Resazurin-based 96-well plate microdilution method for the determination of minimum inhibitory concentration of biosurfactants

Objectives

To develop and validate a microdilution method for measuring the minimum inhibitory concentration (MIC) of biosurfactants.

Results

A standardized microdilution method including resazurin dye has been developed for measuring the MIC of biosurfactants and its validity was established through the replication of tetracycline and gentamicin MIC determination with standard bacterial strains.

Conclusion

This new method allows the generation of accurate MIC measurements, whilst overcoming critical issues related to colour and solubility which may interfere with growth measurements for many types of biosurfactant extracts.

     

Conservation of severely fragmented populations: lessons from the transformation of uncoordinated reintroductions of cheetahs (<Emphasis Type="Italic">Acinonyx jubatus</Emphasis>) into a managed metapopulation with self-sustained growth

We document and evaluate the use of metapopulation management to conserve a declining population of 217 cheetahs in 40 subpopulations. Metapopulation management resembles a natural metapopulation, but dispersal success, demographic rescue effects and genetic viability are enhanced by moving suitable individuals to selected habitat fragments. Unfortunately, history and results of metapopulation management are rarely published. Cheetahs, extirpated from 85% of South Africa, were reintroduced from Namibian and South African ranches into fenced reserves. During 1965–2009 343 cheetahs were reintroduced, yet reserves held only 289 in 2009. Then translocations of free-roaming cheetahs were halted, and numbers dropped to 217 on 40 reserves by 2012. A metapopulation project was launched, and key conservation problems indentified from interviews and records. Thirty-five percent of reserves had no breeding cheetahs, 13% were inbreeding, fence quality was erratic, 3% of cheetahs were sold into captivity annually, and 28% of cheetah mortalities were anthropogenic. Lions accounted for 31% of mortality, perhaps elevated by lion-inexperienced cheetahs and high lion densities. These problems were addressed, and cheetahs were translocated between reserves. Although the median reserve size was only 125 km² holding four cheetahs, and 80% of reserves were privately owned, in 6 years the metapopulation grew by 51% to 328 cheetahs on 51 reserves, while genetic diversity was managed and monitored. Thus, using metapopulation management, low density species and associated key processes, including carnivory or mega-herbivory, can be conserved in relatively small reserves in regions with dense human populations precluding  natural gene flow.     

The evolutionary dynamics of the lion Panthera leo revealed by host and viral population genomics

The lion Panthera leo is one of the world's most charismatic carnivores and is one of Africa's key predators. Here, we used a large dataset from 357 lions comprehending 1.13 megabases of sequence data and genotypes from 22 microsatellite loci to characterize its recent evolutionary history. Patterns of molecular genetic variation in multiple maternal (mtDNA), paternal (Y-chromosome), and biparental nuclear (nDNA) genetic markers were compared with patterns of sequence and subtype variation of the lion feline immunodeficiency virus (FIV(Ple)), a lentivirus analogous to human immunodeficiency virus (HIV). In spite of the ability of lions to disperse long distances, patterns of lion genetic diversity suggest substantial population subdivision (mtDNA Phi(ST) = 0.92; nDNA F(ST) = 0.18), and reduced gene flow, which, along with large differences in sero-prevalence of six distinct FIV(Ple) subtypes among lion populations, refute the hypothesis that African lions consist of a single panmictic population. Our results suggest that extant lion populations derive from several Pleistocene refugia in East and Southern Africa ( approximately 324,000-169,000 years ago), which expanded during the Late Pleistocene ( approximately 100,000 years ago) into Central and North Africa and into Asia. During the Pleistocene/Holocene transition ( approximately 14,000-7,000 years), another expansion occurred from southern refugia northwards towards East Africa, causing population interbreeding. In particular, lion and FIV(Ple) variation affirms that the large, well-studied lion population occupying the greater Serengeti Ecosystem is derived from three distinct populations that admixed recently.     

Fenced and Fragmented: Conservation Value of Managed Metapopulations

Population fragmentation is threatening biodiversity worldwide. Species that once roamed vast areas are increasingly being conserved in small, isolated areas. Modern management approaches must adapt to ensure the continued survival and conservation value of these populations. In South Africa, a managed metapopulation approach has been adopted for several large carnivore species, all protected in isolated, relatively small, reserves that are fenced. As far as possible these approaches are based on natural metapopulation structures. In this network, over the past 25 years, African lions (Panthera leo) were reintroduced into 44 fenced reserves with little attention given to maintaining genetic diversity. To examine the situation, we investigated the current genetic provenance and diversity of these lions. We found that overall genetic diversity was similar to that in a large national park, and included a mixture of four different southern African evolutionarily significant units (ESUs). This mixing of ESUs, while not ideal, provides a unique opportunity to study the impact of mixing ESUs over the long term. We propose a strategic managed metapopulation plan to ensure the maintenance of genetic diversity and improve the long-term conservation value of these lions. This managed metapopulation approach could be applied to other species under similar ecological constraints around the globe.     

Synchronization of estrus in beef heifers using either melengesterol acetate (MGA)/prostaglandin or MGA/Select Synch

The objective of this study was to evaluate synchronization, conception, and pregnancy rates of yearling beef heifers synchronized with either the Select Synch protocol preceded by 7 days of MGA feeding (MGA/Select Synch) or the traditional MGA/PGF protocol. Heifers in the MGA/Select Synch group (n = 402) were fed MGA (0.5 mg/day/head) for 7 days, received an injection of GnRH (100 microg) the day following the last MGA feeding and an injection of PGF (25 mg) 7 days after GnRH. Heifers in the MGA/PGF group (n = 394) received MGA (0.5 mg/day/head) for 14 days, followed by an injection of PGF (25 mg) 17 days later. Synchronization rates tended (P = 0.08) to be higher for the MGA/Select Synch (82%) compared to the MGA/PGF (77%)-treated heifers. Conception and pregnancy rates to AI were similar (P > 0.10), 57 and 46% for the MGA/Select Synch heifers and 61 and 47% for the MGA/PGF heifers, respectively. Mean estrous response (h) was earlier (P < 0.05) for the MGA/Select Synch versus MGA/PGF treatment, 56 versus 61 h post-PGF treatment, respectively. In summary, short-term (7 days) MGA feeding preceding the Select Synch protocol produced similar synchronization, conception, and pregnancy rates as the traditional MGA/PGF protocol.     

Changes in lion (Panthera leo) home range size in Waza National Park,Cameroon

The spatial ecology of Africa lions (Panthera leo) was studied from 2007 to 2009 in Waza National Park, Cameroon, by equipping individual lions with GPS/VHF radio-collars. Mean home range estimates using 100% minimum convex polygons (MCP) and 95% kernel-density estimation (KDE) were respectively 1015 km² and 641 km². The lions spent a considerable amount of time out of the park during the study period (21%), resulting in significantly larger wet season home ranges than in the hot dry season when they were largely within the park. Time spent outside of the park coincided with increased livestock predation, especially by males. The seasonal variation observed in home range appeared to be mainly due to prey dispersal, flooding and migrating livestock. Mean home range size was observed to have increased by 58.6% within the last decade. This observed increase in home range could possibly be attributed to recent declines in wild prey abundance and also, may be indicative of a trend of general degradation of the park due to intense human pressure. The change observed in lions’ ranging behaviour was remarkable, with lions crossing the highway parallel to the park to the Cameroon-Nigerian borders. Measures to restore the integrity of the park are urgently needed, which could include the construction of a partial fence along the western boundary of the park to prevent lions moving across the parallel highway.     

Trade-offs of predation and foraging explain sexual segregation in African buffalo

1. Many studies have investigated why males and females segregate spatially in sexually dimorphic species. These studies have focused primarily on temperate zone ungulates in areas lacking intact predator communities, and few have directly assessed predation rates in different social environments. 2. Data on the movement, social affiliation, mortality and foraging of radio-collared African buffalo (Syncerus caffer) were collected from 2001-06 in the Kruger National Park, South Africa. 3. The vast majority of mortality events were due to lion (Panthera leo) predation, and the mortality hazard associated with being an adult male buffalo in a male-only 'bachelor' group was almost four times higher than for adult females in mixed herds. The mortality rates of adult males and females within mixed herds were not statistically different. Mortality sites of male and female buffalo were in areas of low visibility similar to those used by bachelor groups, while mixed herds tended to use more open habitats. 4. Males in bachelor groups ate similar or higher quality food (as indexed by percentage faecal nitrogen), and moved almost a third less distance per day compared with mixed herds. As a result, males in bachelor groups gained more body condition than did males in breeding herds. 5. Recent comparative analyses suggest the activity-budget hypothesis as a common underlying cause of social segregation. However, our intensive study, in an area with an intact predator community showed that male and female buffalo segregated by habitat and supported the predation-risk hypothesis. Male African buffalo appear to trade increased predation risk for additional energy gains in bachelor groups, which presumably leads to increased reproductive success.     

Conserving large carnivores: dollars and fence

Conservationists often advocate for landscape approaches to wildlife management while others argue for physical separation between protected species and human communities, but direct empirical comparisons of these alternatives are scarce. We relate African lion population densities and population trends to contrasting management practices across 42 sites in 11 countries. Lion populations in fenced reserves are significantly closer to their estimated carrying capacities than unfenced populations. Whereas fenced reserves can maintain lions at 80% of their potential densities on annual management budgets of $500 km⁻², unfenced populations require budgets in excess of $2000 km⁻² to attain half their potential densities. Lions in fenced reserves are primarily limited by density dependence, but lions in unfenced reserves are highly sensitive to human population densities in surrounding communities, and unfenced populations are frequently subjected to density‐independent factors. Nearly half the unfenced lion populations may decline to near extinction over the next 20–40 years.     

Genetic diversity,evolutionary history and implications for conservation of the lion (Panthera leo) in West and Central Africa

Aim In recent decades there has been a marked decline in the numbers of African lions (Panthera leo), especially in West Africa where the species is regionally endangered. Based on the climatological history of western Africa, we hypothesize that West and Central African lions have a unique evolutionary history, which is reflected by their genetic makeup. Location Sub‐Saharan Africa and India, with special focus on West and Central Africa. Method In this study 126 samples, throughout the lion’s complete geographic range, were subjected to phylogenetic analyses. DNA sequences of a mitochondrial region, containing cytochrome b, tRNAPro, tRNAThr and the left part of the control region, were analysed. Results Bayesian, maximum likelihood and maximum parsimony analyses consistently showed a distinction between lions from West and Central Africa and lions from southern and East Africa. West and Central African lions are more closely related to Asiatic lions than to the southern and East African lions. This can be explained by a Pleistocene extinction and subsequent recolonization of West Africa from refugia in the Middle East. This is further supported by the fact that the West and Central African clade shows relatively little genetic diversity and is therefore thought to be an evolutionarily young clade. Main conclusions The taxonomic division between an African and an Asian subspecies does not fully reflect the overall genetic diversity within lions. In order to conserve genetic diversity within the species, genetically distinct lineages should be prioritized. Understanding the geographic pattern of genetic diversity is key to developing conservation strategies, both for in situ management and for breeding of captive stocks.