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Home > Comparative Reference Values of Somatic Cells, Enzymes and Some Biochemical Constituents in the Milk from Uninfected Mammary Glands of Nili-Ravi Buffaloes, Sahiwal and Cross-Bred Cows.

Comparative Reference Values of Somatic Cells, Enzymes and Some Biochemical Constituents in the Milk from Uninfected Mammary Glands of Nili-Ravi Buffaloes, Sahiwal and Cross-Bred Cows.

Thesis Info

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Author

Kausar, Razia

Program

PhD

Institute

University of Agriculture

City

Faisalabad

Province

Punjab

Country

Pakistan

Thesis Completing Year

2015

Thesis Completion Status

Completed

Subject

Natural Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/7030/1/Razia_Kausar_Veterinary_Anatomy_UAF_2015.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725761455

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Pakistan is among the top milk producing countries of the world. However, the quality control standards for milk have not as yet been established. The present study was conducted to establish the normal reference values of milk somatic cell counts (SCC), different milk enzymes and other important milk constituents in non-infected milk of Nili-Ravi buffaloes, Sahiwal and cross-bred cows. The milk samples from 30 animals (in the first two months of first to 5th lactation) of each group were collected from Livestock Experimental Station, University of Agriculture, Faisalabad and commercial dairy farms and tested for mastitis using Surf Field Mastitis test and microbiological examination and negative samples were used to establish the reference values. Milk samples were analyzed for different parameters like pH, electrical conductivity (EC), SCC, milk enzymes i.e. lacto-peroxidase (LPO), superoxide dismutase (SOD), guaiacol peroxidase (GPO), catalase (CAT), protease, amylase, α-esterase, NAGase, total phenolic contents (TPC), proteins and malondialdehyde (MDA) content and reference values were established. Total protein (81.6±2.0 mg mL-1), casein (71.4±1.2 mg mL-1), TPC (2381.6±71.85 μM mL-1), total oxidant status (TOS) (61.25±0.59 μM mL-1), protease (81.3±3.35 U/mL), CAT (97.45±4.8 U/mL), LPO (1.75±0.06 U/mL), NAGase (56.07±2.33 U/mL) and SCC (178645.83±2324.0 mL-1) were the highest in milk of crossbred cows. Whey protein (28.8±1.25 mg mL-1), GPO (110.74±8.64 U/mL), SOD (17.15±0.56 U/mL), amylase (89.44±2.51U/mL), EC (5.7±0.04) and MDA (2.27±0.07 μM mL-1) was the highest in Sahiwal cow’s milk. Total antioxidant capacity (TAC) (3.296±0.005 mM L-1) and α-esterase (361.19±13.63 U/mL) activity were the highest in milk of Nili-Ravi buffaloes. The SCC negatively correlated with TOS, TAC, Lacto-serum protein and α-esterase activity while NAGase with lacto-serum protein and amylase, therefore, their higher values can be used as indicators of good milk quality. SCC and NAGase positively correlated with TPC, CAT and LPO. As such their lower values seem to be associated with better udder health and good milk quality. GPO and SOD negatively correlated with TOS, TAC, total protein, casein and α-esterase activity but positively correlated with amylase, EC, SCC, MDA and lacto-serum protein therefore, their lower values in milk may be desirable. Milk quality of Nili-Ravi was comparatively superior based on lower values of SCC, EC, NAGase, CAT, GPO, SOD, TPC, protease and higher TAC. Protein profiling through SDS-PAGE clearly resolved the major milk peptides. In high molecular weight (M. wt.) zone, proteins of ~208 kDa and ~190 kDa were detected in all tested samples. In medium M. wt. zone, three peptides i.e. lactoferrin (78.2kDa), serum albumin (66.2kDa) and heavy chain of immunoglobulin (IgG) (54 kDa) were detected in all samples while a prominent band of ovalbumin (45kDa) was also detected mainly in cow milk samples. In low M. wt. zone, clear bands of milk caseins were detected. All four casein (CN) bands i.e. αS2 – CN (29 kDa), αS1 – CN (27 kDa), β - CN (24 kDa) and κ- CN (22 kDa) were detected in Sahiwal and cross-bred cows. However, in milk of Nili-Ravi buffaloes, three casein protein i.e. αS2 – CN (29 kDa), β - CN (25 kDa) and κ- CN (22 kDa) were detected. In milk of Nili-Ravi buffaloes, αS1 – CN (27 kDa) was not detected. Moreover, a band of β-lactoglobulin (~18 kDa) was detected in milk of cross-bred cows and not in other samples especially those of Nili-Ravi buffaloes. As the αS1-casein and β-lactoglobulin are the major allergens, milk of Nili- Ravi buffaloes that lacks these peptides can be used for development of hypoallergenic or non-allergic dairy products. Deferential peptides may also help to differentiate the milk from different tested dairy species/cow types.
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Asthma and allergy are common and complex diseases often co-occurring within the same individual. Genetic as well as environmental factors such as consanguinity and early sensitization to allergens significantly influence the onset and progress of the disease. Aim of the following study was to investigate the association of environmental factors and selected single nucleotide polymorphisms (SNPs) from candidate genes with asthma in Pakistani population. The studied population consisted of 368 genetically unrelated volunteers. Three SNPs from 2 genes Group Specific Component gene GC (rs4588 and rs7041) and Glutathione S-Transferase P1 gene GSTP1 (rs1695) were selected for genotype analysis by Restriction fragment length polymorphism (RFLP), whereas 15 SNPs form Interleukin 33 gene IL33 (rs1412426, rs1342326, rs992969, rs928413, rs2066362, rs996029, rs10815388, rs10975501, rs17498196, rs10975516, rs10975519, rs7047921, rs1332290, rs16924241 and rs8172) were selected for ABI’s Taqman 7900 genotyping. Mean age was 34.04± 16.30 for males and 33.89±13.15 for females whereas the mean BMI was 22.01±4.04 for males and 23.23±5.21 for females. About 68.20% subjects lived in urban areas and 31.79% lived in rural areas. Parents of 87.5% of the studied population had married within their own caste system, 60.05% of which were first cousins. Family history of Asthma was prevalent in 48.91% of the Asthmatic population. Total 60.86% asthma cases were classified as adult and 39.13% cases were classified as pediatric onset asthma, based on the age at which the diagnosis of asthma was concluded. Asthma severity was based on the percent predicted FEV1 and categorized as intermittent (2.71%), mild persistent (0.54%), moderate persistent (40.21%) and severe persistent (56.52%). Total serum IgE levels were 121±2.91 lU/mI for the control population and 584.2±4.61 lU/mI in asthmatics. Males had higher IgE levels (181IU/ml) than the females (99lU/ml). Amongst the atopic manifestations cough was the most prevalent symptom (82.06%) closely followed by wheeze (80.97%) whereas skin allergies were the least prevalent (22.82%). Seasonal variations were reported to be the most significant asthma trigger (95.65%) followed by dust (72.82%) and food allergies (60.32%). Asthma attacks were more frequent in winter season (57.06%) and in transitional period between changing seasons (48.36%). Only 3.80% reported no seasonal correlations with asthma. Smoking asthmatics constituted 11.41% of the studied asthmatic population whereas 62.5% were exposed to tobacco smoke for long periods of time, 51.08% asthmatics were exposed to tobacco smoke but never smoked themselves. Homeopathic medicine was used by 14.67% and 33.69% turned to Hakeems however; the use of complimentary alternate medication remained the most popular (39.13%) mostly due to the lack of side effects associated with such treatments. Total 18 SNPs from 3 candidate genes were genotyped using RFLP and ABI’s Taqman. RFLP was carried out in 2 genes (GC/VDBP, GSTP1). Significant association of GC/VBDP GC2 homozygote asthma was established (OR = 3.14, 95% CI = 1.786 - 5.535, p = <0.001) through 90% reproducibility. GSTP1 Ile/Ile homozygote showed significant association with asthma (OR = 2.33, 95% CI = 1.347 - 4.043, p = 0.003) in Pakistani population. 15 SNPs from Interleukin-33 were genotyped on the ABI platform. rs17498196 (OR = 1.78, 95% CI = 1.18- 2.68 p = 0.005) and rs992969 (OR = 1.42, 95% CI = 0.99- 2.04 p = 0.05) were significantly associated with the risk of asthma, however the risk becomes non-significant with cousin parents removed rs17498196 (OR = 2.42, 95% CI = 1.23- 4.74 p = 0.01) and rs992969 (OR = 1.41, 95% CI = 0.80- 2.48 p = 0.23) respectively. These results highlight the importance of consanguineous unions in inheritance of asthma as a genetic disorder. LD analysis between the SNPs revealed strong linkage disequilibrium between rs10975516, rs10975519, rs7047921 and rs1332290 even with cousin parents removed remaining good predictors of each other. rs1412426, rs1342326, rs992969 and rs928413 showed moderate LD which further decreased with cousin parents removed, implicating consanguinity in the inheritance of asthma risk allele.