An Akania (Akaniaceae) inflorescence with associated pollen from the early Miocene of New Zealand.

PREMISE OF THE STUDY: An Akania-like inflorescence, including flowers with in situ pollen was recovered from the remarkable Konservat-Lagerstätte lacustrine diatomite deposit at Foulden Maar, Otago indicating the presence of Akaniaceae in southern New Zealand during the early Miocene. The flowers, although slightly smaller than the sole modern Australian species, A. bidwillii, contain pollen grains that are very like that taxon. The pollen also resembles that of the monospecific sister genus Bretschneidera from Southeast Asia and India, although that taxon has flowers with very different morphology from this genus. METHODS: The floral morphology of the fossil and in situ pollen grains were compared with flowers and pollen grains from extant species of Akaniaceae and related taxa. KEY RESULTS: The fossil inflorescence and associated pollen are referred to a new, extinct species of Akania: Akania gibsonorum. The floral structures and pollen resemble those of the modern Australian Akania species. CONCLUSIONS: The discovery of fossil flowers of Akania in an early Miocene lake deposit in New Zealand, coupled with earlier recognition of Akaniaceae leaves from the Paleocene epoch and wood from the Miocene epoch in South America suggests that the genus was once widespread in former Gondwana landmasses. The extinction of Akaniaceae in New Zealand and South America, and its present relictual distribution in eastern Australia, is most likely related to post-Miocene climatic cooling.

Hedycarya macrofossils and associated Planarpollenites pollen from the early Miocene of New Zealand.

PREMISE OF THE STUDY: The history of the basal angiosperm family Monimiaceae is based largely on fossil wood and leaf and floral fossils of uncertain affinity. Fossilized leaves with a well-preserved cuticle and Hedycarya-like flowers, including one with in situ pollen tetrads and fruits from an early Miocene lacustrine diatomite deposit in southern New Zealand implies a long record for Hedycarya in New Zealand. The flowers contain pollen grains that are very similar to those of the modern New Zealand species Hedycarya arborea and the Australian H. angustifolia but are considerably smaller. METHODS: We undertook comparative studies of the leaf, flower and fruit morphology of the newly discovered macrofossils and compared the in situ pollen grains from the flower with dispersed pollen grains from extant species. KEY RESULTS: The leaves are referred to a new, extinct species, Hedycarya pluvisilva Bannister, Conran, Mildenh. & D.E.Lee, (Monimiaceae), and associated with fossilized Hedycarya-like flowers that include in situ pollen and an infructescence of three drupes from the same site. Phylogenetic analysis placed the fossil into Hedycarya, sister to H. angustifolia in a clade with H. arborea and Levieria acuminata. A new name, Planarpollenites fragilis Mildenh., is proposed for dispersed fossil pollen tetrads at the site and those associated with the flower. CONCLUSIONS: The fossils are similar to those of modern Australian and New Zealand Hedycarya species, suggesting that the genus and related taxa have been significant components of the rainforests of Australia and the former Zealandian subcontinent for most of the Cenozoic.

Leaf fossils of Luzuriaga and a monocot flower with in situ pollen of Liliacidites contortus Mildenh. & Bannister sp. nov. (Alstroemeriaceae) from the Early Miocene.

PREMISE OF THE STUDY: The Foulden Maar lake sediments in Otago, South Island, New Zealand, date to the earliest Miocene and provide an important picture of the diversity of the Australasian biota, paleoecology, and climate at a time when New Zealand had a smaller land area than today. The diverse rainforest contains many taxa now restricted to Australia, New Caledonia, or South America. The presence of Luzuriaga-like fossils in these deposits is important for understanding Alstroemeriaceae evolution and the biogeography of genera shared between New Zealand and South America. METHODS: Leaves and a flower with in situ pollen that resemble extant Luzuriaga are described and placed phylogenetically. Geographic range information and a molecular clock model for the Alstroemeriaceae were used to investigate possible biogeographic scenarios and the influence of the new fossil on inferred divergence times. KEY RESULTS: Luzuriaga peterbannisteri Conran, Bannister, Mildenh., & D.E.Lee sp. nov. represents the first macrofossil record for Alstroemeriaceae. An associated Luzuriaga-like flower with in situ fossil pollen of Liliacidites contortus Mildenh. & Bannister sp. nov. is also described. The biogeographic analysis suggests that there have been several dispersal events across the Southern Ocean for the genus, with the fossil representing a now-extinct New Zealand lineage. CONCLUSIONS: Luzuriaga was present in Early Miocene New Zealand, indicating a long paleogeographic history for the genus, and L. peterbannisteri strengthens biogeographic connections between South America and Australasia during the Oligocene and earliest Miocene.

A fossil Fuchsia (Onagraceae) flower and an anther mass with in situ pollen from the early Miocene of New Zealand.

PREMISE OF THE STUDY: Fuchsia (Onagraceae) anthers, pollen, and an ornithophilous Fuchsia-like flower from an earliest Miocene lacustrine diatomite deposit at Foulden Maar, southern New Zealand confirm a long record for Fuchsia in New Zealand and probably an equally long history for its distinctive honeyeater pollination syndrome. The anthers contain in situ pollen of the fossil palynomorph previously assigned to Diporites aspis Pocknall et Mildenh. (Onagraceae: Fuchsia L.). • METHODS: We undertook comparative studies of the flower and anther morphology of the newly discovered macrofossils and compared the in situ pollen grains from the anthers with dispersed pollen grains from extant species. • KEY RESULTS: The anther mass is referred to a new, extinct species, Fuchsia antiqua D.E.Lee, Conran, Bannister, U.Kaulfuss & Mildenh. (Onagraceae), and is associated with a fossilized Fuchsia-like flower from the same small mining pit. Because Diporites van der Hammen is typified by a fungal sporomorph, the replacement name for D. aspis is Koninidites aspis (Pocknall & Mildenh.) Mildenh. gen. & comb. nov. Phylogenetic placement of the fossils agrees with a proximal position to either sect. Skinnera or sect. Procumbentes. These are the oldest macrofossils of Fuchsia globally. • CONCLUSIONS: The floral structures are remarkably similar to those of modern New Zealand Fuchsia. They suggest that the distinctive honeyeater bird-pollination syndrome/association seen in modern New Zealand was already established by the late Oligocene-earliest Miocene. The implications for the biogeography and paleoecology of Fuchsia in Australasia are discussed.

Poor quality vital anti-malarials in Africa - an urgent neglected public health priority.

BACKGROUND: Plasmodium falciparum malaria remains a major public health problem. A vital component of malaria control rests on the availability of good quality artemisinin-derivative based combination therapy (ACT) at the correct dose. However, there are increasing reports of poor quality anti-malarials in Africa. METHODS: Seven collections of artemisinin derivative monotherapies, ACT and halofantrine anti-malarials of suspicious quality were collected in 2002/10 in eleven African countries and in Asia en route to Africa. Packaging, chemical composition (high performance liquid chromatography, direct ionization mass spectrometry, X-ray diffractometry, stable isotope analysis) and botanical investigations were performed. RESULTS: Counterfeit artesunate containing chloroquine, counterfeit dihydroartemisinin (DHA) containing paracetamol (acetaminophen), counterfeit DHA-piperaquine containing sildenafil, counterfeit artemether-lumefantrine containing pyrimethamine, counterfeit halofantrine containing artemisinin, and substandard/counterfeit or degraded artesunate and artesunate+amodiaquine in eight countries are described. Pollen analysis was consistent with manufacture of counterfeits in eastern Asia. These data do not allow estimation of the frequency of poor quality anti-malarials in Africa. CONCLUSIONS: Criminals are producing diverse harmful anti-malarial counterfeits with important public health consequences. The presence of artesunate monotherapy, substandard and/or degraded and counterfeit medicines containing sub-therapeutic amounts of unexpected anti-malarials will engender drug resistance. With the threatening spread of artemisinin resistance to Africa, much greater investment is required to ensure the quality of ACTs and removal of artemisinin monotherapies. The International Health Regulations may need to be invoked to counter these serious public health problems.

Leaf fossils of the ancient Tasmanian relict Microcachrys (Podocarpaceae) from New Zealand.

PREMISE OF THE STUDY: Microcachrys tetragona (Podocarpaceae), endemic to the mountains of Tasmania, represents the only remaining taxon of one of the world's most ancient and widely distributed conifer lineages. Remarkably, however, despite its ∼150 Myr heritage, our understanding of the fossil history of this lineage is based almost entirely on the pollen record. Fossils of Microcachrys are especially important in light of recent molecular phylogenetic and dating evidence. This evidence dates the Microcachrys lineage to the Mesozoic and does not support the traditional placement of Microcachrys as sister to the southeastern Australian genus Pherosphaera. METHODS: We undertook comparative studies of the foliage architecture, cuticle, and paleoecology of newly discovered fossils from the Oligo-Miocene of New Zealand and M. tetragona and discussed the importance of Microcachrys in the context of Podocarpaceae phylogeny. KEY RESULTS: The fossils represent the oldest and first extra-Australian macrofossils of Microcachrys and are described as the new foliage species M. novae-zelandiae. These fossils confirm that the distinctive opposite decussate phyllotaxy of the genus is at least as old as the Oligo-Miocene and contribute to evidence that Microcachrys plants were sometimes important components of oligotrophic swampy habitats. CONCLUSIONS: Leaf fossils of Microcachrys closely comparable with the only extant species confirm that this lineage had a much wider past distribution. The fossil record and recent molecular phylogenetic studies, including that of Microcachrys, also serve to emphasize the important status of Tasmania as a refugium for seed plant lineages.

Poor quality drugs: grand challenges in high throughput detection, countrywide sampling, and forensics in developing countries.

Throughout history, poor quality medicines have been a persistent problem, with periodical crises in the supply of antimicrobials, such as fake cinchona bark in the 1600s and fake quinine in the 1800s. Regrettably, this problem seems to have grown in the last decade, especially afflicting unsuspecting patients and those seeking medicines via on-line pharmacies. Here we discuss some of the challenges related to the fight against poor quality drugs, and counterfeits in particular, with an emphasis on the analytical tools available, their relative performance, and the necessary workflows needed for distinguishing between genuine, substandard, degraded and counterfeit medicines.

A stratified random survey of the proportion of poor quality oral artesunate sold at medicine outlets in the Lao PDR - implications for therapeutic failure and drug resistance.

BACKGROUND: Counterfeit oral artesunate has been a major public health problem in mainland SE Asia, impeding malaria control. A countrywide stratified random survey was performed to determine the availability and quality of oral artesunate in pharmacies and outlets (shops selling medicines) in the Lao PDR (Laos). METHODS: In 2003, 'mystery' shoppers were asked to buy artesunate tablets from 180 outlets in 12 of the 18 Lao provinces. Outlets were selected using stratified random sampling by investigators not involved in sampling. Samples were analysed for packaging characteristics, by the Fast Red Dye test, high-performance liquid chromatography (HPLC), mass spectrometry (MS), X-ray diffractometry and pollen analysis. RESULTS: Of 180 outlets sampled, 25 (13.9%) sold oral artesunate. Outlets selling artesunate were more commonly found in the more malarious southern Laos. Of the 25 outlets, 22 (88%; 95%CI 68-97%) sold counterfeit artesunate, as defined by packaging and chemistry. No artesunate was detected in the counterfeits by any of the chemical analysis techniques and analysis of the packaging demonstrated seven different counterfeit types. There was complete agreement between the Fast Red dye test, HPLC and MS analysis. A wide variety of wrong active ingredients were found by MS. Of great concern, 4/27 (14.8%) fakes contained detectable amounts of artemisinin (0.26-115.7 mg/tablet). CONCLUSION: This random survey confirms results from previous convenience surveys that counterfeit artesunate is a severe public health problem. The presence of artemisinin in counterfeits may encourage malaria resistance to artemisinin derivatives. With increasing accessibility of artemisinin-derivative combination therapy (ACT) in Laos, the removal of artesunate monotherapy from pharmacies may be an effective intervention.

A collaborative epidemiological investigation into the criminal fake artesunate trade in South East Asia.

BACKGROUND: Since 1998 the serious public health problem in South East Asia of counterfeit artesunate, containing no or subtherapeutic amounts of the active antimalarial ingredient, has led to deaths from untreated malaria, reduced confidence in this vital drug, large economic losses for the legitimate manufacturers, and concerns that artemisinin resistance might be engendered. METHODS AND FINDINGS: With evidence of a deteriorating situation, a group of police, criminal analysts, chemists, palynologists, and health workers collaborated to determine the source of these counterfeits under the auspices of the International Criminal Police Organization (INTERPOL) and the Western Pacific World Health Organization Regional Office. A total of 391 samples of genuine and counterfeit artesunate collected in Vietnam (75), Cambodia (48), Lao PDR (115), Myanmar (Burma) (137) and the Thai/Myanmar border (16), were available for analysis. Sixteen different fake hologram types were identified. High-performance liquid chromatography and/or mass spectrometry confirmed that all specimens thought to be counterfeit (195/391, 49.9%) on the basis of packaging contained no or small quantities of artesunate (up to 12 mg per tablet as opposed to approximately 50 mg per genuine tablet). Chemical analysis demonstrated a wide diversity of wrong active ingredients, including banned pharmaceuticals, such as metamizole, and safrole, a carcinogen, and raw material for manufacture of methylenedioxymethamphetamine ('ecstasy'). Evidence from chemical, mineralogical, biological, and packaging analysis suggested that at least some of the counterfeits were manufactured in southeast People's Republic of China. This evidence prompted the Chinese Government to act quickly against the criminal traders with arrests and seizures. CONCLUSIONS: An international multi-disciplinary group obtained evidence that some of the counterfeit artesunate was manufactured in China, and this prompted a criminal investigation. International cross-disciplinary collaborations may be appropriate in the investigation of other serious counterfeit medicine public health problems elsewhere, but strengthening of international collaborations and forensic and drug regulatory authority capacity will be required.

Miocene Fossils Reveal Ancient Roots for New Zealand's Endemic Mystacina (Chiroptera) and Its Rainforest Habitat.

The New Zealand endemic bat family Mystacinidae comprises just two Recent species referred to a single genus, Mystacina. The family was once more diverse and widespread, with an additional six extinct taxa recorded from Australia and New Zealand. Here, a new mystacinid is described from the early Miocene (19-16 Ma) St Bathans Fauna of Central Otago, South Island, New Zealand. It is the first pre-Pleistocene record of the modern genus and it extends the evolutionary history of Mystacina back at least 16 million years. Extant Mystacina species occupy old-growth rainforest and are semi-terrestrial with an exceptionally broad omnivorous diet. The majority of the plants inhabited, pollinated, dispersed or eaten by modern Mystacina were well-established in southern New Zealand in the early Miocene, based on the fossil record from sites at or near where the bat fossils are found. Similarly, many of the arthropod prey of living Mystacina are recorded as fossils in the same area. Although none of the Miocene plant and arthropod species is extant, most are closely related to modern taxa, demonstrating potentially long-standing ecological associations with Mystacina.

An example of the use of forensic palynology in assessing an alibi.

A man was found shot in the back on Mount Holdsworth in the Tararua Ranges north of Wellington, the capital city of New Zealand. Police investigations pinpointed one individual who had been seen in the area, knew, and had the means and motive to kill the victim. His alibi was that an eyewitness was mistaken as he never had been in the area and the jacket he was reported to have been wearing had been purchased in The Netherlands and brought to Wellington, where it never had left the city. Furthermore the distinctive board shorts that he was reported to have been wearing had been purchased in a small coastal New Zealand town after the victim had been murdered. Pollen of Nothofagus menziesii, a mountain plant, on the clothing suggested that the alibi was untrue and that the clothing had been in mountains in the vicinity of Mount Holdsworth or a similar mountain scene where Nothofagus menziesii was growing.

Fossil Ericaceae from New Zealand: Deconstructing the use of fossil evidence in historical biogeography.

The Australasian Ericaceae epitomize many problems in understanding the biogeography of the southern hemisphere, especially the relative contributions of Gondwanan vicariance and dispersal. Late Cretaceous fossil pollen of the family suggests extreme antiquity of the group in Australasia, but recent phylogenetic evidence suggests much younger histories for most of the groups in that region. This paper documents two new species of latest Oligocene-Early Miocene macrofossils of Ericaceae from New Zealand. Cyathodophyllum novae-zelandiae G.J.Jord. & Bannister gen. and sp. nov. is the oldest record of the tribe Styphelieae, but is of a clade now extinct in New Zealand, possibly related to the Tasmanian genus Cyathodes. Richeaphyllum waimumuensis G.J.Jord. & Bannister sp. nov. is a member of Richeeae, but it is ambiguous as to whether it is a member of the impressive modern New Zealand radiation in Dracophyllum. These fossils emphasize the fact that at least some of the fossil pollen of Ericaceae may have been derived from extinct lineages and therefore should not be used as evidence for the antiquity of any modern New Zealand clade of Ericaceae. New fossils and/or detailed analysis of fossil and extant pollen may help resolve such uncertainty.