Aims &Objective: Mycobacterium bovis is the causative agent of bovine tuberculosis. It is also a pathogen for other animals such as goats and humans. Molecular methods are applied to detect members ...of the M. tuberculosis complex in different clinical samples as milk. The Loop-Mediated Isothermal Amplification (LAMP) proceeds at a constant temperature using a strand displacement reaction. LAMP does not require a thermal cycler. It is characterized by its efficiency, rapidity, high yield of final product, robustness, sensitivity and specificity. The primers are specifically designed to recognize 6 distinct regions on the target gene. The amplified product can be detected by naked eye as a white precipitate or a color change of the reaction solution.: To detect species of the Mycobacterium tuberculosis complex in goat milk samples by IS6110-LAMP
Methods: Twenty goat milk samples from animals reacting to the tuberculin test were studied by IS6110-LAMP, Touch-Down (TD) IS6110 and Rv2807 PCR. These goats had been slaughtered, their organs and milk cultured. The isolates obtained were typed by spoligotyping. DNA extraction was performed using 500uL of milk sample. It was incubated with sodium dodecyl sulfate / proteinase K and treated twice with phenol:chloroform:isoamylic alcohol and once with chloroform:isoamylic alcohol. Then NaCl was added and DNA was precipitated with isopropanol and washed with ethanol. The remaining ethanol was evaporated and the pellet resuspended in water. IS6110-LAMP reaction was performed in a 25uL volume containing Bst DNA polymerase, betaine, magnesium sulfate, dNTPs, and primers (F3, B3, FIP, BIP, LF, LR). SYBR Gold solution was added inside of the cup of the tubes that were incubated at 63°C during one hour. After that the tubes were shacked and a yellowish color indicated a positive reaction.
The specificity of IS6110-LAMP was evaluated using DNA from different species of Non Tuberculous Mycobacterias (NTM) (Mycobacterium smegmatis, M. phlei, M. fortuitum, M. gordonae, M. intracellulare, M. porcinum, M. avium subsp. avium, M. avium subsp. paratuberculosis) and the close related Nocardia sp. TD IS6110 and TDRv2807 PCR were also performed with an initial step of 96°C for 3 min, followed by 8 cycles of 96°C for 1 min, annealing temperatures starting at 72°C for 1 min (decreasing by 1 °C/cycle), and 72°C for 1 min for extension. This step was followed by 30 cycles of 96°C for 1 min, 65°C for 1 min, 72°C for 2 min, and a final extension at 72°C for 8 min. The amplification products were 245 and 443 bp, respectively.
Results: While 17/20 goats were confirmed by bacteriology, no isolates were obtained from milk culture. The 17 isolates were further confirmed as M. bovis by spoligotyping. Neither NTM nor Nocordia sp. templates amplified by IS6110-LAMP. The 40 % (8/20) of the samples were IS6110-LAMP positive. There was only one discordant result between IS6110-LAMP, TD-Rv2807 and TD-IS6110.
Conclusion: IS6110-LAMP is a useful tool to detect species of the M. tuberculosis complex in goat milk samples that could be implemented in laboratories with low technological complexity.
Consumer perception of organic cow milk is associated with the assumption that organic milk differs from conventionally produced milk. The value associated with this difference justifies the premium ...retail price for organic milk. It includes the perceptions that organic dairy farming is kinder to the environment, animals, and people; that organic milk products are produced without the use of antibiotics, added hormones, synthetic chemicals, and genetic modification; and that they may have potential benefits for human health. Controlled studies investigating whether differences exist between organic and conventionally produced milk have so far been largely equivocal due principally to the complexity of the research question and the number of factors that can influence milk composition. A main complication is that farming practices and their effects differ depending on country, region, year, and season between and within organic and conventional systems. Factors influencing milk composition (e.g., diet, breed, and stage of lactation) have been studied individually, whereas interactions between multiple factors have been largely ignored. Studies that fail to consider that factors other than the farming system (organic vs. conventional) could have caused or contributed to the reported differences in milk composition make it impossible to determine whether a system-related difference exists between organic and conventional milk. Milk fatty acid composition has been a central research area when comparing organic and conventional milk largely because the milk fatty acid profile responds rapidly and is very sensitive to changes in diet. Consequently, the effect of farming practices (high input vs. low input) rather than farming system (organic vs. conventional) determines milk fatty acid profile, and similar results are seen between low-input organic and low-input conventional milks. This confounds our ability to develop an analytical method to distinguish organic from conventionally produced milk and provide product verification. Lack of research on interactions between several influential factors and differences in trial complexity and consistency between studies (e.g., sampling period, sample size, reporting of experimental conditions) complicate data interpretation and prevent us from making unequivocal conclusions. The first part of this review provides a detailed summary of individual factors known to influence milk composition. The second part presents an overview of studies that have compared organic and conventional milk and discusses their findings within the framework of the various factors presented in part one.
Objectives To prospectively evaluate the effect of different dietary management strategies on the rate of acquisition of tolerance in children with cow's milk allergy (CMA). Study design Otherwise ...healthy children (aged 1-12 months) diagnosed with CMA were prospectively evaluated. The study population was divided into 5 groups based upon the formula used for management: (1) extensively hydrolyzed casein formula (EHCF, n = 55); (2) EHCF + Lactobacillus rhamnosus GG LGG, n = 71); (3) hydrolyzed rice formula (RHF, n = 46); (4) soy formula (n = 55); and (5) amino acid based formula (n = 33). A food challenge was performed after 12 months to assess acquisition of tolerance. Results Two hundred sixty children were evaluated (167 male, 64.2%; age 5.92 months, 95% CI 5.48-6.37; body weight 6.66 kg, 95% CI 6.41-6.91; IgE-mediated CMA 111, 42.7%). The rate of children acquiring oral tolerance after 12 months was significantly higher ( P < .05) in the groups receiving EHCF (43.6%) or EHCF + LGG (78.9%) compared with the other groups: RHF (32.6%), soy formula (23.6%), and amino acid based formula (18.2%). Binary regression analysis coefficient (B) revealed that the rate of patients acquiring tolerance at the end of the study was influenced by 2 factors: (1) IgE-mediated mechanism (B −2.05, OR 0.12, 95% CI 0.06-0.26; P < .001); and (2) formula choice, such that those receiving either EHCF (B 1.48, OR 4.41, 95% CI 1.44-13.48; P = .009) or EHCF + LGG (B 3.35, OR 28.62, 95% CI 8.72-93.93; P < .001). Conclusions EHCF accelerates tolerance acquisition in children with CMA if compared with other dietetic choices. This effect is augmented by LGG.
Milk coagulation properties (MCP) are conventionally measured using computerized renneting meters, mechanical or optical devices that record curd firmness over time (CFt). The traditional MCP are ...rennet coagulation time (RCT, min), curd firmness (a30, mm), and curd-firming time (k20, min). The milk of different ruminant species varies in terms of CFt pattern. Milk from Holstein-Friesian and some Scandinavian cattle breeds yields higher proportions of noncoagulating samples, samples with longer RCT and lower a30, and samples for which k20 is not estimable, than does milk from Brown Swiss, Simmental, and other local Alpine breeds. The amount, proportion, and genetic variants (especially κ-casein) of milk protein fractions strongly influence MCP and explain variable proportions of the observed differences among breeds and among individuals of the same breed. In addition, other major genes have been shown to affect MCP. Individual repeatability of MCP is high, whereas any herd effect is low; thus, the improvement of MCP should be based principally on selection. Exploitable additive genetic variation in MCP exists and has been assessed using different breeds in various countries. Several models have been formulated that either handle noncoagulating samples or not. The heritability of MCP is similar to that of other milk quality traits and is higher than the heritability of milk yield. Rennet coagulation time and a30 are highly correlated, both phenotypically and genetically. This means that the use of a30 data does not add valuable information to that obtainable from RCT; both traits are genetically correlated mainly with milk acidity. Moreover, a30 is correlated with casein content. The major limitations of traditional MCP can be overcome by prolonging the observation period and by using a novel CFt modeling, which uses all available information provided by computerized renneting meters and allows the estimation of RCT, the potential asymptotic curd firmness, the curd-firming rate, and the syneresis rate. Direct measurements of RCT obtained from both mechanical and optical devices show similar heritabilities and exhibit high phenotypic and genetic correlations. Moreover, mid-infrared reflectance spectroscopy can predict MCP. The heritabilities of predicted MCP are higher than those of measured MCP, and the 2 sets of values are strongly correlated. Therefore, mid-infrared reflectance spectroscopy is a reliable and cheap method whereby MCP can be improved at the population level; this is because such spectra are already routinely acquired from the milk of cows enrolled in milk recording schemes.
Critical advancement is needed in the study of human milk as a biological system that intersects and interacts with myriad internal (maternal biology) and external (diet, environment, infections) ...factors and its plethora of influences on the developing infant. Human-milk composition and its resulting biological function is more than the sum of its parts. Our failure to fully understand this biology in a large part contributes to why the duration of exclusive breastfeeding remains an unsettled science (if not policy). Our current understanding of human-milk composition and its individual components and their functions fails to fully recognize the importance of the chronobiology and systems biology of human milk in the context of milk synthesis, optimal timing and duration of feeding, and period of lactation. The overly simplistic, but common, approach to analyzing single, mostly nutritive components of human milk is insufficient to understand the contribution of either individual components or the matrix within which they exist to both maternal and child health. There is a need for a shift in the conceptual approach to studying human milk to improve strategies and interventions to support better lactation, breastfeeding, and the full range of infant feeding practices, particularly for women and infants living in undernourished and infectious environments. Recent technological advances have led to a rising movement towards advancing the science of human-milk biology. Herein, we describe the rationale and critical need for unveiling the multifunctionality of the various nutritional, nonnutritional, immune, and biological signaling pathways of the components in human milk that drive system development and maturation, growth, and development in the very early postnatal period of life. We provide a vision and conceptual framework for a research strategy and agenda to change the field of human-milk biology with implications for global policy, innovation, and interventions.
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Cow’s milk protein allergy (CMPA) may cause gastrointestinal motility disorders. Symptoms of both conditions overlap and diagnostic tests do not reliably differentiate between both. A decrease of ...symptoms with an extensive hydrolysate and relapse during challenge is not a proof of allergy, because hydrolysates enhance gastric emptying, a pathophysiologic mechanism of gastro‐oesophageal reflux (GER). Thickened formula reduces regurgitation, and failure to do so suggests CMPA. A thickened extensive hydrolysate may induce more rapid improvement, but does not always differentiate between CMPA and GER. Different hypotheses are discussed: is the overlap between CMPA and functional disorders coincidence, or do both entities present with identical symptoms, or does the fact that symptoms are identical indicates that there is only one entity involved? Studies on the prevention of CMPA focused on ‘at‐risk families’, and resulted in a decrease of CMPA and atopic dermatitis, but did not provide data on the incidence of GER.
Conclusion: As long as there are no objective diagnostic tools to separate GER from CMPA, the physician has two options: first treat the most likely diagnosis, and switch if after 2–4 weeks there is no improvement, or treat both conditions with one intervention, what will not result in a diagnosis.
Nutraceutical and functional food market are one of the fastest-growing food segments in the newer food product development category. In the recent past, focus of beverage industry has shifted ...towards making food more nutritious and functionally enriched. To load the food with nutraceutical functionality, herbal sources having target functional compounds are either directly used as food or for separation of target compounds. Among beverages, milk is considered as a wholesome complete food providing macro (fat, proteins, and carbohydrates) and micronutrients (calcium, selenium, riboflavin, vitamin B
12
, and pantothenic acid vitamin B
5
) in balanced proportions. However, limited access to milk in some regions of globe, low availability of certain minerals (iron), vitamins (folate), and other biomolecules (amino acids) compounded with issues like milk allergy, lactose intolerance, and hypercholesterolemia have forced some specific population groups to search for better milk alternatives which are more or at least equi-nutritional to conventional milk. Plant-based wholesome or blended milk analogs are better studied as inexpensive alternates to conventional milk for people who are in search of better alternates for one or other reason. The market of milk analogs is currently dominated by soya bean milk, oat milk, coconut milk, hemp milk, cocoa milk, multigrain milk etc. most of which are produced by controlled fermentation which owes to their functional bioactive composition. Such analogs are appreciated for their functionally active components which are often correlated to their health-promoting and disease-preventing properties. One major advantage of analogs over conventional milk is that the energy input per unit of milk produced is much less compared to animal milk while there is always an opportunity to manipulate their composition based on demand. However, the major limiting factor in the acceptance of such non-conventional beverages is their challenging production technology and poor sensory profile which is true particularly for beverages derived from legumes. These challenges provide an opportunity for both industries and research personals to put in major concerted efforts in the field of functional bioactive food segment to produce tailor-made novel beverages which are nutritional, economic and have improved functionality. Keeping in view the potential of plant-based milk alternates and associated challenges the aim of the present review is to give a scientifically comparative and conclusive overview of the present status, market potential, and health concerns of plant-based nondairy milk beverages.
An allergy to cow's milk requires the avoidance of cow's milk proteins and, in some infants, the use of a hypoallergenic formula. This review aims to summarize the current evidence concerning ...different types of hydrolysed formulas (HF), and recommendations for the treatment of IgE- and non-IgE-mediated cow's milk allergy and functional gastrointestinal disorders in infancy, for which some dietary intervention and HF may be of benefit to both immune and motor mechanisms. Current guidelines recommend cow's milk protein (i.e., whey or casein) extensively hydrolysed formula (eHF) as the first choice for cow's milk allergy treatment, and amino acid formulas for more severe cases or those with reactions to eHF. Rice hydrolysed formulas (rHF) have also become available in recent years. Both eHF and rHF are well tolerated by the majority of children allergic to cow's milk, with no concerns regarding body growth or adverse effects. Some hydrolysates may have a pro-active effect in modulating the immune system due to the presence of small peptides and additional components, like biotics. Despite encouraging results on tolerance acquisition, evidence is still not conclusive, thus hampering our ability to draw firm conclusions. In clinical practice, the choice of hypoallergenic formula should be based on the infant's age, the severity, frequency and persistence of symptoms, immune phenotype, growth pattern, formula cost, and in vivo proof of tolerance and efficacy.
The objectives were to evaluate the effects of elevated pre- and postpartum nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHBA) concentrations during the transition period on reproductive ...performance and milk production in dairy cattle. In a prospective cohort study of 91 freestall, total mixed ration-fed herds in the northeastern United States, blood samples were collected from approximately 15 prepartum and 15 different postpartum transition animals in each herd. All samples were stratified based on pre- or postpartum status at the time of sample collection, and 2,259 and 2,290 animals were used to evaluate reproductive and milk production performance, respectively. Reproductive performance was assessed by time to conception within 70 d post-voluntary waiting period (VWP) and milk production was assessed using mature-equivalent 305-d (ME305) milk yield estimated at 120 d in milk. While controlling for body condition score (BCS), calving season, median ME305 milk production, and parity, NEFA and BHBA concentrations were evaluated with time to event analysis to investigate reproductive performance. These same predictor variables were used to determine the effects of elevated NEFA and BHBA concentrations on ME305 milk yield with herd as a random effect. Heifers and cows were grouped in the final analyses if the results between groups were similar. In all animals sampled prepartum, the risk of pregnancy within 70 d post-VWP was reduced by 19% when NEFA concentrations were ≥0.27mEq/L. In all animals sampled postpartum, those with NEFA concentrations ≥0.72mEq/L had a 16% decrease in risk of pregnancy and those with BHBA concentrations ≥10mg/dL had a 13% decrease in risk. In cows and heifers, ME305 milk yield was decreased by 683kg when prepartum NEFA concentrations were ≥0.33mEq/L. In heifers sampled postpartum, ME305 milk yield was increased by 488kg when NEFA concentrations were ≥0.57mEq/L and increased by 403kg when BHBA concentrations were ≥9mg/dL. In cows sampled postpartum, ME305 milk yield was decreased by 647kg when NEFA concentrations were ≥0.72mEq/L and decreased by 393kg when BHBA concentrations were ≥10mg/dL. With the exception of milk production in heifers, this study indicates that increased concentrations of serum NEFA and BHBA had a detrimental effect on reproductive performance and milk production.
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