• Asian-Australasian Journal of Animal Sciences
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• v.15 no.6
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• pp.895-899
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• 2002

Lysozyme activity in buffalo milk in relation to the period of lactation, parity of animal, weather conditions and udder infections was studied. Effect of storage and heat processing of milk on lysozyme activity was determined. Lysozyme activity was higher in buffalo milk than in cow milk. Buffalo colostrum showed lysozyme activity 5 times of that in mature milk. Lysozyme activity in buffalo milk was not influenced by the parity of animal and the stage of lactation, however, it increased during extreme whether conditions (winter and summer). Lysozyme in both cow and buffalo milk exhibited maximum activity at pH 7.4. Buffalo milk lysozyme was fully stable while the cow milk lysozyme was partly inactivated by pasteurization (low temperature-long time as well as high temperature-short time treatments). Lysozyme in buffalo milk was more stable than in cow milk during storage and heat treatment. A 10 to 50-fold increase in milk lysozyme activity was observed in mastitic cows. An assay of lysozyme activity in milk can be used to diagnose mastitis in cattle but not in buffaloes. Some buffaloes exhibited 1000 fold greater lysozyme activity and moderately raised somatic cell count in milk, but there was no sign of mastitis in these animals. A possible role of milk lysozyme in prevention of mastitis in buffaloes is discussed.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.12
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• pp.1774-1783
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• 2015

Milk lysozyme is the ubiquitous enzyme in milk of mammals. In this study, the cDNA sequence of a new chicken-type (c-type) milk lysozyme gene (YML), was cloned from yak mammary gland tissue. A 444 bp open reading frames, which encodes 148 amino acids (16.54 kDa) with a signal peptide of 18 amino acids, was sequenced. Further analysis indicated that the nucleic acid and amino acid sequences identities between yak and cow milk lysozyme were 89.04% and 80.41%, respectively. Recombinant yak milk lysozyme (rYML) was produced by Escherichia coli BL21 and Pichia pastoris X33. The highest lysozyme activity was detected for heterologous protein rYML5 (M = 1,864.24 U/mg, SD = 25.75) which was expressed in P. pastoris with expression vector $pPICZ{\alpha}A$ and it clearly inhibited growth of Staphylococcus aureus. Result of the YML gene expression using quantitative polymerase chain reaction showed that the YML gene was up-regulated to maximum at 30 day postpartum, that is, comparatively high YML can be found in initial milk production. The phylogenetic tree indicated that the amino acid sequence was similar to cow kidney lysozyme, which implied that the YML may have diverged from a different ancestor gene such as cow mammary glands. In our study, we suggest that YML be a new c-type lysozyme expressed in yak mammary glands that plays a role as host immunity.

• The Journal of Natural Sciences
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• v.7
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• pp.53-58
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• 1995

To elucidate the effect of egg white lysozyme from Ogol fowl on the preservation of milks, fishcurd and sausage, changes of pH, volatile base nitrogen content and viable cell count were investigated during the storage periods at $20^{\circ}C$, $30^{\circ}C$ and $37^{\circ}C$ after the addition of lysozyme in each foods. Volatile base nitrogen count of raw milk as marker of spoilage was lowest(63 mg%) in 0.05% lysozyme addition lot which was stored at $20^{\circ}C$ for 2 days, and its preservation effect by lysozyme at $30^{\circ}C$ was enhanced with addition of glycine(0.1%). Preservation effect by lysozyme in commercial milk at $37^{\circ}C$ and in fishcurd at $5^{\circ}C$ and $20^{\circ}C$ were also good, and when sausage was stored at $5^{\circ}C$ after treatment of lysozyme instead of sorbic acid, its preservation effect was acceptable.

• Food Science of Animal Resources
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• v.43 no.4
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• pp.674-684
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• 2023

Xinjiang province is the main camel feeding area in China with a large square, and camel milk from different areas have different qualities. By now, there are few reports about the quality of camel milk from different areas of Xinjiang province in China. In this study, seven batches of camel milk and one batch of cow milk were collected, and the contents of fat, protein, lactose, total solid, and nonfat milk solid of these milk samples were determined, as well as the contents of lysozyme and vitamin C. All samples were scored and compared by principal component analysis score and comprehensive weighted multi-index score. As the results, camel milk from different areas showed different contents of fat (4.62%-7.02%), protein (3.34%-3.95%), lactose (3.85%-4.79%), total solid (13.59%-17.00%), nonfat milk solid (8.55%-9.73%), vitamin C (12.10-41.25 ㎍/mL), and lysozyme (8.70-22.80 ㎍/mL), as well as different qualities. This variation would help people to know more about quanlity of camel milk in Xinjiang province. Camel milk from Jeminay showed the best quality, and then followed by camel milk from Fukang, Changji, and Fuhai, while cow milk showed the lowest score. Therefore, Jeminay is the most suitable place for grazing camels. Our findings show the different qualities of camel milk in different distribution areas of Xinjiang province, and provide an insight for the evaluation of camel milk. In the present study, only seven components in camel milk were determined, many other factors, such as cfu, mineral, and other vitamins, have not been considered.

• Journal of Nutrition and Health
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• v.37 no.9
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• pp.809-816
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• 2004

The purpose of this study was to evaluate the effect of maternal nutritional status and health behaviors on the concentrations of minerals (Zn, Fe, Ca) and the immunological substances (lactoferrin, sIgA, Iysozyme) in breast milk. Breast milk was collected from 193 healthy Korean women from obstetric clinics and postpartum care centers in Seoul. : 99 colostrum (1 - 5 days postpartum), 33 transitional milk (6 - 10 days postpartum), 61 mature milk (11 - 50 days postpartum). The concentrations of minerals and immunological substance were highest in colostrum and decreased with lactational period. Concentrations of Zn and Fe reduced significantly from colostrum to mature milk, however, Ca concentration stayed constant throughout the lactational period. Contents of lactoferrin, sIgA, and lysozyme were significantly lower in mature milk than in colostrum. Mother's nutritional status, assessed by prepregnancy BMI, had an effect only on colostrum, but not on transition and mature milk. Fe concentration of colostrum was significantly lower in underweight (prepregnancy BMI < 18.5) than in overweight mothers (prepregnancy BMI $\geq$ 23.0). Also lower tendency was observed for sIgA and lysozyme contents, even though the difference was not statistically significant. Pregnancy weight gain had no effect on the breast milk component. Since nutritional factors had some effect on colostrum, the health behaviors of mothers providing colostrum were assessed. The mother's behavior of smoking, drinking, morning sickness, parity, disease, nutrient supplement use had no significant effect on the breast milk component, however, Zn, sIgA, and lysozyme were the somewhat affected components by maternal health behavior.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.4
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• pp.462-466
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• 2001

To understand molecular mechanisms of mouse mammary gland involution, clones were isolated by differential screening of a cDNA library. Partial sequences of a clone showed 100% identity to cDNA sequences of mouse lysozyme P gene. Northern analysis was performed to examine expression levels of lysozyme mRNA in mammary gland at several physiological states. Expression of lysozyme gene was induced at involution day 5 compared with lactating stage. High levels of lysozyme mRNA were also detected at virgin tissues. Two types of separate genes, P and M lysozyme, have been known in mouse, and we found that both lysozyme P and M genes were expressed in mammary tissues by reverse transcriptase-polymerase chain reaction. The lysozyme enzyme activity determined by lysoplate assay was also higher in involuted mammary tissues compared with lactating tissues, showing a similar trend to its mRNA levels. Lysozyme is an antimicrobial protein and involved in host defense mechanism. The increase in lysozyme gene expression may help to prevent microbial infection during mammary gland involution at which stage the residual milk in the mammary gland provides good nutritional sources for microbial growth.

• Journal of Nutrition and Health
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• v.29 no.6
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• pp.569-577
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• 1996

This study was performed to determine the effect of maternal protein intake on 1) the concentration of immune substances in milk 2) degree of passive immunity to pups via lactation, and 3) specific antibody production to a specific antigen, $\beta$-lactoglobulin(BLG). 4) the effect of passive immunity that pups received from mother during lactation on the production of antibodies when the pups were challenged to the same antigen. Part of the female rats were immunized with BLG before and during pregnancy. The pregnant rats were placed into either 25% or 10% isolated soy protein diet throughout gestation and lactation. After weaning, pups from each group continued to be fed the same diet. At 18 weeks of age, all the pups were challenged with BLG. Total IgA and IgG, lysozyme, BLG-specific IgA and IgG were measured in dam's serum, dam's milk, and pup's serum. Total IgG, and lysozyme in dam's serum and milk were higher in high protein group. Total IgA and IgG in pup's serum remained higher in high protein group from 5 to 18 weeks of age. BLG-specific antibodies were found in the milk and serum of immunized dams, and in serum of pups born to immunized dams but not in the non-immunized group. BLG-specific IgA and IgG were again higher in high protein group and declined with time. The concentration decreased faster in the low proetein group than in the high protein groups. After immunization the pups with LBG, serum BLG-specific antibodies were not differ between rats born to immunized dams and those born to non-immunized dams. Therefore passive immunity rats received via milk as a pup had no effect on the BLG-specific antibody production later in life. This study shows the importance of protein status of mother and strongly support to the endorsement of breast feeding.

• Journal of Nutrition and Health
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• v.27 no.3
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• pp.263-271
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• 1994

The concentrations of the immunological substances in breast milk and nutritional status were studied in healthy Korean women of middle socioeconomic class. The subjects were recruited at random from obstetric clinics in Seoul. The nutrients intake, prepregnancy BMI, maternal weight gain during pregnancy were studied. The concentrations of lactoferrin(LF), lysozyme(LZ), sIgA, IgG and C3 in colostrum, in transitional milk, and in mature milk, were measured. To elucidate the effect of nutritional status on immunological substances, each components was compared on the basis of either BMI, weight gain, or protein intake. The highest concentrations of the substances were found in colostrum and decreased as lactation progressed. The decline was more prominent in IgG, C3 and sIgA, and less significant in LZ and LF. The colostrum of standard weight gain group showed higher concentrations compared to lower weight gain group. This difference became smaller as the lactation progresses. BMI and nutrient intake status had less significant effect. Lower sIgA was found in lower BMI, in lower weight gain, and lower protein intake groups compared to standard groups, which indicates sIgA is the most affected substance among the measured by nutritional status.

• BMB Reports
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• v.31 no.4
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• pp.413-417
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• 1998

Transgenic mice containing a bovine ${\beta}-Casein/Human$ lysozyme fusion gene (pBZ) were generated in order to produce human lysozyme in their milk. The expression vector was a quadripartite fusion consisting of a 2 kb upstream DNA of the bovine ${\beta}-casein$ gene, human lysozyme gene, intron II of the rabbit ${\beta}-globin$ gene, and the polyadenylation/termination signals of SV40 DNA. Fertilized mouse zygotes were microinjected with pBZ, then transferred into the oviduct of foster mothers. Out of 20 mice born, 11 survived until postweaning and three were identified as positivetransgenic by Southern blot analysis (one male and two females). The founder mice were mated to BCFl mice to produce transgenic progeny. It was confirmed by RT-PCR and Northern blot analyses that the transgene was specifically expressed in the mammary gland of the founder mice. Furthermore, the artificial introns within the transgenic RNA was proven to be correctly spliced out as judged by RT-PCR analysis. These results indicated that transgenic mice generated in this study properly expressed the human lysozyme RNA in their mammary gland.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.8
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• pp.993-999
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• 2010

Serum lysozyme gene is one of the important genes influencing the immune system as its product can cause lysis of bacterial cell wall by cleaving the peptidoglycan layer. The present investigation on the serum lysozyme gene of Indian riverine buffalo was undertaken with the objectives to identify and characterize single nucleotide polymorphic patterns by PCR-SSCP method as well as to study the effect of different genotypes on serum lysozyme activity and somatic cell count. A total of 280 animals comprising four different famous bubaline breeds (Murrah, Mehsana, Surti and Bhadawari), spread over six different farms across the country were used for this study. A 276 bp (partial intron 2, complete exon 3 and partial intron 3) fragment of lysozyme gene was screened for polymorphism using the SSCP technique. Four genotypes namely AA, AB, BC and AC were observed, out of which BC genotype was found to be the most frequent. Among these three alleles, C allele (0.38) was most prevalent in these populations. Various SSCP allelic variants were cloned for sequencing and sequences were submitted to NCBI Genbank. From the alignment of the nucleotide sequences of various allelic variants, it was found that there were differences in 12 positions among the alleles, out of which maximum variation (at 8 places) was found in the intronic region. The allele A was closer to allele-C than allele-B. Allele B was phylogenetically equidistant from both of the other alleles. Mean lysozyme activity determined in serum samples of different animals of Murrah buffalo was $27.35{\pm}2.42\;{\mu}g$ per ml of serum, whereas the mean somatic cell count was $1.25{\pm}0.13{\times}10^5$ cells per ml of milk. The SSCP pattern-wise effects of various genotypes on lysozyme activity and SCC were analyzed. Although the mean values were apparently different in various genotypes, these differences were statistically non-significant. It can be concluded that the riverine buffaloes are sufficiently polymorphic with respect to serum lysozyme gene. The absence of AA genotype in Bhadawari breed of buffalo can be considered as a marker for breed characterization. The difference of four nucleotides in exon-3 indicates high selection pressure on the gene.