TP, g/dL) and percentage Brix (%Brix) by means of a digital Brix refractometer. The mean colostrum IgG was 64.9 ± 29.3 mg/mL. The mean serum %Brix at T3 was 9.6 ± 0.9 %. The mean serum IgG content at T3 was 11.1 ± 2.0 mg/mL. Pearson’s correlation coefficient (rp) was determined between Brix and ELISA measurements: colostrum %Brix showed a significant correlation with serum %Brix (rp = 0.82, p < 0.001); serum %Brix was highly correlated with serum TP (STP, g/dL) (rp = 0.98, p < 0.001) and serum IgG (mg/mL) (rp = 0.85, p < 0.001). A cut point of 18% Brix to estimate samples of MC ≥ 50 mg/mL from first-calf heifers was more appropriate for the buffalo. A cut point of 8.4% Brix resulted in the greatest percentage of calf serum samples being correctly classified. Based on our findings, a digital Brix refractometer could be a useful tool to monitor colostrum quality and to estimate PIT in Buffalo calves.
... A proteína IgG é conhecida por fazer parte da imunidade passiva, como proteína de defesa, transferida do leite colostral ao bezerro. Auxilia, juntamente com a proteína LF, na proteção do neonato contra infecções, até que consiga desenvolver seu próprio sistema imunológico (Campanella et al., 2009;Hurley & Theil, 2011). A elevação da concentração da IgG em amostras de leite de vacas com mastite vem sendo observada em outros estudos (Harmon et al., 1976;Ostensson & Lun, 2008). ...
Objetivou-se, com o presente estudo, investigar a influência do aumento da CCS na expressão das proteínas do soro de leite por meio da utilização da técnica de separação de proteínas de eletroforese microfluídica em microchip (lab-on-a-chip). Foram coletadas 104 amostras de leite de vacas em duas propriedades localizadas no Estado de Goiás. As amostras foram analisadas quanto a contagem de células somáticas e eletroforese microfluídica lab-on-a-chip. Para o teste de correlação entre o perfil de proteínas e a CCS foram utilizadas 50 amostras de leite e estas foram estratificadas em cinco grupos de escores de células somáticas, sendo utilizado um delineamento inteiramente ao acaso com cinco tratamentos, avaliado por meio da estatística descritiva e representações gráficas. Verificou-se um aumento significativo no valor do componente proteína em relação aos ECS 1 e 3, 1 e 4 e 1 e 5. Também foi possível observar diferença estatística nos resultados do componente Lactose quando se comparou os ECS 1 e 3 e 3 e 4. Na avaliação descritiva da variável “Concentração” das proteínas do leite foi possível observar diferenças nos valores das concentrações em função do ECS. Foi possível observar diferença estatística para os resultados obtidos para as proteínas α-LA, quando comparados os ECS 1, 2 e 3 e o ECS 5; para a IgG, as amostras de ECS 4 e 5 apresentaram concentrações diferentes das amostras dos ECS 1, 2 e 3; e a proteína LF, apresentou resultados diferentes entre as amostras de ECS 1 e 5. Concluiu-se que proteínas de alta abundância do soro de leite tem sua concentração diminuída em função da severidade da mastite; proteínas de defesa, como Lactoferrina, Lactoperoxidase e IgG e IgM têm aumento em suas concentrações quando se comparam amostras de leite de vacas com mastite subclínica e amostras de leite de vacas sadias; as proteínas Lactoferrina e IgG constituem alvos em potencial para identificar vacas com mastite subclínica a partir de amostras de leite, podendo ser consideradas biomarcadores.
... Cow's milk is the most readily available source of lactoferrin, with an average concentration of 0.174 g/l in low heat pasteurized cow's milk (and 1.2 g/kg in semihard cheese produced from that milk) [129], which is in good agreement with other studies [130][131][132], though the range experienced (0.03-0.486 g/l) is dependent on several factors [133]. The concentration in colostrum is higher, but varies greatly between breeds and may be anything between 0.3 and 5 g/l, and is typically at the lower end of the range [134][135][136]. ...
Coronavirus disease 2019 (COVID-19), caused by SARS coronavirus 2 (SARS-CoV-2), is spreading rapidly with no established effective treatments. While most cases are mild, others experience uncontrolled inflammatory responses with oxidative stress, dysregulation of iron and coagulation as features. Lactoferrin, ovotransferrin and lysozyme are abundant, safe antimicrobials that have wide antiviral as well as immunomodulatory properties. In particular, lactoferrin restores iron homeostasis and inhibits replication of SARS-CoV, which is closely related to SARS-CoV-2. Ovotransferrin has antiviral peptides and activities that are shared with lactoferrin. Both lactoferrin and lysozyme are ‘immune sensing’ as they may stimulate immune responses or resolve inflammation. Mechanisms by which these antimicrobials may treat or prevent COVID-19, as well as sources and forms of these, are reviewed.
... Systematic breeding studies related to Anatolian buffaloes have just been started in Turkey. Although many studies report the lactoferrin concentration in milk obtained from cows before the year 2000 (Shimazaki et al., 2017), there is limited information on the determination of lactoferrin levels in buffalo milk (Elagamy, 2000;Campanella et al., 2009;Giacinti et al., 2013). However, there is no comparative analysis of lactoferrin levels between bacterial agents and milk lactoferrin levels in buffaloes with subclinical mastitis. ...
... In milk samples obtained from five healthy buffaloes in Italy, milk lactoferrin levels ranged from 5.6 µg/ml to 800 µg/ml while the mean concentration was 232 µg/ml (Campanella et al., 2009). In another report, Giacinti et al. (2013) showed that the milk lactoferrin level ranged from 30 mg/kg to 813 mg/kg in buffaloes, with a mean value of 332 mg/kg. ...
... The mean lactoferrin levels in tank milk from cows and buffaloes were indicated as being, respectively, 76.7 µg/ml and 31.7 µg/ ml (Elagamy, 2000). In this study, the mean milk lactoferrin level in the healthy mammary quarters (27.86±1.65 µg/ml) was similar to that of Elagamy (2000), but lower than in the other reports (Campanella et al., 2009;Giacinti et al., 2013). ...
This study aimed to determine the lactoferrin concentration from healthy and subclinical mastitic buffaloes. Lactoferrin levels were determined in a total of 475 mammary quarter milk samples, 391 of which were healthy and 84 suffering from subclinical mastitis. The mean milk lactoferrin level in logarithmic form for healthy and subclinical mastitic milk samples was 1.34±0.27 and 1.85±0.47, respectively. A significant difference (P<0.001) was determined between the two groups in terms of the lactoferrin levels. The milk lactoferrin levels in healthy buffaloes were significantly higher in the middle and late periods of lactation than the peak period (P<0.05). The mean milk lactoferrin levels in subclinical mastitic buffaloes infected with S. aureus or S. agalactiae were significantly higher than those from in buffaloes infected with Coagulase-Negative Staphylococci (CNS) or Candida spp. (P<0.05). An increase in milk lactoferrin levels was observed in parallel with the increase in somatic cell scores. In conclusion, milk lactoferrin levels in buffaloes were found to increase with the lactation stage, the presence of bacterial infection and the somatic cell count. This is the first study to determine the relationship between lactoferrin concentrations and bacterial infection in milk obtained from mammary quarters with subclinical mastitis in Anatolian buffaloes.
... Bovine, goat and camel milks were the most investigated ones [14,15]. Milk includes numerous compounds such as lacto-peroxidase, lactoferrin, immunoglobulin G, secretory immunoglobulin A, and Lysozymes [16]. The protective effect of these proteins had been screened against several bacterial strains like Escherichia coli, Staphylococcus aureus, Salmonella typhimurium, Lactococcus lactis and rotavirus [17]. ...
... Antihypertensive, antimicrobial, antiinflammatory and Inmunostimulating activities Important source of amino acids (Lacomba et al., 2010;Campanella et al., 2009;Meltretter et al., 2008;De Simone et al., 2009) Amino acids Sprouts alga and sport drinks and tablets ...
Healthy nutrition is important for human beings for good health provided by global industry. Nutraceutical and functional food provide a prospect to reduce health care costs and improve the human health. Researchers carried out plenty of work for the preparation of nutraceutical and functional food product. This article also focused on recent advances on the nutraceutical and functional foods product on health benefits and their application in prevention of disease. Here we discussed about the health benefit of recently introduced nutraceutical and functional food products. With the modernised, competitive lifestyle and ever increasing stressful conditions this product is the need of the day.
... In contrast, Sant'ana et al [17] did not observe any significant variation in Lfe content during lactation. Campanella et al and Giacinti et al [18,19] in analyzing Lfe concentrations in raw buffalo milk found values of 232 μg/mL and 332 μg/mL, respectively. This difference in the average content of Lfe among the studies suggests the influence of the animal' s age, the lactation period and the methodology used. ...
Objective:
This study aimed to evaluate the microbiological and cellular milk profile for the diagnosis of subclinical mastitis in female buffaloes and to assess risk factors for predisposition of the disease.
Methods:
Analyses were carried out by standard plate count (SPC), identification of species and antibiotic resistance, somatic cell count (SCC), electrical conductivity of milk (ECM), and Lactoferrin content in milk (Lfe). Teat cups were swabbed to evaluate risk factors, observing hyperkeratosis, milking vacuum pressure and cleanliness of the site. Hence, 30 female buffaloes were randomly selected (15 from a group in early lactation and 15 in late lactation).
Results:
The most common bacteria in the microbiological examination were Staphylococcus spp., Streptococcus spp. and Corynebacterium sp. In the antibiotic sensitivity test, 10 (58.82%) of the 17 antibiotics tested were sensitive to all isolates, and resistant bacteria were S. uberis, S. dysgalactiae, S. haemolyticus and E. coli. It was observed that positive samples in the microbiological examination showed total bacterial count (TBC) between 9.10 x 103 to 6.94 x 106 CFU/mL, SCC between 42,000 to 4,320,000 cells/mL and ECM ranging from 1.85 to 7.40 mS/cm. It was also found that the teat cups had high microbial counts indicating poor hygiene, and even faults in the cleanliness of the animals' waiting room were observed. It is concluded that values of SCC above 537,000 cells/mL and ECM above 3.0 mS/mL are indications of mammary gland infection for this herd; however, the association of these values with a microbiological analysis is necessary to more accurately evaluate the health status of mammary glands with subclinical mastitis.
Conclusion:
Through phenotypic characterization of bacteria involved in the samples, the genera Staphylococcus spp., Streptococcus spp. and Corynebacterimum bovis were the most prevalent in this study. Faults in environment and equipment hygienization are factors that are directly associated with mastitis.
... Both the biosensor methods have been previously described in detail [10][11][12][13]. ...
... For the lactoferrin analysis of liquid samples, all milk samples and the dietary supplement n. 1 were analyzed without dilution, i.e. 5 mL of milk (or 1 vial of the integrator n. 1, containing 8 mL of product), were added directly into the measuring cell; while for the pharmaceutical integrators n. 2 and n. 3, the content of a sachet was weighed and dissolved in 5 mL of 0.1 M phosphate buffer pH 7.5 and analyzed with the format reported in detail in previous papers [10,13]. For the analysis of recoveries in expired milk, or in three different integrators, samples were also analyzed as they were, after dissolution and homogenization, such above described, or after proper dilution. ...
... Using two sensors, optimized in previous researches [10][11][12][13], we simultaneously measured lactoferrin concentration and antioxidant capacity in three food supplements sold in drugstores (see Table 1) as well as in cow, goat, powdered milk and in yoghurt. The validation of two biosensor methods has been reported and discussed in previous published papers [10][11][12][13]. ...
Background: Recently also food supplements for adults containing lactoferrin have appeared on the market. Recent experimental results support the proposal that oral supplements of bovine lactoferrin may be a useful adjunct in the modulation of immune activity, in particular T-cell activation and antioxidant status. Objective: The present research shows as it is possible to determine in a successfully and inexpensive way, respectively using an immunosensor and an enzyme sensor, both the lactoferrin content, and the total antioxidant capacity values in real samples. Food samples analyzed to this purpose have been food integrators containing lactoferrin, available in drug stores, edible animal milk samples and dairy derivatives, such as powdered milk, or yogurt. Method: Using two sensors, optimized in previous researches, we simultaneously measured lactoferrin concentration and antioxidant capacity in food supplements sold in drugstores as well as in cow, goat, powdered milk and in yoghurt. Results: It was also tried to verify if exists between found lactoferrin content and total antioxidant capacity data values, any correlation. Actually some correlation has been found in milk samples, but only in one of the analyzed food integrators this correlation has been evidenced. Results emphasize as the lactoferrin content was clearly shown to contribute to the total antioxidant capacity in both food supplements and various milk and milk derivative samples. Conclusion: The present research also proved that it was possible to perform a suitable and fast determination of lactoferrin in the food supplements tested and to compare the lactoferrin concentration, declared by the manufacturers, with the experimental measure content value, using an inexpensive immunosensor, instead other more complex and costly instrumental methods. At this stage of the research it was not always possible to prove a direct correlation between lactoferrin concentration and (TAOC) value in all examined food supplements, as some other molecules with a well known high antioxidant capacity are present in the analyzed food supplements.
... More recently, enzymatic and nephelometric immunoassays for lactoferrin in milk have been reported (221)(222)(223)(224)(225)(226). Conventional immunoassay techniques have been further enhanced through the development of immunosensor platforms exploiting electrochemical transduction techniques, which have been applied to the determination of lactoferrin in mammalian milk (227,228). Alternative real-time, label-free optical biosensor techniques based on SPR transduction have also been reported for the quantitation of lactoferrin in bovine milk and infant formula milk (194,229). Such automated biosensor immunoaffinity technologies exploit the specific and reversible interaction between antibody and protein antigen, and they are versatile, robust, and capable of producing rapid and reliable data for the quantitative analysis of lactoferrin, and other proteins, in complex food matrices with minimal sample preparation. ...
Infant formula is designed to provide the human infant with a sole source of nutrition and it is intended to imitate breast milk. In recent years, advances in the science of infant nutrition have led to an increasing number of novel ingredients that are supplemented into infant formula. As the list of these nutritionally important nutrients is lengthy, this review summarizes contemporary analytical methods that have been applied to a representative selection (lutein, carnitine, choline, nucleotides, inositol, taurine, sialic acid, gangliosides, triacylglycerides, oligosaccharides, α-lactalbumin, and lactoferrin).
... actation it was observed that lactoferrin was more than the following lactation days. Thus it can be seen that lactation period, age and other maternal characteristics plays important role in the lactoferrin The alteration of activity of lactoferrin in milk could have an impact on the shelf life of raw milk and also on the development of neonates (Campanella et. al., 2009). Presence of glycan molecule in the structure of lactoferrin prevents degradation of itself by proteolytic enzymes such as trypsin and trypsin like enzymes which facilitates partial resistance to digestion in the gut. Lactoferrin is considered to be an important host defence molecule and has a diverse range of physiological activities ...
Milk is a highest quality source of well balanced nutrients and also displays a range of biological activities that affects digestion, metabolic responses to absorbed nutrients, growth & development of specific organs, and resistance to disease. Bioactive proteins such as lactoferrin (Lf) have been isolated over decades ago and showed their importance in stimulating immune system in the infants through breast milk in addition to immunoglobulin present in the milk. In addition to immune system stimulation, Lf also has antibacterial activity and antioxidant activity in infant and adult of human as well as animal health. In this review paper, antibacterial properties of lactoferrin have been discussed along with its future perspectives.
