Molecular, biochemical and enzymatic characterization of photorespiratory 2-phosphoglycolate phosphatase (PGLP1) in rice.

Phosphoglycolate phosphatase (PGLP, EC3.1.3.18) is a key enzyme in photorespiration. However, genes encoding the rice photorespiratory PGLP have not yet been identified or characterized. Here, PGLP for photorespiration in rice was identified and its enzymatic properties were investigated. In order to define the function of PGLP homologs, rice PGLP mutants were constructed using CRISPR/Cas9, the transcriptional expressions were analyzed by RT-qPCR, and subcellular localizations were detected via rice protoplast transient expression analysis. Based on sequence alignment, proteins encoded by genes OsPGLP1, OsPGLP2, and OsPGLP3 in the rice genome were predicted to have PGLP activity. Subsequent experimentation showed that OsPGLP1 and OsPGLP3 are chloroplast proteins, while OsPGLP2 is localized in the cytoplasm. In rice leaves, levels of PGLP1 transcript were substantially higher than those of PGLP2 and PGLP3, whereas in roots, levels of PGLP2 transcript were higher than those of PGLP1 and PGLP3. There was no detectable PGLP activity in leaves of the OsPGLP1 mutant, which was non-viable in ambient air condition (400 ppm CO2 ) and high CO2 (4000 ppm) was unable to restore normal growth. In contrast, mutations of PGLP2 or PGLP3 did not result in visible phenotypes and the leaf PGLP activities were also unaffected It is suggested that PGLP1, encoded by Os04g0490800, is responsible for photorespiration. Furthermore, PGLP1 is a dimer with an apparent molecular mass of ca.65 kDa, and its Km is 272 µM, with a higher broad optimum pH (7.5 to 10.0) for PGLP activity than that in other higher plants.

Fatty acid profile of yak milk from the Qinghai-Tibetan Plateau in different seasons and for different parities.

Yaks are the most important grazing livestock for milk production on the Qinghai-Tibetan plateau because these animals are adapted to the high elevations and extremes of cold and can graze throughout the year. In the present study, 30 yaks were selected and the fatty acid (FA) profile of yak milk at different seasons and parities was investigated using gas chromatography. The concentrations of cis-9 C18:1, cis-11 C18:1, cis-9,trans-11 C18:2, and C18:3 n-3 in yak milk were higher in summer (25.26, 1.50, 1.46, and 0.33 g/100 g of total FA, respectively) than in winter (22.17, 0.77, 1.27, and 0.28 g/100 g of total FA, respectively). The contents of monounsaturated and polyunsaturated FA in milk fat of multiparous (parities 2 to 5) yaks (31.61 and 4.20 g/100 g of total FA, respectively) were higher than those in primiparous yaks (29.61 and 3.80 g/100g of total FA, respectively). These results suggest (1) that the potential exists to improve the FA composition of yak milk by developing local supplement resources during the winter and (2) that multiparous yaks have a more favorable FA profile than primiparous yaks.

Effect of somatic cell count in goat milk on yield, sensory quality, and fatty acid profile of semisoft cheese.

This study investigated the effect of somatic cell count (SCC) in goat milk on yield, free fatty acid (FFA) profile, and sensory quality of semisoft cheese. Sixty Alpine goats without evidence of clinical mastitis were assigned to 3 groups with milk SCC level of <500,000 (low), 500,000 to 1,000,000 (medium), and 1,000,000 to 1,500,000 (high) cells/mL. Thirty kilograms of goat milk with mean SCC levels of 410,000 (low), 770,000 (medium), and 1,250,000 (high) cells/mL was obtained for the manufacture of semisoft cheese for 2 consecutive weeks in 3 lactation stages. The composition of milk was analyzed and cheese yield was recorded on d 1. Cheese samples on d 1, 60, and 120 were analyzed for total sensory scores, flavor, and body and texture by a panel of 3 expert judges and were also analyzed for FFA. Results indicated that milk composition did not change when milk SCC varied from 214,000 to 1,450,000 cells/mL. Milk with higher SCC had a lower standard plate count, whereas coliform count and psychrotrophic bacteria count were not affected. However, milk components (fat, protein, lactose, casein, and total solids) among the 3 groups were similar. As a result, no significant differences in the yield of semisoft goat cheeses were detected. However, total sensory scores and body and texture scores for cheeses made from the high SCC milk were lower than those for cheeses made from the low and medium SCC milks. The difference in milk SCC levels also resulted in diverse changes in cheese texture (hardness, springiness, and so on) and FFA profiles. Individual and total FFA increased significantly during ripening, regardless the SCC levels. It is concluded that SCC in goat milk did not affect the yield of semisoft cheese but did result in inferior sensory quality of aged cheeses.

Short communication: Effects of milk fat depression induced by a dietary supplement containing trans-10, cis-12 conjugated linoleic acid on properties of semi-hard goat cheese.

Dietary supplements of conjugated linoleic acid (CLA) containing trans-10, cis-12 CLA reduce milk fat synthesis in lactating goats. This study investigated effects of milk fat depression induced by dietary CLA supplements on the properties of semi-hard goat cheese. Thirty Alpine does were randomly assigned to 1 of 3 groups and fed diets with lipid-encapsulated CLA that provided trans-10, cis-12 CLA at 0 (control), 3 (CLA-1), and 6 g/d (CLA-2). The experiment was a 3 x 3 Latin square design. Periods were 2 wk in length, each separated by 2-wk periods without CLA supplements. Bulk milk was collected on d 3 and 13 of each of 3 periods for cheese manufacture. The largest decrease (23.2%) in milk fat content, induced by the high dosage (6 g/d per doe) of trans-10, cis-12 CLA supplementation at d 13 of treatment, resulted in decreases of cheese yield and moisture of 10.2 and 10.0%, respectively. Although CLA supplementation increased the hardness, springiness, and chewiness, and decreased the cohesiveness and adhesiveness of cheeses, no obvious defects were detected and no significant differences were found in sensory scores among cheeses. In conclusion, milk fat depression induced by a dietary CLA supplement containing trans-10, cis-12 CLA resulted in changes of fat-to-protein ratio in cheese milk and consequently affected properties of semi-hard goat cheese.

Effect of extended storage on microbiological quality, somatic cell count, and composition of raw goat milk on a farm.

Dairy goat herds in the United States generally are small, widely scattered, and distant from processing facilities. Unlike the situation for cow milk, it is not cost-effective to collect goat milk everyday or every other day. In some areas, goat milk is collected only once each week, which is in violation of regulations specified in the Pasteurized Milk Ordinance for grade A milk. This study was conducted to determine the effect of up to 7 days of refrigerated bulk tank storage on composition, somatic cell count (SCC), pH, and microbiological quality of goat milk. Duplicate farm bulk tank samples were taken daily after the morning milking for seven consecutive days each month during the lactation season. Samples were analyzed immediately for all variables except free fatty acids. There were no significant changes (P > 0.05) detected in milk fat, protein, lactose, nonfat solids, SCC, or pH during extended storage, although significant effects of stage of lactation (P < 0.05) were observed. The mean standard plate count (SPC) increased to 1.8 x 10(5) CFU/ml after 6 days of storage, exceeding the grade A limit (i.e., 1.0 x 10(5) CFU/ml). The mean psychrotrophic bacteria count increased steadily to 1.5 x 10(4) CFU/ml after 6 days of storage, whereas the mean coliform count was approximately 500 CFU/ml for the first 3 days and less than 2500 CFU/ml throughout the 7 days of storage. No significant changes (P > 0.05) in the concentrations of free fatty acids, except for butyric and caprylic acids, were observed during milk storage. When stored under refrigerated and sanitary conditions, goat milk in farm bulk tanks met the grade A criteria for both SPC and SCC during 5 days of storage but was of low quality thereafter because of the growth of psychrotrophic bacteria.

Validation of antibiotic residue tests for dairy goats.

The SNAP test, LacTek test (B-L and CEF), Charm Bacillus sterothermophilus var. calidolactis disk assay (BsDA), and Charm II Tablet Beta-lactam sequential test were validated using antibiotic-fortified and -incurred goat milk following the protocol for test kit validations of the U.S. Food and Drug Administration Center for Veterinary Medicine. SNAP, Charm BsDA, and Charm II Tablet Sequential tests were sensitive and reliable in detecting antibiotic residues in goat milk. All three assays showed greater than 90% sensitivity and specificity at tolerance and detection levels. However, caution should be taken in interpreting test results at detection levels. Because of the high sensitivity of these three tests, false-violative results could be obtained in goat milk containing antibiotic residues below the tolerance level. Goat milk testing positive by these tests must be confirmed using a more sophisticated methodology, such as high-performance liquid chromatography, before the milk is condemned. LacTek B-L test did not detect several antibiotics, including penicillin G, in goat milk at tolerance levels. However, LacTek CEF was excellent in detecting ceftiofur residue in goat milk.

Proteolytic and rheological properties of aging cheddar-like caprine milk cheeses manufactured at different times during lactation.

The effects of 24 wk of aging on the proteolytic and rheological properties of cheddar-like cheese made from caprine milk collected at different lactation periods were evaluated. Cheddar cheese was made weekly using whole milk from Alpine goats and cheeses manufactured at weeks 4, 5, 12, 14, 15, 21, 22, and 23 of lactation were evaluated for proteolytic and rheological properties at 5 d after manufacture and after 8, 16, and 24 wk of aging at 4 degrees C. Rheology results indicated that a minimum of 8 wk of aging was needed to stabilize the texture of the cheese and that the most uniform cheeses were made from mid lactation milk. Cheeses manufactured at weeks 12 to 15 of lactation were the firmest, had the least flexible protein matrix (highest values for hardness, chewiness, and shear stress and rigidity at point of fracture), and the lowest degree of proteolysis. Understanding the factors that impact the texture of cheese, such as aging and the period of lactation that cheesemilk is obtained, will help develop guidance for maintaining the production of high quality and uniform caprine milk cheeses.