The relationship between fetal and maternal selenium concentrations in sheep and goats

In this study, biological samples (slaughterhouse material) were collected from 30 sheep and 36 goats and classified according to gestational stage into either early or late gestation. Samples consisted of allantoic fluid, amniotic fluid, fetal liver, fetal kidney, fetal thyroid gland, maternal plasma and liver to determine selenium (Se) concentrations throughout gestation. The Se concentrations in the allantoic fluid, fetal liver and kidney increased significantly (p < 0.01) during late gestation. Concurrently, the Se concentrations in amniotic fluid, maternal plasma and liver decreased significantly (p < 0.01) over time. Significant (p < 0.01) positive relationships were recorded between the age of the fetus and Se concentrations in the allantoic fluid (r = 0.57–0.75), fetal liver (r = 0.43–0.59) and kidney (r = 0.80–0.81) in both sheep and goats. A significant (p < 0.05) positive relationships were also recorded between the Se concentrations in the allantoic fluid and fetal liver (r = 0.35–0.37), the maternal plasma and liver Se concentrations (r = 0.37–0.57) between sheep and goats. A significant (p < 0.05) negative correlation was recorded between the Se concentrations in the allantoic fluid with maternal plasma of sheep (r = −0.41) as well as between the fetal liver and maternal liver Se (r = −0.22 to 0.50) and a negative correlation (r = −0.42 to 0.43) (p < 0.01) between Se concentrations in the fetal liver and amniotic fluid in both sheep and goats, respectively. Se concentration in the fetal liver was significantly (p < 0.01) higher than that of the kidney and thyroid. In the thyroid gland no morphological differences were noted. Strong fetal–maternal relationships in Se concentration were evident throughout the gestational period and dams seem to sacrifice Se levels in order to maintain that in the fetus. Se concentrations in the amniotic and allantoic fluids could be used as a possible indicator of the Se status of the fetus throughout gestation.     

Domoic acid exposure and associated clinical signs and histopathology in Pacific harbor seals (Phoca vitulina richardii)

Domoic acid (DA) is a potent neurotoxin that has caused strandings and mortality of seabirds and marine mammals off the California coast. Pacific harbor seals (Phoca vitulina richardii) are an abundant, nearshore species in California; however, DA exposure and toxicosis have not been documented for harbor seals in this region. To investigate DA exposure in harbor seals, samples were collected from free-ranging and stranded seals off California to assess exposure, clinical signs of toxicosis, and brain lesions in harbor seals exposed to DA. Domoic acid was detected in 65% (17/26) of urine samples collected from apparently healthy free-ranging seals, with concentrations of 0.4–11.7 ng/ml. Domoic acid also was detected in feces (2.4–2887 ng/g), stomach contents (1.4 ng/g; stranded only), milk (2.2 ng/ml; stranded only), amniotic fluid (9.7 ng/ml; free-ranging only), fetal meconium (14.6–39.8 ng/g), and fetal urine (2.0–10.2 ng/ml). Clinical signs indicative of DA toxicosis were observed in two live-stranded seals, and included disorientation, seizures, and uncoordinated movements. Histopathology revealed the presence of brain lesions consistent with DA toxicosis in two live-stranded seals, and one free-ranging seal that died during capture. Results indicated that harbor seals were exposed to DA, exhibited clinical signs and histological lesions associated with DA exposure, and that pups were exposed to DA in utero and during lactation via milk. Future investigation is required to determine the magnitude of impact that DA has on the health and mortality of harbor seals.     

Cryopreservation of rainbow trout Oncorhynchus mykiss spermatozoa: Effects of extender supplemented with different antioxidants on sperm motility,velocity and fertility

In present study, it was examined whether addition of different antioxidants to the cryopreservation extenders had an effect on semen post-thaw fertility and motility in rainbow trout (Oncorhynchus mykiss) and also it was investigated the sperm characteristics post-thaw sperm characteristics and fertility. The collected semen was pooled to minimize individual variation. Each pooled ejaculate was split into 12 equal aliquots and diluted with base extenders supplemented with the antioxidants, and a base extender with no additives (control). The pooled semen samples diluted at the ratio of 1:10 by the extenders were subjected to cryopreservation. Antioxidants were separately added to the extenders (one per experimental group): catalase (250 U/l), superoxide dismutase (250 U/l), peroxidase (250 U/l), oxidized glutathione (1.5 mmol/l), reduced glutathione (1.5 mmol/l), l-methionine (1.5 mmol/l), uric acid (0.25 mmol/l), l-ascorbic acid (0.5 mmol/l), α-tocopherol (2.0 mmol/l), β-carotene (0.5 mmol/l) and carnitine (0.5 mmol/l). After dilution the semen was aspirated into 0.25 ml straws, the straws were placed on the tray, frozen for 10 min, and plunged into liquid nitrogen. Our results indicated that the post-thaw motility rate increased in extenders supplemented with uric acid, l-methionine, SOD, l-carnitine, α-tocopherol and l-reduced glutathione (p < 0.05). The motility duration of frozen thawed semen increased in extenders supplemented with uric acid, l-methionine, SOD, α-tocopherol and l-reduced glutathione (p < 0.05). Fertilization rate and hatching rate of frozen-thawed semen was not affected by the tested antioxidants. Consequently, the tested antioxidants affected the motility parameters and cryopreservation extenders could be supplement with antioxidants. This study suggested usage of antioxidants in the cryopreservation of rainbow trout.     

Effects of dose and adaptation time of a specific blend of essential oil compounds on rumen fermentation

Essential oils can be used as natural additives in animal feeds. The present study evaluated the effects of three different doses and different adaptation times of a specific blend of essential oils (BEO) on rumen microbial fermentation. Eight dual flow continuous culture fermenters (1320 ml) were used in two periods of 9 days each to study the effects of increasing doses of BEO. Treatments were: CTR (no BEO), D5 (5 mg/l of BEO), D50 (50 mg/l of BEO) and D500 (500 mg/l of BEO). During the last 3 days, samples were taken at 0, 2, 4 and 6 h after the morning feeding and analyzed for large peptide (LPep), small peptides plus amino acid (SPep + AA) and ammonia N concentrations, and at 2 h after feeding for volatile fatty acids (VFA) concentration and profile. The D5 increased total VFA concentration, acetate proportion and acetate to propionate ratio, and decreased propionate and valerate proportion, compared with CTR. The concentration of LPep N tended (P=0.08) to be lower for D5 compared with CTR. In the second experiment, eight sheep were used to study the effects of long-term adaptation of rumen fluid to BEO on ruminal fermentation. Four sheep were assigned at random to the CTR treatment (no BEO) and four sheep were adapted to BEO (110 mg/day of BEO) for 4 weeks (ADBEO). After 4 weeks samples of ruminal fluid were obtained at 0 and 3 h after the morning feeding and in 2 consecutive days using an oro-ruminal probe. Samples were analyzed for LPep, SPep + AA and ammonia N concentrations, total and individual VFA, and pH. Treatment ADBEO tended (P<0.10) to increase acetate proportion and decrease valerate proportion, compared with CTR. Ruminal fluid collected from each of CTR and ADBEO sheep was used to study in vitro fermentation profile of soybean meal, corn meal, alfalfa hay and ryegrass hay. Treatments were: Control fluid (CTR without BEO), CTR fluid plus a single dose of BEO (11 mg/l; CTR + BEO) and ADBEO fluid plus a single dose of BEO (11 mg/l; ADBEO + BEO). Acetate proportion and acetate to propionate ratio was higher, and propionate and isovalerate proportion, and BCVFA and ammonia N concentration were lower in ADBEO + BEO fluid compared with CTR fluid. The addition of essential oils can shift the microbial fermentation in the rumen by increasing the acetate to propionate ratio and inhibiting deamination.     

Blood lead level and its relationship to essential elements in preschool children from Nanning,China

ObjectiveOur study aimed to assess the distribution of blood lead level and its relationship to essential elements in preschool children in an urban area of China.Design and methodsA total of 6741 children aged 0- to 6-year-old were recruited. Levels of lead, zinc, copper, iron, calcium, and magnesium in whole blood samples were determined using atomic absorption spectrometry.ResultsThe mean blood lead level (BLL) and the prevalence of BLL ≥ 10 μg/dl (5.26 ± 4.08 μg/dl and 6.84%, respectively) increased with age gradually, and there was a gender-difference for blood lead, copper, zinc and iron levels. Compared with the group of children who had BLLs < 5 μg/dl, the groups of 5 ≤ BLLs < 10 μg/dl and 10 ≤ BLLs < 15 μg/dl showed higher blood zinc, iron and magnesium levels, and a lower blood calcium level. A positive correlation of lead with zinc, iron and magnesium, and a negative correlation of lead with calcium were found in the group of children with BLL < 5 μg/dl.ConclusionAge- and gender-differences were found when assessing the BLL and intoxication prevalence in preschool children. Metabolic disorder of essential elements was found even with a low level of lead exposure.     

Copper,zinc and iron levels in premature infants following red blood cell transfusion

This study aimed to investigate effect of erythrocyte suspension (ES) transfusion on Cu, Zn, and Fe levels. It was conducted on 53 premature infants who were admitted to Hacettepe Hospital and received EST for first time. Blood samples were drawn before and 96 h after ES transfusion to determine Cu, Zn, and Fe levels in plasma and/or erythrocytes. The mean plasma Cu levels were 99 ± 3 μg/dl and 113 ± 3 μg/dl; Zn levels were 105 ± 2 μg/dl and 115 ± 23 μg/dl; mean plasma Fe level was 58.1 ± 19.4 and 75.2 ± 25.4 μg/dl and mean erythrocyte Fe level was 4182 ± 2314 μg/ml and 7009 ± 5228 μg/ml, before and after ES transfusion. The differences between before and after ES transfusion in Cu, Zn and Fe levels were significant. Correlation between plasma and erythrocyte Fe levels was significant both before and after ES transfusion. Though Fe overload is a major cause of morbidity/mortality after ES transfusion, alterations in trace elements should also be considered when transfusing blood to infants and children.     

Effect of the supplementation of a high-concentrate diet with sunflower and fish oils on ruminal fermentation in sheep

This study was conducted to test the hypothesis that the supplementation of a high-concentrate diet with lipids, reportedly a good strategy for improving the nutritional value of ruminant-derived products, may not necessarily be associated with detrimental effects on ruminal fermentation in sheep. Four ruminally cannulated adult ewes were fed a high-concentrate diet, with no oil (Control diet), for a 14-day adaptation period. Afterwards, they were fed the same basal diet but supplemented with sunflower oil [20 g/kg fresh matter (FM)] and fish oil (10 g/kg FM) (SOFO diet) for a further 11 days, to investigate the impact of the addition of oils on the ruminal fermentation of the diet. On days 0 (Control), 3 and 10 of the experimental period rumen fluid was sampled at 0, 1.5, 3, 6 and 9 h after the morning feeding, for analysis of pH, and ammonia, lactate and total volatile fatty acid (VFA) concentrations. Alfalfa hay was incubated in situ, using the nylon bag technique, for 12 and 24 h to examine the effect of oil supplementation on ruminal disappearance of dry matter (DM), crude protein (CP) and neutral-detergent fibre (NDF). On days 0 and 11, rumen fluid was collected just before the morning feeding and used to incubate alfalfa hay and the Control and SOFO diets by means of the in vitro gas production technique. The mean concentrations of acetate (87.8 mmol/L vs. 73.7 mmol/L) and butyrate (21.2 mmol/L vs. 17.7 mmol/L) were reduced by oil supplementation (P < 0.05) and the total VFA showed a tendency (P = 0.098) to be lower with the SOFO diet (139.0 mmol/L vs. 122.1 mmol/L). However, none of the other in vivo ruminal fermentation parameters were affected by the treatment (P > 0.10). The oil supplementation affected neither in situ rumen disappearance of DM, CP and NDF of alfalfa hay, nor rates of gas production (P > 0.10). On the other hand, a little, but significant reduction in cumulative gas production was observed when the experimental diets were incubated with rumen fluid derived from animals fed the oil-rich diet (P < 0.05).Overall, the results suggest that the supplementation of high-concentrate diets with sunflower oil (20 g/kg FM) plus fish oil (10 g/kg FM) had little effect on ruminal fermentation and therefore its use to improve the nutritional value of ruminant-derived products cannot be precluded.     

The adipokine preadipocyte factor-1 is downregulated in preeclampsia and expressed in placenta

BackgroundAdipokines contribute to the development of preeclampsia (PE), a severe pregnancy complication which increases the future risk for cardiovascular and metabolic disease in both mother and newborn. Pre-adipocyte factor-1 (Pref-1) was recently introduced as a novel antiangiogenic and antiadipogenic adipokine.Material and methodsPref-1 was quantified in patients with PE (n = 51) and healthy pregnant controls (n = 51) during pregnancy, as well as 6 months after delivery (study population 1). Furthermore, Pref-1 was investigated in the immediate peripartal period and the placenta in 40 healthy pregnant women undergoing elective cesarean section (study population 2).ResultsIn study population 1, median Pref-1 serum concentrations during pregnancy were significantly lower in women with PE (0.5 μg/l) as compared to healthy pregnant controls (0.7 μg/l) (p < 0.001). Furthermore, Pref-1 serum concentrations were independently predicted by PE, leptin levels, and gestational age in this population. In both study populations, Pref-1 serum levels significantly decreased after delivery as compared to prepartal levels. Moreover, significant expression of Pref-1 was detected in placental tissue.ConclusionMaternal Pref-1 serum concentrations are significantly decreased in PE. The pathophysiological significance of this regulation needs to be studied in more detail in future experiments.     

Plants and testate amoebae as environmental indicators in cupriferous peatlands,New Brunswick,Canada

Vegetation, testate amoebae, and metal concentrations in water and soil (mostly peat) were studied in two copper-rich treed swamps located north of Sackville, New Brunswick, Canada. One of the sites is partly disturbed, characterized by bare soil nearly devoid of vegetation cover except for isolated patches of the moss species, Pohlia nutans, around seepages and small streams. Copper concentrations in soil and water samples were high but varied among plots. Values in soil samples were as high as 16,000 μg/g in the open area, with 4550 μg/g being the mean. The highest value in groundwater was 1540 μg/l, with 292 μg/l being the mean. Twenty-seven testate amoebae species were identified from soil samples. The most abundant species were Cyclopyxis arcelloides and Centropyxis spp. Principal component analysis and detrended correspondence analysis showed that their abundance was especially high in the open area where copper concentrations were high, while species diversity of testate amoebae was low in the open area. This study suggests potential use of mosses and testate amoebae as bio-indicators and bio-monitoring tools for metals such as copper.     

The effect of glucose and maltose concentrations on pyruvate and ascorbate production,antioxidant enzyme activities and LPO levels in Fusarium equiseti

Changes in pyruvate and ascorbate production and antioxidant enzyme activities together with lipid peroxidation levels in Fusarium equiseti were investigated in relation to changes in the concentrations of glucose and maltose as carbon sources in the range of 5–25 g/l in Armstrong Fusarium Medium (AFM). The highest pyruvate concentration obtained at 20 g/l maltose was 67.5 ± 0.69 μg/ml while ascorbic acid reached a maximum value at 25 g/l glucose of 1866±26.1 μg/ml The maximum superoxide dismutas (SOD) activities related to increased pyruvate production were determined in AFM medium containing 20 g/l glucose as 41.49±0.65 and maltose as 61.12±0.8 IU/mg. Catalase (CAT) activity variations showed coherence with SOD activity in a medium containing maltose and reached 219.11±2.8 IU/mg while they were decreased with increasing glucose concentration. glutathione peroxidase (GSH-Px) activities in F. equiseti did not change significantly with glucose and maltose concentration and were determined to be 1.21±0.22 and 1.67±0.15 IU/mg, respectively. Minimum lipid peroxidation levels for each carbon source were determined in both 20 g/l maltose and glucose concentrations as 0.9 and 1.62 nmol MDA/g wet weight.     

Hiperparatiroidismo secundario en el cáncer de próstata avanzado

Background and ObjectiveHigh parathyroid hormone (PTH) concentrations are associated with increased bone resorption and bone matrix degradation. Some studies show elevated PTH concentrations and hypocalcemia in patients with advanced prostate carcinoma, although the pathophysiological significance of these findings is not well defined.Materials and methodsWe performed a retrospective study of 60 patients diagnosed with advanced prostate cancer (44 nonmetastatic and 16 metastatic) treated with androgen deprivation. In all patients, PTH, calcium, phosphorus, 25 (OH) vitamin D and prostate-specific antigen (PSA) were determined. Bone scintigraphy had previously been performed.ResultsIn patients with bone metastases, mean concentrations were as follows: calcium 9.19 mg/dl, phosphorus 3.47 mg/dl, 25 (OH) vitamin D 13.85 ng/ml, PTH 66.8 pg/ml and total PSA 101.27 ng/ml. For those without bone metastases, the results were calcium 9.39 mg/dl, phosphorus 3.38 mg/dl, 25 (OH) vitamin D 20.50 ng/ml, PTH 52.23 pg/ml and total PSA 2.52 ng/ml. PTH levels were significantly higher in patients with prostate cancer and bone metastases than in those without metastases (p=0.03). Vitamin D levels were also significantly lower in this group (p=0.03). There were no differences in other values.ConclusionsThe present study found increased PTH concentrations in patients with advanced prostate cancer. This finding could be useful to predict disease progression.     

Pregnancy diagnosis based on the fecal progesterone concentration in beef and dairy heifers and beef cows

The present study was undertaken to examine whether pregnancy diagnosis was possible by measuring fecal progesterone concentrations in beef and dairy heifers and beef cows. Rectal fecal samples collected on days 18–24 after insemination or days 11–17 after embryo transfer were mixed with methanol and shaken for preparation of a fecal solution. After centrifugation, the supernatant was extracted with petroleum ether followed by an enzyme immunoassay for progesterone. All pregnant animals showed fecal progesterone concentrations greater than 50 ng/g of fecal material on days 18–24 after AI or estrus. In non-pregnant animals, however, the fecal progesterone concentrations ranged widely from 5 to 180 ng/g of fecal material. In non-pregnant cattle, the percentage of cattle with <50 ng progesterone/g of fecal material compared with the total number was 37–60% on days 18–20, whereas the percentages increased more than 70% to a maximum of 78.1% on day 23. When 50 ng/g was considered as the cut-off value, the sensitivity and specificity of positive pregnancy tests were less than 70% on days 21–24, and 100% for negative pregnancy tests on days 18–24. There were significant differences in the mean fecal progesterone concentrations between pregnant and non-pregnant cattle on days 19–24. These results suggest that feces can be utilized to substitute for plasma and milk to measure progesterone for the purpose of pregnancy diagnosis in heifers and cows.     

Biochemical changes in the follicular fluid of the dominant follicle of high producing dairy cows exposed to heat stress early post-partum

High yielding dairy cows experience a negative energy balance (NEB) early post-partum and it was hypothesized that this may be aggravated under summer heat stress (HS) conditions. In this study, which was performed in Egypt, 20 Holstein cows were followed during summer (n = 10) and winter (n = 10) seasons. All cows were multiparous and kept at the same herd. Blood was sampled from each cow starting 1 week before the expected calving date and then at 1-week intervals until week 6 post-partum. From week 2 to 6 post-partum follicular fluid was collected through transvaginal follicular fluid aspiration at 6 days intervals. Ambient air temperature (AT) and relative humidity (RH) were recorded and temperature–humidity index (THI) was calculated as well. Respiration rate (RR), rectal temperature (RT), and body condition score (BCS) were recorded for each cow at the time of blood sampling. Concentrations of glucose, insulin like growth factor-1 (IGF-1), non-esterified fatty acids (NEFA), urea and total cholesterol (TC) were measured in each blood and follicular fluid sample. All the cows showed a significantly higher RR and RT in summer (95.5 ± 1.1 and 39.88 ± 0.06, respectively) than in winter (43.89 ± 0.61 and 38.94 ± 0.07, respectively) (P < 0.001). Body condition score loss during the early post-partum period was higher in summer than in winter (1.1 ± 0.07 vs. 0.85 ± 0.06 point, respectively) (P < 0.001). The average dominant follicle diameter was significantly lower in summer than in winter during the period of negative energy balance (11.6 ± 0.7 mm vs. 15.3 ± 1.2 mm, respectively) (P < 0.01). Under summer heat stress, the concentrations of glucose (2.98 ± 0.07 and 2.19 ± 0.04 mmol/L), IGF-1 (106.7 ± 2.9 and 99.0 ± 3.4 ng/ml) and TC (137.3 ± 5.3 and 62.2 ± 5.1 mg/dl) in blood and FF, respectively, were significantly lower than winter concentrations by (0.17 ± 0.03 mmol/L, P < 0.001 and 0.26 ± 0.06 mmol/L, P < 0.001), (12.3 ± 3.6 ng/ml, P < 0.001 and 9.0 ± 2.7 ng/ml, P < 0.001) and (20.7 ± 1.8 mg/dl, P < 0.001 and 7.3 ± 1.1 mg/dl, P < 0.01), respectively. However, the concentrations of NEFA (0.68 ± 0.14 and 0.22 ± 0.02 mmol/L) and urea (9.27 ± 0.34 and 9.96 ± 0.25 mmol/L) in blood and FF, respectively, were significantly higher in summer compared to winter (0.50 ± 0.08 mmol/L, P < 0.001 and 0.20 ± 0.02 mmol/L, P < 0.001) and (8.77 ± 0.23 mmol/L, P < 0.05 and 8.96 ± 0.29 mmol/L, P < 0.001), respectively, throughout the experimental period. The results of the present study indicate that heat stress early post-partum aggravates NEB in high yielding dairy cows, reduces BCS, dominant follicle diameter and alters the biochemical concentrations in the follicular fluid of the dominant follicle which may result in inferior oocyte and granulosa cell quality and hence poorer fertility.     

The neonicotinoids thiacloprid,imidacloprid, and clothianidin affect the immunocompetence of honey bees (Apis mellifera L.)

A strong immune defense is vital for honey bee health and colony survival. This defense can be weakened by environmental factors that may render honey bees more vulnerable to parasites and pathogens. Honey bees are frequently exposed to neonicotinoid pesticides, which are being discussed as one of the stress factors that may lead to colony failure. We investigated the sublethal effects of the neonicotinoids thiacloprid, imidacloprid, and clothianidin on individual immunity, by studying three major aspects of immunocompetence in worker bees: total hemocyte number, encapsulation response, and antimicrobial activity of the hemolymph. In laboratory experiments, we found a strong impact of all three neonicotinoids. Thiacloprid (24 h oral exposure, 200 μg/l or 2000 μg/l) and imidacloprid (1 μg/l or 10 μg/l) reduced hemocyte density, encapsulation response, and antimicrobial activity even at field realistic concentrations. Clothianidin had an effect on these immune parameters only at higher than field realistic concentrations (50–200 μg/l). These results suggest that neonicotinoids affect the individual immunocompetence of honey bees, possibly leading to an impaired disease resistance capacity.     

Serum levels of irisin in gestational diabetes mellitus during pregnancy and after delivery

ObjectiveIrisin has recently been introduced as a novel an exercise-inducible myokine which improves glucose metabolism in mice. However, regulation of circulating irisin in gestational diabetes mellitus (GDM) and in the peripartal period has not been assessed so far.MethodsCirculating irisin was quantified in 74 GDM patients and in 74 healthy, pregnant, gestational age-matched controls. In a subset of these patients (44 GDM, 41 controls), postpartum follow-up data were also available. In a second study population of 40 healthy women with singleton pregnancies undergoing elective Cesarean section, irisin was assessed in maternal serum before and within 24 h after delivery, as well as in umbilical cord blood and in placental tissue.ResultsIn the first study population, median [interquartile range] irisin levels were significantly higher in GDM patients as compared to controls after delivery (previous GDM: 446.3 [146.9] μg/l; controls: 378.0 [111.4] μg/l) but not during pregnancy (GDM: 482.1 [132.1] μg/l; controls: 466.6 [178.0] μg/l). Interestingly, fasting insulin (FI) was independently and positively associated with serum irisin in multivariate analysis during pregnancy. In agreement with these findings, relative changes (ratio) of FI independently and positively predicted relative changes of irisin (ratio) in the second study population.ConclusionsThe myokine irisin is independently associated with FI in pregnancy. The physiological significance of these findings needs to be assessed in future experiments.     

Ectoine improves yield of biodiesel catalyzed by immobilized lipase

To improve the production of biodiesel by enzymatic conversion of triglycerides in cottonseed oil, compatible solutes were added to the solvent-free methanolysis system to prevent competitive methanol inhibition on the immobilized lipase (Novozym^® 435). The results indicated that the addition of ectoine increased biodiesel synthesis using a three-step methanol addition process. The concentration of methyl ester (ME) reached a maximum of 95.0% in the presence of 1.1 mmol/l ectoine, an increase of 20.9% compared to that in the absence of ectoine. On the other hand, excess ectoine decreased the ME concentration. Ectoine was also shown to enhance reuse of the immobilized lipase, significantly improving ME concentrations in each recycling test. Total concentrations of ME with added ectoine were about 1.5 times that without ectoine during five recycling tests (molar ratio of cottonseed oil to methanol, 1:4). Enzymatic reaction kinetics showed, in the concentration ranges of 0.8–1.14 mol/l and 0.03–8 mol/l for triglyceride and methanol, respectively, that ectoine had no effect on the initial reaction rates when methanol concentrations were below 0.5 mol/l. When methanol concentration exceeded 0.5 mol/l, the addition of 0.8 mmol/l ectoine increased the initial reaction rates, and the lipase exhibited a lower affinity for methanol and higher affinity for triglyceride (kinetic parameters of K_mA increase, K_mTG decrease). However, the initial reaction rates decreased significantly when 8 mmol/l ectoine was added, with the lipase having higher affinity for methanol and lower affinity for triglyceride (K_mA decrease, K_mTG increase). The supplementation of ectoine provided a new method for the purpose of improving yield of biodiesel catalyzed by enzyme.     

Variation in polyphenolic profile and in vitro antioxidant activity of eastern teaberry (Gaultheria procumbens L.) leaves following foliar development

Seasonal dynamics in the polyphenolic composition, antioxidant activity, and their relationships during plant development were evaluated for eastern teaberry (Gaultheria procumbens L.) leaves, a traditional herbal medicine of North American natives. With the complementary UHPLC-PDA-ESI-MS³, HPLC-PDA-fingerprint, Folin-Ciocalteau, and n-butanol/HCl assays of methanol-water (75:25, v/v) extracts, the dried leaf samples harvested monthly across the growing season under Polish climate conditions were found rich in structurally diverse polyphenols (149.2–210.7 mg/g DW) including the dominating salicylates (64.6–107.5 mg/g DW), proanthocyanidins (53.0–66.8 mg/g DW), and flavonoids (17.3–25.3 mg/g DW), and the accompanying chlorogenic acid isomers (2.4–4.4 mg/g DW) and simple phenolic acids (0.9–1.1 mg/g DW). Among 28 detected analytes, gaultherin (64.6–107.5 mg/g DW), miquelianin (14.6–21.1 mg/g DW), procyanidin A-type trimer (5.5–9.5 mg/g DW), and (–)-epicatechin (5.8–7.8 mg/g DW) were the most abundant. The phenolic levels and antioxidant activity parameters in the DPPH (EC₅₀, 15.0–18.2 μg DW/mL; 0.95–1.16 mmol Trolox equivalents/g DW) and FRAP (2.3–3.4 mmol Fe ²⁺/g DW; 0.86–1.26 mmol Trolox equivalents/g DW) assays showed parallel seasonal trends with maxima in September and October. As the subsequent correlation studies confirmed the determinative impact of polyphenols on the leaf antioxidant activity and its seasonal fluctuations, the Fall season could be recommended as optimal for harvesting the plant material for medicinal purposes and cost-effective production of natural health products.     

Effects of intermittent watering on water balance and feed intake in male Ethiopian Somali goats

The objective of this study was to investigate the regulation of water balance and dry matter (DM) intake of male Ethiopian Somali goats. Twenty-eight goats were grouped into four treatments; goats watered every day (W1), every 2nd day (W2), every 3rd day (W3) and every 4th day (W4) for a total period of 72 days. Plasma sodium and potassium were analyzed by flame photometry, plasma osmolality by freezing point depression, plasma protein concentration by TS refractometry, and the plasma vasopressin concentration by radioimmunoassay. Goats were fed grass hay ad libitum and were given 200 g of concentrates daily. The calculated daily water intake (ml/kg BW^0.75) of W1, W2, W3 and W4 was 128 ± 4, 86 ± 4, 88 ± 4 and 78 ± 4, respectively, (P < 0.001, W1 versus the other groups). Calculated total concentrates intake did not change, whereas Group W2 and W4 consumed less hay on days of water deprivation (W1 versus W2, P < 0.01; W1 versus W4, P < 0.001). Plasma sodium concentration of W1 was 143 ± 1 mmol/l. Intermittent watering increased plasma sodium concentration from 140 or 141 ± 1 mmol/l to 145 or 146 ± 1 mmol/l every 2nd day in group W2. In W3, it increased from 139 ± 1 to 146 ± 1 mmol/l and in W4 from 137 ± 1 to 149 ± 1 mmol/l (P < 0.001 for all) on days after watering compared to last day of water deprivation, respectively. Corresponding changes were observed in plasma osmolality. Total plasma protein concentration increased only in W3 from 65 ± 1 to 67 ± 1 mmol/l on day 3 (P < 0.01) and in W4 from 65 ± 1 to 67 and 68 ± 1 mmol/l on days 3 and 4, respectively (P < 0.05). The plasma vasopressin concentration of W1 was close to 0.80 ± 0.28 pmol/l on all days, but it increased on dehydration days in W2 (P < 0.05). In W3, the vasopressin concentration increased from 0.95 or 1.19 pmol/l the day after watering to 3.11 ± 0.28 pmol/l on day 3 (P < 0.01). In W4, it was 0.82 ± 0.31 pmol/l the day after watering and reached 6.06 ± 0.31 pmol/l on day 4 (P < 0.001). We conclude that male Ethiopian Somali goats can be watered once every 3rd days, and that longer interval is not recommended because it demands marked mobilization of water saving mechanisms and causes decreased DM intake.     

Determination of d-saccharic acid-1,4-lactone from brewed kombucha broth by high-performance capillary electrophoresis

Kombucha is a health tonic. d-Saccharic acid-1,4-lactone (DSL), a component of kombucha, inhibits the activity of glucuronidase, an enzyme indirectly related with cancers. To date, there is no efficient method to determine the content of DSL in kombucha samples. In this paper, we report a rapid and simple method for the separation and determination of DSL in kombucha samples, using the high-performance capillary electrophoresis (HPCE) method with diode array detection (DAD). With optimized conditions, DSL can be separated in a 50 cm length capillary at a separation voltage of 20 kV in 40 mmol/L borax buffer (pH 6.5) containing 30 mmol/L SDS and 15% methanol (v/v). Quantitative evaluation of DSL was determined by ultraviolet absorption at λ = 190 nm. The relationship between the peak areas and the DSL concentrations, in a specified working range with linear response, was determined by first-order polynomial regression over the range 50–1500 μg/mL with a detection limit of 17.5 μg/mL. Our method demonstrated excellent reproducibility and accuracy with relative standard deviations (RSD) of less than 5% DSL content (n = 5). This is the first report to determine DSL by HPCE. We have successfully applied this method to determine DSL in kombucha samples in various fermented conditions.     

Haematological and biochemical parameters and tissue accumulations of cadmium in Oreochromis niloticus exposed to various concentrations of cadmium chloride

Oreochromis niloticus, weighing 36.45 ± 1.12 g were exposed to 10%, 20% and 30% of the LC₅₀ of CdCl₂ which represents treatments (T1)1.68, (T2)3.36 and (T3)5.03 mg/l, respectively, for a period of 10, 20 and 30 days. It was found that, compared to a control group reading of 0.19 ± 0.03 μg/g dry weight, accumulation of Cd in the gills was significantly (p < 0.05) increased in samples ranging between 7.64 ± 0.86 and 61.73 ± 0.82 μg/g dry weight from T1 at 10 days to T3 at 30 days. The accumulation of Cd in the liver, meanwhile, was also observed to significantly increase (p < 0.05) with increasing time and concentrations with results ranging between 3.21 ± 0.12 and 181.61 ± 1.32 compared to the control group results of 0.29 ± 0.04 μg/g dry weight. Although muscles exhibited lower levels of accumulation than the gills and liver they still showed the same pattern of increase compared to the control group, with a significant difference ranging between 0.32 ± 0.02 and 2.16 ± 0.08 compared to the control group results of 0.03 ± 0.001 μg/g dry weight. Also, haematological parameters such as red blood cells (RBCs), haemoglobin (Hb) and haematocrit (Hct) were reduced in fish exposed to Cd at all periods, with significant differences (p < 0.05). Plasma glucose concentration showed a significant increase. Total protein levels of fish showed a significant reduction (p > 0.05) for all exposed treatments. Also, the total lipid level increased significantly as fish were exposed to increasing cadmium concentrations, compared to control fish. Finally, the activities of aspartate aminotransferase (AST IU/l) and alanine aminotransferase (ALT IU/l) showed a significant increase (p < 0.05) with increasing time and concentrations.