Alcohol diluent provides the optimal formulation for calcium chloride non-surgical sterilization in dogs

Background

Surgical castration is widely used to sterilize male dogs, but has significant impacts on time to perform the operation, recovery of the animals as well as cost, which can limit population control programs. Previous research has shown intratesticular injection of calcium chloride dihydrate (CaCl₂) in saline to be a promising alternative to surgery. However, long-term azoospermia was not maintained at dosages low enough to avoid side effects. In the search for an optimized formulation, the current investigation is the first study on long-term sterilization effects of intratesticular injection of CaCl₂ in either lidocaine solution or alcohol in dogs. CaCl₂ at 20% concentration in lidocaine solution or alcohol was administered via intratesticular injection to groups of 21 dogs each. The treated animals were examined at 2, 6, and 12 months for sperm production, blood levels of testosterone, and side effects; at time zero and 12 months for testicular size and semen volume. The experimentally treated animals were compared to a control group receiving saline injection only.

Results

Testicles of dogs treated with CaCl₂ in either diluent significantly decreased in size. After administration of CaCl₂ in lidocaine solution, sterility was achieved for at least 12 months in 75% of treated dogs. However, optimal long-term contraceptive effectiveness was achieved with CaCl₂ in alcohol, which resulted in azoospermia over the 12-month study period. Testosterone levels significantly decreased following treatment with CaCl₂, and sexual activity disappeared. Although testosterone returned to baseline levels by 12 months for the group treated with CaCl₂ in lidocaine, dogs injected with CaCl₂ in alcohol had a 63.6% drop in testosterone level, which remained at the low end of physiological range throughout the study. No adverse effects were noted.

Conclusions

A single, bilateral intratesticular injection of 20% CaCl₂ in 95% ethanol was a reliable method for induction of sterilization in 18–28 kg male dogs in this study. The approach showed long-term efficacy and reduced sexual behavior. This chemical method of sterilization might provide an effective, efficient alternative to surgical castration that can have positive impacts on dog welfare.     

Evaluation of the effectiveness of Duddingtonia flagrans and Monacrosporium thaumasium in the biological control of gastrointestinal nematodes in female bovines bred in the semiarid region

Brazil has a herd of 212 million cattle and 171 million hectares of pastures that produce approximately 96 % of Brazilian beef. The Brazilian production system enables animal infection by endoparasites, which are considered one of the main obstacles for the development of this industry and are responsible for considerable economic losses. The control of parasitic diseases is performed via the administration of antiparasitic drugs, but they leave residues of the products in the treated animal, affect non-target organisms and select resistant strains of the parasites. The species D. flagrans and M. thaumasium are promising and sustainable alternatives for controlling gastrointestinal helminths of ruminants and other herbivores. In this study, we evaluated the efficacy of isolates of these species, formulated in a sodium alginate matrix and administered twice a week, to reduce the number of environmental infective larvae of gastrointestinal nematodes that affect prepubescent zebu females. The treated animals presented fewer eggs and a lower number of infective larvae per gram of faeces (p?<?0.05). The pastures occupied by treated animals showed a statistically significant reduction (p?<?0.05) of the number of L₃ and, furthermore, the genera Cooperia sp., Haemonchus sp., and Oesophagostomum sp. were the most prevalent. The average weight of the animals did not differ statistically (p?>?0.05) among the treated and control groups. The use of sodium alginate pellets as vehicle for delivery of the fungus mycelia D. flagrans (isolate AC001) and M. thaumasium (isolate NF34A) proved effective in controlling trichostrongylids in prepubescent cows bred in the semi-arid region, with an effective reduction in the number of infective larvae in the pastures.     

Use of Thermography Techniques in Equines: Principles and Applications

This review of the use of thermographic technique in equines introduces the principles upon which infrared radiation and thermoregulatory physiology are based and describes the instrumentation used and its practical use. The advantage of this imaging technique is that it is a noninvasive thermographic examination, both from an operational (the animal and the operator) and health (no penetrating radiation is used) standpoint. Advantages and disadvantages of this technique, equine applications, and physiological assessments are discussed.     

Space–time patterns of co‐variation of biodiversity and primary production in phytoplankton guilds of coastal marine environments

• 1. The relevance of biodiversity to ecosystem processes is a major topic in ecology. Here, we analyse the relationship between biodiversity and productivity of the nano‐ and micro‐phytoplankton guilds in coastal marine ecosystems.
• 2. The patterns of variation of species richness, diversity and primary productivity (as ¹⁴C assimilation) were studied in two marine areas: a eutrophic–mesotrophic area beside the River Po delta (northern Adriatic) and an oligotrophic area around the Salento peninsula (southern Adriatic–Ionian). The study was carried out at 23 sites in the northern area and at 45 sites in the southern area. Sites were arranged on expected spatial and temporal gradients of primary productivity variation, according to distance from the coast, optical depths and seasonal period.
• 3. 167 taxa were identified in the northern area and 153 taxa in the southern area. In both areas, the taxonomic composition of the nano‐ and micro‐phytoplankton guilds exhibited greater temporal than spatial variation. The latter was much higher in the southern area than in the northern area (average dissimilarity between stations being 70.7±0.8% and 44.7±4.2% respectively).
• 4. Primary productivity varied in space and time on the gradients considered. Phytoplankton species richness and diversity exhibited significant patterns of variation in space and time; overall, these were inversely related to the primary productivity patterns in the northern area, whereas they were directly related in the southern area.
• 5. The small individual size and the high turnover rate of phytoplankton are likely to underlie the observed relationships, which emphasized a threshold response to nutrient enrichment in agreement with the ‘paradox of enrichment’. Under resource enrichment conditions, the high turnover of producers leads to hierarchical partitioning of the available resources with an increasing dominance of a few species. Therefore, the relationship observed here seems likely to be explained by the complementarity hypothesis.

Copyright © 2003 John Wiley & Sons, Ltd.     

Drying-rewetting cycles and γ-irradiation effects on enzyme activities of distinct layers from a Quercus ilex L. litter

In a Mediterranean climate, water stress is one of the principal constraints on proper forest ecosystem functioning. Drought influences rates of organic matter degradation by affecting microbial growth and enzyme activities. The objectives of this study were: (i) to evaluate the effect of repeated drying-rewetting cycles on cellulase, alkaline phosphatase and fluorescein diacetate (FDA) hydrolase activities of three distinct Quercus ilex L. litter layers, and (ii) to investigate the effect of these cycles on γ-irradiated litters in order to distinguish the abiotic influence on the fluctuations observed. Results, for all three layers, showed high correlations between litter water content and enzyme activities. Under mesocosm conditions, and using non-sterilized litter samples, cellulase, alkaline phosphatase, and FDA activities significantly decreased or increased during drying or rewetting cycles respectively. Significant differences were also found when evaluating the effect of litter depth on enzyme activities, the intermediate depth (OLv layer) generally being the most active. For γ-sterilized samples, FDA activity still fluctuated with drying-rewetting cycles. Assays showed that pre-humidification of γ-irradiated litter increased FDA activity two-fold in the first 30 min. All these results have shown that, following drying-rewetting cycles, some of the fluctuations occur independently of microbial growth, suggesting abiotic interactions, such as desorption, in combination with both solvatation status and conformational changes of enzymes.     

Studies on the constituents of a Brazilian folk infusion. Isolation and structure elucidation of new triterpene saponins from Ilex amara leaves.

The isolation of three new triterpene saponins 3beta-O-beta-D-glucopyranosyl-(1-->3)-alpha-L-2-O-acetylarabinopyranosylolean-12-en-28-oic acid 28-O-beta-D-glucopyranosyl ester (2), 3beta-O-beta-D-glucopyranosyl-(1-->2)-alpha-L-O-arabinopyranosylurs-12-en-28-oic acid (3), and 3beta-O-beta-D-glucopyranosyl-(1-->2)-beta-D-O-galactopyranosylurs-12-en-28-oic acid (4) together with five known saponins and one flavonoid glycoside from the aqueous infusion of Ilex amara (Vellozo) Loes. leaves is reported. All structures were elucidated by spectroscopic methods, including the concerted application of one-dimensional (1H, TOCSY, 13C, and 13C DEPT NMR) and two-dimensional NMR techniques (DQF-COSY, HSQC, and HMBC).     

Quantification of the biocontrol agent Pseudomonas fluorescens Pf153 in soil using a quantitative competitive PCR assay unaffected by variability in cell lysis- and DNA-extraction efficiency

Although often neglected, variability in cell lysis efficiency and DNA extraction yield represents the major hurdles of any polymerase chain reaction (PCR)-based quantification protocol in soil and other natural environments. In this study we developed a technique that minimizes the effects of these constraints, providing at the same time a reliable internal control to distinguish between PCR-inhibition and negative results. We used Pseudomonas fluorescens Pf153, a root-colonizing bacterium that shows biocontrol activity against tobacco and cucumber black root rot, as the target organism for PCR quantification. Prior to DNA extraction, the genetically engineered, cognate reference strain P. fluorescens CHA0/c2 was inoculated in a reference soil. CHA0/c2 in the reference soil and Pf153 in the soil sample were lysed in parallel and afterward the lysates were mixed in known proportions. CHA0/c2 carries the plasmid pME6031-cmp2 that contains an allelic variant (competitor) of the Pf153 specific sequence Pf153_2. In a quantitative competitive PCR (QC-PCR) assay the competitor allows the quantification of the target strain down to 0.66 Pf153 CFU/mg soil. Processing the reference strain in the same way as Pf153 enables the exact quantification of the target strain in biocontrol assays performed in natural soil, overcoming differences in DNA extraction efficiency and PCR amplification from different soil environments. This technique is easily adaptable to other Pseudomonas strains simply by replacing the competitor used here with one derived from a SCAR-marker which is specific for the strain of choice.     

Pre-unfolding resonant oscillations of single green fluorescent protein molecules

Fluorescence spectroscopy of a green fluorescent protein mutant at single-molecule resolution has revealed a remarkable oscillatory behavior that can also be driven by applied fields. We show that immediately before unfolding, several periodic oscillations among the chemical substates of the protein chromophore occur. We also show that applied alternating electric or acoustic fields, when tuned to the protein characteristic frequencies, give rise to strong resonance effects.