Multivariate calibration for the determination of total azadirachtin-related limonoids and simple terpenoids in neem extracts using vanillin assay

Two-component and multivariate calibration techniques were developed for the simultaneous quantification of total azadirachtin-related limonoids (AZRL) and simple terpenoids (ST) in neem extracts using vanillin assay. A mathematical modeling method was also developed to aid in the analysis of the spectra and to simplify the calculations. The mathematical models were used in a two-component calibration (using azadirachtin and limonene as standards) for samples containing mainly limonoids and terpenoids (such as neem seed kernel extracts). However, for the extracts from other parts of neem, such as neem leaf, a multivariate calibration was necessary to eliminate the possible interference from phenolics and other components in order to obtain the accurate content of AZRL and ST. It was demonstrated that the accuracy of the vanillin assay in predicting the content of azadirachtin in a model mixture containing limonene (25% w/w) can be improved from 50% overestimation to 95% accuracy using the two-component calibration, while predicting the content of limonene with 98% accuracy. Both calibration techniques were applied to estimate the content of AZRL and ST in different parts of the neem plant. The results of this study indicated that the relative content of limonoids was much higher than that of the terpenoids in all parts of the neem plant studied.     

Estimated prevalence of nematode parasitism among pet cats in the United States

OBJECTIVE: To estimate prevalences of roundworm, hookworm, and whipworm infections in pet cats in the United States and identify risk factors for parasitism. DESIGN: Retrospective period prevalence survey. STUDY POPULATION: 356,086 cats examined at 359 private veterinary hospitals during 2003. PROCEDURE: Electronic medical records were searched to identify cats for which fecal flotation tests had been performed and to determine proportions of test results positive for roundworms, hookworms, and whipworms. Potential risk factors for roundworm and hookworm infection were identified by means of multivariate logistic regression analysis. RESULTS: A total of 80,278 tests were performed on fecal samples from 66,819 cats. Calculated prevalences of roundworm, hookworm, and whipworm infection were 2.92%, 0.63%, and 0.031%, respectively. Age, reproductive status, breed, and season were significant risk factors for roundworm infection, with cats < 4 years old; sexually intact cats; mixed-breed cats; and cats examined during the summer, fall, or winter more likely to be infected. Age, reproductive status, and season were significant risk factors for hookworm infection, with cats < 1 year old, sexually intact cats, and cats examined during the summer more likely to be infected. Regional differences in prevalences of roundworm and hookworm infection were found. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that prevalences of nematode infections among pet cats in the United States may be lower than previously suspected on the basis of prevalences reported among cats in humane shelters and those reported in more geographically focused studies.     

Prevalence and risk factors for Giardia and coccidia species of pet cats in 2003-2004

Prevalence and risk factors for feline coccidia and Giardia species infections were estimated for cats visiting 434 Banfield hospitals in 40 states in 2003-2004. Evaluated were 631,021 cats making 1,456,712 office visits (encounters) and having 211,105 fecal examinations. The overall fecal prevalences of coccidia and Giardia species were 1.4% and 0.58%, respectively. Cats at increased risk of coccidia infection were under 4 years of age, intact, and seen during the summer, fall, and spring months compared to winter. Cats at increased risk of Giardia species infection were under 4 years of age. Those at decreased risk were mixed breed and seen during the summer, fall, and spring. The highest regional risk of coccidia and Giardia species infection was for cats in the East South Central region and Mountain region, respectively, compared to the South Pacific region.     

Returns and determinants of technical efficiency in small-scale Malabari goat production units in Kerala, India

A stochastic frontier production function was employed to measure technical efficiency and its determinants in smallholder Malabari goat production units in Kerala, India. Data were obtained from 100 goat farmers in northern Kerala, selected using multistage random sampling. The parameters of the stochastic frontier production function were estimated using the maximum likelihood method. Cost and return analysis showed that the major expenditure was feed and fodder, and veterinary expenses were secondary. The chief returns were the sale of live animals, milk and manure. Individual farm technical efficiency ranged from 0.34 to 0.97 with a mean of 0.88. The study found herd size (number of animal units) and centre (locality of farm) significantly affected technical efficiency, but sex of farmer, education, land size and family size did not. Technical efficiency decreased as herd size increased; half the units with five or more adult animals had technical efficiency below 60 %.     

Radical scavenging and reducing ability of tilapia (Oreochromis niloticus) protein hydrolysates

Enzymatically hydrolyzed fish protein hydrolysates could be used as a source of antioxidative nutraceuticals. In our current work, we have investigated alkali-solubilized tilapia ( Oreochromis niloticus) protein hydrolysates for their ability to scavenge reactive oxygen species (ROS) and for their reducing power. Tilapia protein isolate was prepared by an alkaline solubilization technique and used as a substrate for enzyme hydrolysis. Cryotin, protease A 'Amano' 2, protease N 'Amano', Neutrase and Flavourzyme, were used separately to determine their effectiveness in hydrolyzing tilapia protein isolate. ROS scavenging ability was quantified using an isoluminol enhanced chemiluminescent assay in the presence of a) hydrogen peroxide or b) mononuclear cells isolated from human blood. Ferric reducing antioxidant power (FRAP) and Trolox equivalent antioxidant capacity (TEAC) of the hydrolysates using 2, 2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) or 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), were also investigated. Results showed that, in general, the TEAC, FRAP values and ROS scavenging ability of the hydrolysates increased with an increase in the degree of hydrolysis. Among the different hydrolysates, those prepared using Cryotin were most effective and Amano A2 hydrolysates were least effective in scavenging ABTS*(+) and ROS generated by hydrogen peroxide. However, FRAP assay showed that hydrolysates prepared using Flavourzyme were most effective, and Amano N and Neutrase hydrolysates were least effective in reducing ferric ions. No significant difference was observed among the hydrolysates produced with different enzymes in their ability to scavenge ROS generated by phorbol myristate acetate stimulated mononuclear cells. These results shed light on the in vitro ROS scavenging ability of alkali solubilized tilapia protein hydrolysates, as well as potential nutraceutical use of these hydrolysates.     

Novel mechanism of modulating natural antioxidants in functional foods: involvement of plant growth promoting Rhizobacteria NRRL B-30488

The significance of plant growth-promoting rhizobacteria (PGPR) mediated increase in antioxidant potential in vegetables is yet unknown. The plant growth-promoting bacterium Bacillus lentimorbus NRRL B-30488 (B-30488) mediated induction of dietary antioxidant in vegetables ( Trigonella foenum-graecum, Lactuca sativa, Spinacia oleracea, and Daucus carota) and fruit ( Citrus sinensis) after minimal processing (fresh, boiled, and frozen) was tested by estimating the total phenol content, level of antioxidant enzymes, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide scavenging activities along with integral radical scavenging capacity by photochemiluminescence assay and inhibition of lipid peroxidation. Minimal processing of vegetables showed that T. foenum-graecum had the highest phenol content in B-30488-treated plants followed by L. sativa, D. carota, and S. oleracea. Thermally treated vegetables T. foenum-graecum (26-114.5 GAE microg mg (-1)) had an exceptionally high total phenolic content, followed by D. carota (25.27-101.32 GAE microg mg (-1)), L. sativa (23.22-101.10 GAE microg mg (-1)), and S. oleracea (21.87-87.57 GAE microg mg (-1)). Among the vegetables and fruit used in this study for enzymatic estimation, induction of antioxidant enzymes, namely, polyphenol oxidase (PPO), ascorbate peroxidase (APX), catalase (CAT), and superoxidase dismutase (SOD), was observed in edible parts of T. foenum-graecum, L. sativa, S. oleracea, and D. carota, after inoculation with B-30488. The scavenging capacity of the vegetables treated with B-30488 against DPPH and superoxide anion radical activity was found to be significantly high as compared to nontreated control. Mild food processing had no adverse effect on radical scavenging capacity. Photochemiluminescence also ascertains the above findings. The ability of the plant extracts to protect against lipid peroxidation and its ability to prevent oxidation of reduced glutathione (GSH) was measured in rat liver homogenate, and the results suggested that the inoculated plant exhibited better activity in all of the screened plants. Significant increases in shoot length, root length, and dry weight, averaging 164, 132, and 135% in T. foenum-graecum, 174, 141, and 156% in L. sativa, 129, 141, and 59%, in S. oleracea, and 125, 146, and 42% in D. carota, respectively, over untreated controls, were attained in greenhouse trials. To the best of the authors' knowledge, this is the first report of PGPR-mediated induction of antioxidant enzyme activity (PPO, APX, CAT, and SOD) along with the antioxidant activity of the extracts in both in vitro (DPPH radical scavenging and superoxide scavenging) and ex vivo conditions using the rat liver tissue (percent inhibition of lipid peroxidation and prevention of oxidation of GSH) and phenolic content. The results demonstrate the PGPR-mediated induction of antioxidant level in vegetables and fruit controls oxidative damage even after minimal processing and thus is indicative of its potential as a viable substitute of synthetic antioxidants.     

Origins and genetic legacy of Neolithic farmers and hunter-gatherers in Europe

The farming way of life originated in the Near East some 11,000 years ago and had reached most of the European continent 5000 years later. However, the impact of the agricultural revolution on demography and patterns of genomic variation in Europe remains unknown. We obtained 249 million base pairs of genomic DNA from ~5000-year-old remains of three hunter-gatherers and one farmer excavated in Scandinavia and find that the farmer is genetically most similar to extant southern Europeans, contrasting sharply to the hunter-gatherers, whose distinct genetic signature is most similar to that of extant northern Europeans. Our results suggest that migration from southern Europe catalyzed the spread of agriculture and that admixture in the wake of this expansion eventually shaped the genomic landscape of modern-day Europe.     

Risk analysis of non-native three-spot cichlid,Amphilophus trimaculatus, in the River Cauvery (India)

The three-spot cichlid Amphilophus trimaculatus (Günther) is an ornamental fish known to be invasive but has not been assessed for India. The present study confirmed this non-native species’ identity using conventional and molecular methods, as well as its occurrence in the River Cauvery, the first for this species in a lotic environment outside its native range. Gut content analysis indicated a diet of predominantly insects. The river's water quality and habitat were conducive for the species’ growth and reproduction. Using the Aquatic Species Invasiveness Screening Kit, A. trimaculatus was ranked as posing a “high” risk of being invasive in the River Cauvery, with climate match analysis showing 58% similarity with the species’ native range. Vector and pathway analysis found that A. trimaculatus was introduced to India via the aquarium trade, although consignment origin remains unknown, and the most likely means of entry into the river and its tributaries was the species’ escaping from holding facilities. Subsequent dispersal and impact of the species on native fauna are discussed. The study's outcomes demonstrate the importance of using risk analysis protocols to inform management of non-native species in India, with suggestions provided for immediate and long-term measures for the species’ management.