Serum substitute in epithelial cell culture media: nonfat dry milk filtrate

A number of milk types and milk fractions were investigated as possible substitutes for serum in cell culture media. A filtrate of reconstituted nonfat dry milk showed promise. Culture fluids containing 5% of the nonfat dry milk filtrate were used to propagate primary and continuous cell cultures, and the cell growth from these cultures was compared with that of cells grown in a serum-containing medium. The nonfat dry milk filtrate-supplemented medium supported the growth of all epithelial cells tested, but two fibroblast-type cultures failed to replicate. Cells grown in the medium containing the milk filtrate grew slowly for 2 to 3 days and then propagated to confluency in 6 to 8 days. Viable cell counts of 9 days were comparable to those of serum-grown cells that had been propagated for 7 days. Cells grown in the milk filtrate could be split 1 to 4 when subcultures were prepared. Cell growth could be stimulated by refeeding on days 2 to 3 or by the addition of 30 microM 2-mercaptoethanol to the growth medium. Virus susceptibility and titer comparisons with poliovirus 1, coxsackievirus B2, echovirus 7, and herpes simplex virus indicated that approximately the same data were obtained when either the nonfat dry milk filtrate-treated or the serum-treated cells were studied. The nonfat dry milk filtrate is inexpensive, is easily prepared, and is a substitute for serum in epithelial cell culture media.     

Serum substitute in epithelial cell culture media: nonfat dry milk filtrate.

A number of milk types and milk fractions were investigated as possible substitutes for serum in cell culture media. A filtrate of reconstituted nonfat dry milk showed promise. Culture fluids containing 5% of the nonfat dry milk filtrate were used to propagate primary and continuous cell cultures, and the cell growth from these cultures was compared with that of cells grown in a serum-containing medium. The nonfat dry milk filtrate-supplemented medium supported the growth of all epithelial cells tested, but two fibroblast-type cultures failed to replicate. Cells grown in the medium containing the milk filtrate grew slowly for 2 to 3 days and then propagated to confluency in 6 to 8 days. Viable cell counts of 9 days were comparable to those of serum-grown cells that had been propagated for 7 days. Cells grown in the milk filtrate could be split 1 to 4 when subcultures were prepared. Cell growth could be stimulated by refeeding on days 2 to 3 or by the addition of 30 microM 2-mercaptoethanol to the growth medium. Virus susceptibility and titer comparisons with poliovirus 1, coxsackievirus B2, echovirus 7, and herpes simplex virus indicated that approximately the same data were obtained when either the nonfat dry milk filtrate-treated or the serum-treated cells were studied. The nonfat dry milk filtrate is inexpensive, is easily prepared, and is a substitute for serum in epithelial cell culture media.     

Performance of calves fed on milk replacers containing fish protein hydrolysate

Milk substitutes for calves are based upon dried skim milk and added fats of vegetable or animal origin. Many attempts have been made to replace part of the skim milk in these feeds by cheaper sources of protein.A milk product coagulates in the calf's abomasum and has a high digestibility. Milk replacers based on fish protein hydrolysate (FPH) do not coagulate in the abomasum, but have a high apparent digestibility coefficient (0.92) and the amino acid balance is similar to that of skim milk.Calves were fed on milk replacers prepared from FPH made from various materials. In general, FPH prepared from white fish (cod, blue whiting, white fish offal) was satisfactory although calf performance until weaning was poorer than for calves fed on milk replacers with dried skim milk (SMP). Some ways are suggested by which the quality of the product (FPH) could be maintained. FPH preparations from fatty fish, like sprats and mackerel, were also used. An antioxidant was added in the preparation of sprats and mackerel because of the unsaturated nature of fish fats. Milk replacers containing sprats prepared without an antioxidant and mackerel (31% body fat) with antioxidant were unsatisfactory and further development work is required on FPH materials from fatty fish.FPH from white fish can be dried without any appreciable loss in calf performance. The daily gain to weaning was 0.25 and 0.29 kg per day for calves fed on the dried and undried material, respectively. On the basis of present knowledge it is suggested that FPH can replace one third of skim milk in milk replacers for early weaned calves; the proportion recommended may rapidly increase with further technical development of the product.     

Evaluation of two milk replacers fed to hand‐reared cheetah cubs (Acinonyx jubatus): nutrient composition,apparent total tract digestibility,and comparison to maternal cheetah milk

Commercially prepared milk replacers are frequently used to provide the sole source of nutrition for hand‐reared cheetah cubs (Acinonyx jubatus). The nutrient composition of two commonly used milk replacers was determined. Using titanium dioxide as an indigestible marker, nutrient digestibility was calculated from the analyses of fecal samples collected from each cub (n = 4 on formula 1, and n = 2 on formula 2). Mean apparent total tract digestibility for both formulas was >90% for all nutrients analyzed (crude protein, amino acids, crude fat (CF), and dry matter). However, the total CF content and the concentration of the essential fatty acids, such as α‐linolenic, linolenic, and arachidonic acid, of both formulas was lower than reported for maternal cheetah milk. Additionally, one formula contained a comparatively high amount of carbohydrate, at the expense of protein. Although data were lacking for cheetah maternal milk, comparison with domestic cat milk revealed high concentrations of a number of minerals (K, Fe, Zn, and Cu), while vitamin D₃ was not detected in one formula. Both formulas were low in the majority of essential amino acids compared with domestic cat maternal milk. Despite their apparently high digestibility, neither formula was complete or balanced in terms of nutrient concentrations and ratios when maternal cheetah milk and/or the requirements established for growth in domestic cats were used as estimates of ideal. On this basis, although all cubs in this study were healthy and maintained good body conditions for the duration of the trial, the results of dietary analyses indicate that these milk replacers may not provide optimal nutrition for growth in cheetah cubs when used for extended periods. Zoo Biol 30:412–426, 2011. © 2010 Wiley‐Liss, Inc.     

Use of fish protein hydrolysate in milk replacers

Fish protein hydrolysate (FPH) has been used as the sole source of protein in milk replacers for lambs in several experiments. In comparison with casein, diets containing FPH gave slightly inferior growth rates during the first 2 weeks of a 5-week rearing period. The lower growth rate was generally compensated for in the following 3 weeks so that overall growth rate and food utilization were similar with milk replacers containing casein or FPH. Diets based on FPH, lard and hydrolysed starch were found to give results similar to a milk replacer based on casein, butterfat and lactose. The use of fat fish to provide both the source of fat and protein has received some attention and needs to be further investigated.     

Development of a fluorogenic probe-based PCR assay for detection of Bacillus cereus in nonfat dry milk

A fluorogenic probe-based PCR assay was developed and evaluated for its utility in detecting Bacillus cereus in nonfat dry milk. Regions of the hemolysin and cereolysin AB genes from an initial group of two B. cereus isolates and two Bacillus thuringiensis isolates were cloned and sequenced. Three single-base differences in two B. cereus strains were identified in the cereolysin AB gene at nucleotides 866, 875, and 1287, while there were no species-consistent differences found in the hemolysin gene. A fluorogenic probe-based PCR assay was developed which utilizes the 5'-to-3' exonuclease of Taq polymerase, and two fluorogenic probes were evaluated. One fluorogenic probe (cerTAQ-1) was designed to be specific for the nucleotide differences at bases 866 and 875 found in B. cereus. A total of 51 out of 72 B. cereus strains tested positive with the cerTAQ-1 probe, while only 1 out of 5 B. thuringiensis strains tested positive. Sequence analysis of the negative B. cereus strains revealed additional polymorphism found in the cereolysin probe target. A second probe (cerTAQ-2) was designed to account for additional polymorphic sequences found in the cerTAQ-1-negative B. cereus strains. A total of 35 out of 39 B. cereus strains tested positive (including 10 of 14 previously negative strains) with cerTAQ-2, although the assay readout was uniformly lower with this probe than with cerTAQ-1. A PCR assay using cerTAQ-1 was able to detect approximately 58 B. cereus CFU in 1 g of artificially contaminated nonfat dry milk. Forty-three nonfat dry milk samples were tested for the presence of B. cereus with the most-probable-number technique and the fluorogenic PCR assay. Twelve of the 43 samples were contaminated with B. cereus at levels greater than or equal to 43 CFU/g, and all 12 of these samples tested positive with the fluorogenic PCR assay. Of the remaining 31 samples, 12 were B. cereus negative and 19 were contaminated with B. cereus at levels ranging from 3 to 9 CFU/g. All 31 of these samples were negative in the fluorogenic PCR assay. Although not totally inclusive, the PCR-based assay with cerTAQ-1 is able to specifically detect B. cereus in nonfat dry milk.     

Artificial diets used in mass production of the New World screwworm,Cochliomyia hominivorax

The New World screwworm, Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae), has been eradicated from North and Central America using the sterile insect technique. This success has been based on mass production of high‐quality screwworms using artificial diets since 1958. Many diet formulations for both larvae and adults have been developed, mainly driven by cost efficiency and material supply. However, only four larval and two adult diet formulations have been applied in the six sterile fly mass production plants in the USA, Mexico and Panama. Herein, we briefly review the history of screwworm diet research and development, introduce the diet formulations used in mass rearing and discuss their advantages and disadvantages in terms of plant application. Finally, we propose future research on screwworm nutrition, potential protein sources, feeding stimulants, further optimization of screwworm formulations and possible methods to reduce the negative qualities of waste generated during the mass production.     

Use of nonfat dry milk to block nonspecific nuclear and membrane staining by avidin conjugates

The use of fluorescein-avidin or rhodamine-avidin conjugates in conjunction with biotinylated secondary antibodies for indirect immunohistology frequently results in pronounced nonspecific nuclear staining in kidney sections. This nonspecific nuclear staining can be effectively blocked by using 5% (w/v) nonfat dry milk in buffered saline as the diluent for the avidin conjugates. In contrast, 5% (w/v) bovine serum albumin, a commonly used blocking agent, has only a modest effect. Nonfat dry milk is also effective as a blocking agent and carrier when used in fluorescence-activated cell sorter (FACS) analysis. These results emphasize the broad usefulness of milk-based blocking reagents.     

Assessment of the production costs of fish protein hydrolysates

An assessment of the production costs of fish protein hydrolysates has shown that, while costs will be higher than for fish meal production, a more detailed economic examination would be warranted. The estimated cost of the dried product suggests that hydrolysates, high in protein, will be attractive as milk replacers.     

Differentiation between carcasses from suckling lambs reared with ewe milk or milk replacers by near infrared reflectance spectroscopy of perirenal fat

Near infrared reflectance spectroscopy (NIRS) was used to discriminate between carcasses from Churra breed suckling lambs reared with ewe milk or milk replacers. Samples were scanned over the NIR spectral range (1100–2500 nm). The results showed that NIRS could be used successfully to discriminate the suckling lambs depending on milk source, with a 100% of correctly classified samples. NIRS technology would be a good method, since it is a rapid, economical and little complex method.     

Indigestion in Young Calves

Increasing the lactose content of different milk replacers or milk diets by approx. 30 % of the dry matter increased the frequency of diarrhoea the first 10–12 days in young calves on all occasions in 5 experiments comprising 120 calves. For all diets taken together, this effect was highly significant. Total daily intakes of lactose amounted to 200–480 g. When lactose was given on top of the milk rations, the growth rate increased significantly, whereas the growth rate was usually insignificantly reduced when lactose replaced other nutrients in milk diets or milk replacers, their levels of protein and fat becoming low. Albumin and total protein in blood plasma were significantly lower when the dietary protein level was low. Milk replacers with 20 or 40 % whey powder, replacing skim milk powder, performed equally well, but gave significantly less growth than the old-fashioned feeding of whole milk-skim milk. Intake of hay and barley and a number of clinical and histological or pathological parameters did not vary consistently with dietary level of whey powder or lactose. Feeding whole milk all the time resulted in low intake of hay and barley and poorly developed forestomachs, but high dressing-out %. Substituting soya for part of the skim milk powder in milk replacers gave abomasal content with no curds. In most cases, pH in the rumen appeared to be nearly up to neutral until the calves ate ground barley, about 1 month old.     

The incorporation of solubilized wheat proteins in milk replacers for veal calves: effects on growth performance and muscle oxidative capacity

Replacement of skim milk proteins by solubilized wheat protein (SWP) in milk replacers for veal calves would contribute to the reduction in feeding costs. The occurrence of metabolic disorders has, however, been reported. Forty-two male calves received one of three treatments over 140 days: a control diet, a diet containing SWP without or with branched-chain amino acid supplementation. Liveweight gain, carcass yield, color and conformation did not show any significant differences. No metabolic disorders were noted. Supplementation with branched-chain amino acids reduced the marginal Val deficiency but did not modify the growth performances. With the SWP containing diets, the plasma metabolite profile was characteristic of those observed with non-clotting diets. It was statistically correlated to the changes in the orientation of the Semitendinosus muscle energy metabolism towards a more oxidative type and to indications of a lower efficiency of amino acid utilisation for protein deposition. At the present levels of inclusion, SWP proved to be an interesting alternative to the sole use of whey as the protein source in milk replacers for veal calves.     

Danish Holsteins Favor Bull Offspring: Biased Milk Production as a Function of Fetal Sex,and Calving Difficulty

In a previous study from 2014 it was found that US Holstein cows that gave birth to heifer calves produced more milk than cows having bull calves. We wanted to assess whether this is also true for Danish cattle. Data from 578 Danish Holstein herds were analysed with a mixed effect model and contrary to the findings in the US, we found that cows produced higher volumes of milk if they had a bull calf compared to a heifer calf. We found a significantly higher milk production of 0.28% in the first lactation period for cows giving birth to a bull calf, compared to a heifer calf. This difference was even higher when cows gave birth to another bull calf, so having two bull calves resulted in a difference of 0.52% in milk production compared to any other combination of sex of the offspring. Furthermore, we found that farmer assisted calvings were associated with a higher milk yield. Cows with no farmer assistance or with veterinary assistance during the most recent calving produced less milk. There were also indications that dams would favor a bull fetus by decreasing milk production during the second pregnancy if the calf born in the first parity was a heifer. We hypothesize that size of calves is a confounding factor for milk production. However, calving weight was not available in the present data set to test this hypothesis.     

Humanizing infant milk formula to decrease postnatal HIV transmission

There are currently no safe methods for feeding babies born from the 16 million HIV-infected women living in resource-constrained countries. Breast milk can transmit HIV, and formula feeding can lead to gastrointestinal illnesses owing to unsanitary conditions and the composition of milk formulations. There is therefore a need to ensure that breast milk substitutes provide optimal health outcomes. Given that the immune properties of several breast milk proteins are known, transgenic food crops could facilitate inexpensive and safe reconstitution of the beneficial breast milk proteome in infant formulae, while keeping the HIV virus at bay. At least seven breast milk immune proteins have already been produced in food crops, and dozens more proteins could potentially be produced if fortified formula proves effective in nursing newborns born to HIV-infected mothers.     

The replacement of skim milk by dried microbial cells and whey in milk diets for calves

In two experiments, the performance and health of calves fed on diets containing dried microbial cells (DMC) was monitored. In the first experiment 32 seven-day-old British Friesian bull calves, reared on a conventional early weaning system to 8 weeks of age, were given milk replacers in which 5, 10 and 20% spray-dried DMC and whey replaced 14–55% of skim milk protein. All DMC diets contained 36 g/kg of non-nucleic acid nitrogen. In Experiment 2, 38 two-day-old Ayrshire bull calves were fed for 63 days solely on one of two milk diets containing 0 or 10% DMC at 5 levels of intake designed to produce body gains of 0–1 kg/day. While the inclusion of up to 10% DMC in both experiments had little effect on performance or health, 20% DMC was associated with severe diarrhoea, loss of appetite and growth retardation, and impaired clotting of the milk in vitro. In Experiment 2 the response, in gain, by calves given increasing intakes of milk dry matter was linear and was unaffected by the type of diet offered.It was concluded that when levels of DMC in excess of 10% inclusion are used in association with maize starch, deleterious effects on performance are likely.     

Subspecies and body size allometry affect milk production and composition, and calf growth in red deer: comparison of Cervus elaphus hispanicus and Cervus elaphus scoticus

Studies comparing lactation in wild mammals have shown that maternal weight scales with offspring weight, milk production, or its energy. However, no study appears to have scaled milk composition with maternal or offspring weight. Although diet affects milk composition and production, their effects in biological studies have almost never seemed to be controlled. In this study, we compare two subspecies of red deer, Scottish deer, Cervus elaphus scoticus (10 lactations), and Iberian deer, Cervus elaphus hispanicus (14 lactations), kept under the same diet and housing to assess differences in hind and calf weights and their trends, milk production and composition, and their allometric relationships. Scottish hinds were heavier, and calf weight and gains were greater than Iberian ones, with greater milk production and milk protein content, but they did not differ in fat or lactose content. Calf birth weight, milk production, and protein content showed significant allometric relationships with maternal weight, but no relationship was found for fat, lactose, or any of these variables with calf birth weight. Protein content correlated with calf birth weight, and calf weight trend depended on milk protein production rather than on that of fat or lactose. Protein may be the most important milk component to explain growth and milk composition differences between closely related mammals.     

Influence of 48 hour calf separation on milk production and calf growth in range cows

Hereford cows and their calves were either left together or separated for a 48-hr period between 50 and 80 days postpartum. Milk production and calf weights were determined 1 and 2 weeks prior to and 1 and 3 weeks after calf separation. Daily milk production of separated cows (5.6+/-.1 kg) was not different from that of control cows (5.3+/-.1 kg) at any sampling period. Similarly, calf growth was not affected (P > .10) by separation; both groups of calves gained .64 kg/day. Average 205-day adjusted weaning weights were also similar, for the control (173.5+/-4.6 kg) and separated calves (181.8+/-4.6 kg). These results indicate that 48-hr calf separation could be used in a treatment regime to decrease the postpartum anestrous interval in range cattle without detrimental effects on milk production, calf growth or 205-day adjusted weaning weight.     

Effect of Zinc Methionine or Zinc Sulfate Supplementation on Milk Production and Composition of Milk in Lactating Dairy Cows

Eighteen lactating dairy cows were used to compare the effects of organic and inorganic Zn supplements on milk production and chemical composition of milk. Animals received three diets in a randomized block design: basal diet with no supplemental Zn (control, 42 mg Zn/kg), basal diet plus 500 mg Zn/kg of dry matter (DM) as zinc sulfate monohydrate (ZnS) and basal diet plus 500 mg Zn/kg of DM as zinc methionine (ZnM). Results showed that milk and fat-corrected milk yield in dairy cows were not significantly affected by Zn source although a numerical increase was observed. The percentages of protein, lactose, fat, solid nonfat, total solid, and density of milk were not significantly different between treatments. However, dairy cows that received ZnM tended to produce more milk and fat-corrected milk with a lower somatic cell count as compared to controls. The zinc concentration in milk in the ZnM and ZnS groups was higher (P < 0.05) than in milk from the control group, but there were no significant differences between ZnS and ZnM groups.     

Influence of the dam and sire on size at birth and subsequent growth, milk production and fertility in dairy heifers

Genetic selection has resulted in larger cows with high milk production potential but a tendency for poor fertility. In multiparous cows fetal development competes for nutrients with concurrent milk production. This study tested the hypotheses that (a) maternal age and milk yield during pregnancy alter calf birth size and (b) birth weight influences subsequent productivity and fertility. Concurrently born Holstein-Friesian heifers (n=65) with multiparous dams and three sires were monitored from birth to the end of their first lactation to assess effects of birth weight on growth, milk production and fertility. Calves were analyzed as three subgroups: low (L), average (A) and high (H) birth weight (BW) calves (n=21-22 per group). LBW calves were born 10 kg lighter than HBW calves and remained significantly lighter throughout the study. They were generally smaller in other measured indices (length, height, girth, ponderal index) between birth and 9 months and were more likely to have older dams (lactations 3-6) with higher peak yields (>42 kg/day). Milk production parameters were indistinguishable between the 3 birthweight groups and metabolic parameters (IGF-I, insulin, glucose) measured around first calving were unaffected. HBW offspring were more likely to have persistent corpora lutea following their first calving and other fertility parameters also tended to be worse. Sire influenced gestation length but not birth size. Sire heritability estimates showed that weight, IGF-I and insulin concentrations after first calving and fertility in the first lactation were all heritable. The results support the hypothesis that high milk production in the dam may predispose to birth of a smaller calf. Smaller birth size did not, however, have any subsequent adverse effects on productivity or fertility in the first lactation and sire was more influential at this stage.     

Neutralization of the Bactericidal Effect of Cocoa Powder on Salmonellae by Casein

Four pre-enrichment media (nutrient broth, lactose broth, reconstituted non-fat dry milk and nutrient broth containing 5% (w/v) casein) were evaluated for the recovery of salmonellae from cocoa powder. Addition of 5% cocoa powder to nutrient broth and to lactose broth proved to be bactericidal to salmonellae. Heat-damaged Salmonella typhimurium were more sensitive than undamaged cells to cocoa powder and agitation increased the bactericidal effect. The bactericidal effects were minimized in pre-enrichment media that contained either 5% casein or nonfat dry milk.