Fourth Flush Research Articles

QUANTITATIVE ESTIMATION OF TERPENOID CONTENT IN SOME TEA CULTIVARS OF NORTH EAST INDIA AND THEIR IN VITRO CELL CULTURES USING AN OPTIMIZED SPECTROPHOTOMETRIC METHOD

Terpenoids, which constitute one of the most diverse groups of bioactive compounds in plants, contribute to about 60% of the known natural compounds. Tea (Camellia sinensis), which is a highly preferred beverage throughout the world, is also rich in these flavor-imparting bioactive compounds. In this study, a simplified spectrophotometric method has been developed for the quantitative estimation of terpenoids in polyphenol-rich tea plant. Furthermore, terpenoid content in leaves and flowers of tea plant and their respective in vitro cultured cells have also been estimated. Terpenoid content was found to be up to five folds higher in flowers (123.9 mg/g DW) than in leaves (24.2 mg/g DW), and up to 1.7 folds higher in cultured flower cells (184.8 mg/g DW) as compared to in vitro cultured leaf cells (110.1 mg/g DW). Additionally, comparison of terpenoid content in tea leaves across the four different plucking seasons revealed a unique pattern wherein second (42.2 mg/g) and fourth flush (42.6 mg/g) tea were found to have the highest terpenoid content. First of its kind, this study provides an inexpensive method for screening of cultivars with better flavour and defense attributes. Furthermore, it also paves a way for future research in tea flowers which have high terpenoid content and the utilization of these therapeutic bioactive terpenoids in the development of diverse products for human health and wellness.

Antioxidant properties in the oyster mushrooms (Pleurotus spp.) and split gill mushroom (Schizophyllum commune) ethanolic extracts

Ethanolic extracts of oyster mushrooms (Pleurotus pulmonarius, P. ostreatus, P. djamor var. djamor and P. djamor var. roseus) and the split gill mushroom (Schizophyllum commune) were successfully investigated for their antioxidant properties. The study was carried out to determine the capability of the extracts to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals, to determine its reducing ability towards ferricyanide complex (FRAP assay) and to determine the concentration of total phenolic content (TPC) and total flavonoids content (TFC) in the macrofungi ethanolic extracts. Inhibition concentration at 50% (IC50) for each extract to scavenge DPPH radicals was detected from 2.75 mg/ml to 12 mg/ml where S. commune from the fourth flush showed the lowest IC50 value among all tested mushrooms. The greatest ability to reduce ferricyanide complex to ferrous form was observed in P. djamor var. djamor at concentration 10 mg/ml in both first (1.23 ± 0.02) and second flushes (1.23 ± 0.00). Meanwhile, the highest TPC was determined in P. djamor var. djamor extract (51.94 ± 0.04 mg TAE/g dry weight of extract) whereas S. commune gave highest reading of total flavonoid content in TFC assay (29.80 ± 0.27 mg QE/g dry weight of extract). All the mushroom samples showed appreciable antioxidant properties which therefore, can be promoted as natural antioxidant preference in food and pharmaceutical industries.

Developmental capacity of cumulus oocyte complexes collected from prepubertal cattle with and without gonadotropin stimulation employing ultrasound-guided follicular aspiration

A total of 139 laparoscopies and 106 uterine flushings were performed in 28 female sheep of three different breeds (Finnish Landrace, Merino, German Blackface). Nine 1-year-old Finnish Landrace and German Blackface sheep were used as recipients. Oestrus was synchronised by intravaginal sponges and the donors were superovulated using either 16 mg follicle stimulating hormone (FSH-P) or 1500 IU pregnant mares' serum gonadotrophin (PMSG). Embryo collections were done by laparoscopy through three stab incisions in the area anterior to the udder. A uterine horn was punctured close to the uterine body and a flexible catheter was inserted into the tip of the uterine horn. It was necessary to clamp the tip of the uterine horn with grasping forceps and flush the uterine lumen twice to obtain high recovery rates. Superovulatory treatment led to an average of 4.7 ± 4.6 corpora lutea (CL) without differences between PMSG and FSH-P. A total of 226 ovine embryos were recovered of which 144 (63.7%) were fertilised, including 65 (28.8%) damaged morulae. The remaining portion consisted of 26.5% unfertilised ova and 9.7% empty zonae pellucidae. The embryo recovery rate using laparoscopic technology was approximately 75%. A repeated embryo recovery was possible up to three times on a monthly basis with an efficiency of approximately 75–80%. The fourth flush resulted in a significantly reduced recovery rate (47.4%). The limiting factor for continued laparoscopic embryo collection was the postoperative protrusion of the endometrium at the puncture wounds of the uterine walls. Laparoscopic transfer of eight freshly collected ovine embryos to four recipients led to one pregnancy. Transfer of ten frozen/thawed embryos to five recipients led to three pregnancies. All pregnancies resulted in the delivery of normal lambs.

Nitrogen Nutrition of Containerized Citrus Nursery Plants

Experiments were conducted with `Hamlin' orange [Citrus sinensis (L.) Osb.] budded on Cleopatra mandarin (Citrus reticulata Blanco) or Carrizo citrange [Citrus sinensis (L.) Osb. × Poncirus trifoliata (L.) Raf.] seedling rootstocks to determine minimum container solution N concentrations required for optimum growth and fertilizer uptake efficiency at various growth stages. Plants were fertigated daily with 1 liter of N solution at either 0, 12.5, 25, 50, 100, or 200 mg·liter-1 from NH4NO3 or 0, 3.13, 6.25, 12.5, 25, or 50 mg·liter-1 from NH4NO3 dissolved in a complete nutrient solution, respectively. Percentage of N in the mature plant tissues increased as N concentration in the medium solution increased. Shoot length and leaf area increased as N concentrations increased up to a critical concentration of 15 to 19 mg·liter-1. The critical N concentration for root, shoot, and total plant dry weight was ≈18 mg·liter-1 for `Hamlin'-Cleopatra mandarin nursery plants and 15 mg·liter-1 for `Hamlin'-Carrizo nursery plants. The critical N concentration for relative total plant dry weight accumulation (percentage) for the two experiments was 16.8 mg·liter-1. In a separate experiment, plants were given labeled fertilizer N (FN) (15NH415NO3) at one of five growth stages: A) in the middle of rapid shoot extension of the third flush, B) immediately following the cessation of the third flush shoot extension but during leaf expansion, C) immediately following leaf expansion, D) before the fourth flush, or E) in the middle of rapid shoot extension of the fourth flush. Labeled FN recovery increased during rapid shoot extension of the fourth scion flush compared to the other labeling periods. FN uptake per gram of total plant dry weight was greatest during rapid shoot extension (A and E) and lowest during the intermediate labeling periods (B-D). FN supplied 21% to 22% of the N required for new growth during rapid shoot extension.

Collection and transfer of ovine embryos by laparoscopy

A total of 139 laparoscopies and 106 uterine flushings were performed in 28 female sheep of three different breeds (Finnish Landrace, Merino, German Blackface). Nine 1-year-old Finnish Landrace and German Blackface sheep were used as recipients. Oestrus was synchronised by intravaginal sponges and the donors were superovulated using either 16 mg follicle stimulating hormone (FSH-P) or 1500 IU pregnant mares' serum gonadotrophin (PMSG). Embryo collections were done by laparoscopy through three stab incisions in the area anterior to the udder. A uterine horn was punctured close to the uterine body and a flexible catheter was inserted into the tip of the uterine horn. It was necessary to clamp the tip of the uterine horn with grasping forceps and flush the uterine lumen twice to obtain high recovery rates. Superovulatory treatment led to an average of 4.7 ± 4.6 corpora lutea (CL) without differences between PMSG and FSH-P. A total of 226 ovine embryos were recovered of which 144 (63.7%) were fertilised, including 65 (28.8%) damaged morulae. The remaining portion consisted of 26.5% unfertilised ova and 9.7% empty zonae pellucidae. The embryo recovery rate using laparoscopic technology was approximately 75%. A repeated embryo recovery was possible up to three times on a monthly basis with an efficiency of approximately 75–80%. The fourth flush resulted in a significantly reduced recovery rate (47.4%). The limiting factor for continued laparoscopic embryo collection was the postoperative protrusion of the endometrium at the puncture wounds of the uterine walls. Laparoscopic transfer of eight freshly collected ovine embryos to four recipients led to one pregnancy. Transfer of ten frozen/thawed embryos to five recipients led to three pregnancies. All pregnancies resulted in the delivery of normal lambs.

Effects of the paedogenetic mushroom cecid, Heteropeza pygmaea (Diptera:Cecidomyiidae) on cropping of the cultivated mushroom (Agaricus bisporus)

SUMMARYWhen introduced into a mushroom crop at rates of 2, 20 or 200 larvae/tray (0.56 m2), the mushroom cecid, Heteropeza pygmaea, caused significant reductions in both yield and number of mushrooms in relation to the infestation level. The reductions were greater when the larvae were introduced at spawning rather than at casing. The yield and number of infested (unmarketable) mushrooms increased significantly in relation to the initial infestation level.Just two H. pygmaea larvae, introduced at spawning, resulted in cecid populations that caused a 12% loss in total yield in addition to a 7% loss due to spoilage. Loss assessment in the future, therefore, should take into account both yield suppression and spoilage.There was little effect of cecid infestation on flush timing and mushroom size was only affected in the fourth flush, when a significant reduction (27%) was shown at the highest infestation rate at spawning.

Influence of a dominant follicle atresia and estradiol release by ovarian follicles during superovulation in cattle

A total of 139 laparoscopies and 106 uterine flushings were performed in 28 female sheep of three different breeds (Finnish Landrace, Merino, German Blackface). Nine 1-year-old Finnish Landrace and German Blackface sheep were used as recipients. Oestrus was synchronised by intravaginal sponges and the donors were superovulated using either 16 mg follicle stimulating hormone (FSH-P) or 1500 IU pregnant mares' serum gonadotrophin (PMSG). Embryo collections were done by laparoscopy through three stab incisions in the area anterior to the udder. A uterine horn was punctured close to the uterine body and a flexible catheter was inserted into the tip of the uterine horn. It was necessary to clamp the tip of the uterine horn with grasping forceps and flush the uterine lumen twice to obtain high recovery rates. Superovulatory treatment led to an average of 4.7 ± 4.6 corpora lutea (CL) without differences between PMSG and FSH-P. A total of 226 ovine embryos were recovered of which 144 (63.7%) were fertilised, including 65 (28.8%) damaged morulae. The remaining portion consisted of 26.5% unfertilised ova and 9.7% empty zonae pellucidae. The embryo recovery rate using laparoscopic technology was approximately 75%. A repeated embryo recovery was possible up to three times on a monthly basis with an efficiency of approximately 75–80%. The fourth flush resulted in a significantly reduced recovery rate (47.4%). The limiting factor for continued laparoscopic embryo collection was the postoperative protrusion of the endometrium at the puncture wounds of the uterine walls. Laparoscopic transfer of eight freshly collected ovine embryos to four recipients led to one pregnancy. Transfer of ten frozen/thawed embryos to five recipients led to three pregnancies. All pregnancies resulted in the delivery of normal lambs.

Effects of repeated superovulation and flushing on reproductive performance of Bos indicus cows

To determine the effect of repeated superovulation and flushing in Bos indicus cows, 1,841 embryo collection records of 813 Brahman donor cows were analyzed using least-squares methods. Number of flushes affected total number of ova collected per donor (P < 0.07); number of transferable embryos (P < 0.001); pregnancy rate in the recipient (P < 0.03); number of embryos collected per donor (P < 0.003); number of morulas (P < 0.10); and number of blastocysts (P < 0.10). Total ova recovered decreased from 9.4 at the first flush to 2.9 at the fifth flush. Least-squares means (LSM) of transferable embryos per flush ranged from 5.3 for the first flush to 2.2 on the fifth flush. Pregnancy rate in the recipient was higher (44.2%) during the fourth flush and then decreased (38.8%) after six or more flushes. LSM of number of embryos collected ranged from 6.8 during the first flush to 3.0 for the fifth flush. Embryo production was (P < 0.003) higher (6.5) during the first flush than during the second flush (5.0). Number of blastocysts recovered per donor was higher (3.9) during the first flush and lower (2.2) during the fifth flush. Repeatabilities of the ovarian response to superovulation ranged from 0.10 to 0.17. These low levels of repeatability do not encourage any culling program based strictly upon ovarian response to superovulation. These data establsih that repeated superovulation and flushing have a detrimental effect on embryo parameters, especially after the fourth flush, in Brahman donor cows.

Studies on genotoxic effects of superovulation in preimplantation mouse embryos using the sister chromatid exchange (SCE) assay: [formula omitted] Bundesgesundheitsamt, D-1000 Berlin 33

A total of 139 laparoscopies and 106 uterine flushings were performed in 28 female sheep of three different breeds (Finnish Landrace, Merino, German Blackface). Nine 1-year-old Finnish Landrace and German Blackface sheep were used as recipients. Oestrus was synchronised by intravaginal sponges and the donors were superovulated using either 16 mg follicle stimulating hormone (FSH-P) or 1500 IU pregnant mares' serum gonadotrophin (PMSG). Embryo collections were done by laparoscopy through three stab incisions in the area anterior to the udder. A uterine horn was punctured close to the uterine body and a flexible catheter was inserted into the tip of the uterine horn. It was necessary to clamp the tip of the uterine horn with grasping forceps and flush the uterine lumen twice to obtain high recovery rates. Superovulatory treatment led to an average of 4.7 ± 4.6 corpora lutea (CL) without differences between PMSG and FSH-P. A total of 226 ovine embryos were recovered of which 144 (63.7%) were fertilised, including 65 (28.8%) damaged morulae. The remaining portion consisted of 26.5% unfertilised ova and 9.7% empty zonae pellucidae. The embryo recovery rate using laparoscopic technology was approximately 75%. A repeated embryo recovery was possible up to three times on a monthly basis with an efficiency of approximately 75–80%. The fourth flush resulted in a significantly reduced recovery rate (47.4%). The limiting factor for continued laparoscopic embryo collection was the postoperative protrusion of the endometrium at the puncture wounds of the uterine walls. Laparoscopic transfer of eight freshly collected ovine embryos to four recipients led to one pregnancy. Transfer of ten frozen/thawed embryos to five recipients led to three pregnancies. All pregnancies resulted in the delivery of normal lambs.

The effect of sciarid larvae (Lycoriella auripila) on cropping of the cultivated mushroom (Agaricus bisporus)

SUMMARYSignificant linear relationships between the mean number of sciarid, Lycoriella auripila, larvae/125 g‐sample of casing and yield, numbers of mushrooms and weight/mushroom were demonstrated at all stages of a mushroom crop. Negative relationships were obtained for yield throughout the cropping period. There was no injury threshold for this pest, although an economic threshold of one larva/sample was deduced. Loss in yield was mostly due to the destruction of mushroom primordia and presumed interruption of nutrient supply to the developing sporophores.Numbers of mushrooms were severely reduced in the first, second and third flushes, more so than yield, although a large increase in numbers was demonstrated in the fourth flush. The size of mushroom was inversely related to numbers, although the increases in size in the first three flushes were insufficient to compensate for the reduction in numbers. A reduction in size was evident in the fourth flush.

Effects of bendiocarb and diflubenzuron on mushroom cropping

SUMMARYWhen mixed into the casing or compost layers of a mushroom bed in the absence of pests, bendiocarb decreased yield and number of mushrooms according to concentration. The most severe effects were on mushroom number at the two highest rates used (100 and 1000 μg/g), and there were large increases in mushroom size. Effects of bendiocarb incorporation in the compost diminished with time, and there was partial compensation in yield and numbers at the fourth flush. The action of bendiocarb persisted when it was mixed into the casing.Diflubenzuron showed some opposite effects at lower concentrations. When either mixed into, or drenched onto the casing at the commercial rate (30 μg/g), yield and size were both increased and the timing of the flushes was unaffected. At the two higher concentrations (180 and 1080 μg/g), reductions in yield and number and an increase in mushroom size were shown. However, these effects became more severe with time, especially those on mushroom number, possibly due to the accumulation of a toxic breakdown product.

Variation of psilocybin and psilocin levels with repeated flushes (harvests) of mature sporocarps of Psilocybe cubensis (earle) singer

Analysis of Psilocybe cubensis (Earle) Singer grown in controlled culture showed that the level of psilocin was generally zero in the first (or sometimes even the second) fruiting of the mushroom from a given culture and that the level reached a maximum by the fourth flush. The level of psilocybin, which was nearly always at least twice the level of psilocin, showed no upward or downward trend as fruiting progressed, but was variable over a factor of four. Samples obtained from outside sources had psilocybin levels varying by over a factor of ten from one collection to the next.

The effect of hedging time on regrowth and flowering of mature Valencia orange trees

Thirty-eight-year-old Valencia orange trees on Rough Lemon rootstock were hedged at monthly intervals from September 1973 to April 1974. Trees were hedged on the east and west sides. Size of cut stem, number and length of each flush, and fruit number were recorded for each flush. Regrowth rate was highest on trees hedged in mid-summer, but by the winter of 1976 first flush length was similar on all treatments, and there were no significant differences in the total length per shoot or in the number of first or second flush shoots. The number of third and fourth flush shoots were reduced on autumn hedged trees. Stem circumference, rather than hedging time, was the major determinant of regrowth vigour. Approximately 50% of stems cut before December had flowering regrowth next spring. In the following year all hedging treatments produced flowering regrowth. It was concluded that mature Valencia citrus should be hedged after harvest and between spring and mid-summer. Autumn hedging should be avoided because it results in reduced fruiting.

Principles of insecticide action on mushroom cropping: incorporation into casing

SUMMARYThe incorporation of organophosphorous insecticides into the casing layer of mushroom beds reduces the numbers of mushrooms, according to a logistic relationship with log10 (dose). The cumulative numbers at any flush are affected in this way, and the response of individual flushes may, therefore, be represented by the difference between two logistic curves. This explains the observed increases in numbers in the second and third flushes following diazinon treatment and in the fourth following thionazin, pirimiphos‐methyl and pirimiphos‐ethyl. No compensation in numbers followed chlorfenvinphos treatment. This decreasing response pattern reflected the increasing residual action of the insecticides.Mushroom size was inversely related to numbers but even when numbers in any flush were unaffected by the treatment, there was usually an increase in size. Thus yield increases occurred in most flushes at commercial concentrations of four insecticides (thionazin compensated only in the fourth flush) and total yields were increased by 8 to 13%.