Identification of tissue hypoxia in the livers of ascitic and hypoxia‐induced broilers using trypan blue

1. The perfusion of livers with the vital dye trypan blue was performed to test for evidence of tissue hypoxia in 3 groups of young broiler chickens, namely, ascitic, hypoxia‐induced and controls.

2. Hepatocytes that stained with trypan blue were considered to be dead or dying before fixation and represented damaged cells.

3. The proportion of trypan blue‐stained hepatocytes in the livers of ascitic birds was slightly less than half that observed in the hypoxia‐induced birds but significandy more than the proportion of stained cells observed in control birds.

4. Liver damage in the ascitic birds was also assessed biochemically by an altered enzyme profile.

5. The study demonstrated that increased trypan blue uptake in the livers of ascitic birds reared at sea‐level may be the consequence of hypoxia stress caused by reduced oxygen utilisation.     

Some currently available insecticides and their comparative efficacy on louse-infested, long-woolled sheep

The comparative efficacy of 13 of the sheep dips currently registered in New Zealand was investigated using sheep infested with the louse Bovicola ovis and carrying wool which was about 10 cm long at the shoulder. With the exception of one synthetic pyrethroid pour-on formulation, all products were able to effect a significant reduction in louse populations, relative to untreated controls, for 37 days after treatment. Only four products proved capable of eradicating lice and preventing their re-establishment up to 37 days after treatment. Variations in manufacturers' recommendations relating to the length of wool at dipping, and mode of application of dips are discussed in relation to the results.     

An evaluation of two cypermethrin-based pour-on formulations on sheep infested with the biting louse, Bovicola ovis

Two synthetic pyrethroid (cypermethrin) based pour-on insecticide formulations, with high cis/trans isomer ratios (80:20) but differing in their respective active ingredient concentrations and solvent component(s), were applied to sheep infested with the biting-louse, Bovicola ovis. All treated sheep were penned with louse-infested sheep 9,12 and 15 weeks after the insecticide was applied. The 2% cypermethrin formulation achieved a higher level of control than the 1.25% cypermethrin formulation at each challenge interval when applied 12 weeks after shearing. The 2% cypermethrin formulation provided 97-100% control of lice from 4 to 16 weeks after application on sheep shorn 6 or 12 weeks prior to treatment. The 1.25% cypermethrin formulation provided 85% control of lice 4 weeks after application on sheep shorn 12 weeks prior to treatment, the level of control increasing to a maximum of 100% by week 9, and declining thereafter. The 2% cypermethrin formulation may provide a better level of control in long-woolled sheep than 1.25% cypermethrin, by compensating for the diluent effect of lipid.     

The Impact of Natural Deep Eutectic Solvents and Extraction Method on the Co-Extraction of Trace Metals from Fucus vesiculosus

In recent years, natural deep eutectic solvents (NADES) have been widely investigated for the extraction of food and medicinal plants as well as seaweeds. However, the ability of NADES for trace elements co-extraction from natural sources is not well investigated. The aim of this study was to investigate the ability of common NADES for trace elements co-extraction from Fucus vesiculosus. All of the tested NADES did not recover As and Co (concentration <LOQ). Moreover, all of the tested NADES provided a low recovery (<9%) of Ba, Ca, Fe, Mg, Mn, Sr, and Zn. The method of extraction had not shown a statistically significant effect on the co-extraction of all elements (excluding Ba and Ca). In contrast, the water content in NADES was significantly affected on the recovery of Ba, Ca, Mg, Mn, Sr, and Zn. The recovery of Al and Cr was relatively high and considerably varied (from 1.5 to 59.9%). NADES comprising lactic acid:glucose:H₂O (5:3:1) provided the lowest contents of all elements, and the highest extracted amounts were obtained employing water contents of 60–80%. The calculated daily intake of all the elements contained in NADES extracts were less than the daily dose risk estimators. The hazard quotients, hazard indexes, and carcinogenic risk calculated for all trace elements and their combination were considerably less than 1. This evidences no health risk, and carcinogenic risk after topical application of all studied NADES. For the first time, the results of the current study demonstrated that NADES extracts of F. vesiculosus contain a lower amount of trace metals and are safer than the extracts obtained with water and 70% acetone. This indicates a significant advantage for NADES compared with the other solvents.     

Mechanisms of persistence of low numbers of bacteria preyed upon by protozoa

Tetrahymena pyriformis cultures were maintained when transferred serially in solutions containing 10⁵ to 10⁷Klebsiella pneumoniae cells.ml^?1, bacterial numbers that were observed to persist in the presence of protozoa. The number of cells of one strain of K. pneumoniae surviving predation in solution was essentially the same in the absence of an alternative prey as in the presence of a second K. pneumoniae strain. Toxins deleterious to protozoa did not appear as the animal consumed the bacteria. T. pyriformis reduced the abundance of Escherichia coli from about 10⁸ to 10⁶.ml^?1. but the latter number persisted for 15 days; however, in solutions containing chloramphenicol, the abundance of E. coli fell to 590 cells. ml^?1 in 15 days. In solutions containing the antibiotic, T. pyriformis reduced the Rhizobium sp. population from more than 10⁶ to less than 10³ cells in 10 days and K. pneumoniae from more than 10⁸.ml^?1 to zero in 18 days. An appreciable decline in abundance of these bacteria did not occur in the antibiotic-amended liquid free of protozoa. T. pyriformis did not greatly reduce Rhizobium sp. numbers when both were added to irradiated soil, but the predator caused the bacterial population to decline from 4 × 10⁸ to fewer than 10⁵.g^?1 in 16 days in chloramphenicol-treated soil. Colpoda sp. inoculated with Rhizobium sp. into soil sterilized by autoclaving only reduced the prey abundance from 10⁹ to 10⁸.g^?1, but the protozoan caused the bacterial population to fall to about 100.g^?1 in 15 days in the presence of the antibiotic. The population of Rhizobium sp. added to nonsterile soil dropped from in excess of 10⁸ to 6 × 10⁶.g^?1 in 29 days. but it declined to 550. g^?1 in the same period when chloramphenicol was also introduced. It is concluded that the ability of these bacteria to maintain themselves in solution and in soil is governed by their capacity to reproduce and replace the cells consumed by predation.     

Formation of volatile organic products in soils under anaerobiosis—I: Metabolism of glucose

A simple trapping procedure for concentration of volatile organic compounds present in a head space and the subsequent analysis of the retained compounds by combined gas chromatographymass spectrometry is described. The procedure was used to study the organic metabolites evolved from various soils when they were amended with glucose and incubated under argon. A number of organic acids, alcohols, aldehydes and esters were evolved during a 4 week period. Less than 10 per cent of the added carbon was recovered as organic compounds, which was largely due to poor recovery of volatile organic acids from soil. Some implications of the observations to microbial ecology are discussed.