GOENCHE-JIVPASHAN-FOSSILS-OF-GOA-INDIA
Sunday, April 23, 2017
Friday, April 21, 2017
NOSTOC PRUNIFORME- THREE BILLION YEARS OLD LIVING FOSSIL SPECIES OF GOA
NOSTOC PRUNIFORME- THREE BILLION YEARS OLD LIVING FOSSIL OF GOA
World largest living organism by area occupied is a fungus
Armillaria found in Oregon, USA spread over
nine sq. kms. Goa’s largest living organism
by area could be the massive colony of the alga Nostoc pruniforme found at
Bambolim-Taleigao-Dona paula plateau spread over more than ten sq. kms.
This species can be declared as “official cyanobacterial species of Goa”. The millions
of Nostoc spherules are Goa’s own ,
least discussed “mare’s eggs”. Such sites like the one found at Harriman
Springs Resort, near Rocky Point, Oregon are preserved in USA for their ecological and
scientific value. But who cares for microbial heritage in Goa
when there are rich rewards to be reaped by harping on more familiar and
popular species?. It is amazing that in our own university campus such
sites are systematically being destroyed under the pretext of biodiversity
enrichment. We lost several locations where my students used to do ecological
studies on Nostoc spherules. Here no biological impact assessment is ever conducted when
earth moving equipments move in to blindly destroy entire habitats and
communities. The Nostoc empire on this plateau could also be one of the oldest
coastal prokaryotic colony. It is an ancient photosynthetic prokaryote, a
cyanobacteria which has the capacity to fix atmospheric nitrogen. Such a useful
natural species is completely neglected by landscape managers, agriculturists, photobiologists
and biotechnologists due to their ignorance and
indifference. The main monsoon vegetation on this lateritic plateau and similar coastal plateaus in Goa are the grasses. This property had compelled communidades of Goa to reserve such areas as “gavthans’ or community pastures. A rare species of Gondwana wild rices is also found on such plateaus. How these grasses can grow so fast and dense in just three months without any external nutrient subsidies in a nutrient poor soil?. I found the answer by closely looking at the root zones of the grasses. It is the processing of dry crust of dead Nostoc biomass which returns nitrogen rich nutrients to soil which are taken up by the roots of grasses. There are large patches of grass roots entwined by the Nostoc colonies. During the dry months if one pulls out a patch of these dry grass roots then a large dry Nostoc mat gets easily separated exposing the surface of the laterite. The plateau experiences a drying and wetting cycle with a five months wet spell and seven months dry spell. Nostoc colonization is maximum between June to middle of August. Once intensity and duration of rains decrease Nostoc colonies also become dry and almost unrecognizable. Like desert varnish the dry crust of these colonies create continuous greenish black carpets which outwardly look as cakes of cowdung but once intensive showers take place and sunshine is cut off these crustose mats suddenly revive and the action of movement of rainwater over the carpets creates something not found documented by anyone. Yellowish green to dark green, semi transparent cerebroid gelationous or jelly like spherules appear. These are often found in groups of 10 to hundreds. It is a joy to stand on the wind swept plateau with the rain beating on the umbrella and the rainwater washing your feet. And right in front of you is the magic of nature-a phenomenon totally neglected by all those blinded by only precision laboratory science. The Nostoc spherules get dislodged with flowing turbulent sheets of water and begin rolling on the plateau, while getting entangled by the grass roots, natural depressions and cavities and crevices in the spongelike lateritic rock surface. I have seen translocation of some spherules for a distance of several metres before their final entanglement. Generally the rolling speed is high on an incline or gradient and slow on a flat surface. The force of the current also disperses these spherules over large distances. So colonies of same species get dispersed year after year and then come to establish a giant clonal colony. The Nostoc pruniforme spherules collected and examined by my students showed uniquely buoyant property when dropped in water indicating their low density and probable presence of air cavities in the gelatinous matrix. The gelatinous nature also showed presence of an unidentified compound with exceptional swelling properties- similar to Jalshakti water absorbing synthetic polymer foam produced by National Chemical laboratory, Pune. We did not try using powdered dry and old biomass of Nostoc to check whether it can be revived as spherules after absorbing water. The unique wetting, swelling, expanding and then drying, contracting, collapsing feature of spherule forming ecotype, genotype and biotype of Nostoc pruniforme makes it a special case in algal biotechnology. This form is sensitive
to humidity, temperature and light and prefers high to very high humidity, moderate temperature and only diffused light. Direct sunlight causes the spherules to shrink. This species is Goa’s natural factory of nitrogenous biofertilizers. If its’ wild habitat is conserved and studied scientifically then intelligent scientific clues can be obtained to control its’ production. The relative ease of drying the spherules after water loss makes it an excellent candidate for mass propagation and biofertilizer formulation. Nostoc spherules could be easily mistaken as heaps of herbivores dung. Their colour merges with the ground vegetation. During the dry spell the spherules simply vanish and one finds a thin greenish mat on their original location. Such morphological mode switching from two dimensional stationary mats to three dimensional rolling, dynamic spherules and vice versa remains relatively under investigated in the area of phycology- the science of study of algae. This property also makes the species another excellent model in developmental biology and molecular morphogenesis. A time lapse series of photographs of the filamentous mat transforming into rolling spherules would be so interesting that it would open a new line of research in ecological genetics. The “mare’s eggs’ of Goa, is a living fossil, a primary producer, with high photosynthetic and Nitrogen fixing capacity and deserves its’ due place among the list of important species to be protected. Compared to known species there is more danger of extinction of relatively less known or unknown species like Nostoc pruniforme which may simply vanish after land use change, habitat modification and ecological simplification. The spherules have evolved in tune with the rocky plateau. The rare spherule forming Nostoc pruniforme , the unsung cyanobacterial species of Goa is as important as tigers and turtles, frogs and fish. But who cares?. Even the agriculture department has not heard of this species.
indifference. The main monsoon vegetation on this lateritic plateau and similar coastal plateaus in Goa are the grasses. This property had compelled communidades of Goa to reserve such areas as “gavthans’ or community pastures. A rare species of Gondwana wild rices is also found on such plateaus. How these grasses can grow so fast and dense in just three months without any external nutrient subsidies in a nutrient poor soil?. I found the answer by closely looking at the root zones of the grasses. It is the processing of dry crust of dead Nostoc biomass which returns nitrogen rich nutrients to soil which are taken up by the roots of grasses. There are large patches of grass roots entwined by the Nostoc colonies. During the dry months if one pulls out a patch of these dry grass roots then a large dry Nostoc mat gets easily separated exposing the surface of the laterite. The plateau experiences a drying and wetting cycle with a five months wet spell and seven months dry spell. Nostoc colonization is maximum between June to middle of August. Once intensity and duration of rains decrease Nostoc colonies also become dry and almost unrecognizable. Like desert varnish the dry crust of these colonies create continuous greenish black carpets which outwardly look as cakes of cowdung but once intensive showers take place and sunshine is cut off these crustose mats suddenly revive and the action of movement of rainwater over the carpets creates something not found documented by anyone. Yellowish green to dark green, semi transparent cerebroid gelationous or jelly like spherules appear. These are often found in groups of 10 to hundreds. It is a joy to stand on the wind swept plateau with the rain beating on the umbrella and the rainwater washing your feet. And right in front of you is the magic of nature-a phenomenon totally neglected by all those blinded by only precision laboratory science. The Nostoc spherules get dislodged with flowing turbulent sheets of water and begin rolling on the plateau, while getting entangled by the grass roots, natural depressions and cavities and crevices in the spongelike lateritic rock surface. I have seen translocation of some spherules for a distance of several metres before their final entanglement. Generally the rolling speed is high on an incline or gradient and slow on a flat surface. The force of the current also disperses these spherules over large distances. So colonies of same species get dispersed year after year and then come to establish a giant clonal colony. The Nostoc pruniforme spherules collected and examined by my students showed uniquely buoyant property when dropped in water indicating their low density and probable presence of air cavities in the gelatinous matrix. The gelatinous nature also showed presence of an unidentified compound with exceptional swelling properties- similar to Jalshakti water absorbing synthetic polymer foam produced by National Chemical laboratory, Pune. We did not try using powdered dry and old biomass of Nostoc to check whether it can be revived as spherules after absorbing water. The unique wetting, swelling, expanding and then drying, contracting, collapsing feature of spherule forming ecotype, genotype and biotype of Nostoc pruniforme makes it a special case in algal biotechnology. This form is sensitive
to humidity, temperature and light and prefers high to very high humidity, moderate temperature and only diffused light. Direct sunlight causes the spherules to shrink. This species is Goa’s natural factory of nitrogenous biofertilizers. If its’ wild habitat is conserved and studied scientifically then intelligent scientific clues can be obtained to control its’ production. The relative ease of drying the spherules after water loss makes it an excellent candidate for mass propagation and biofertilizer formulation. Nostoc spherules could be easily mistaken as heaps of herbivores dung. Their colour merges with the ground vegetation. During the dry spell the spherules simply vanish and one finds a thin greenish mat on their original location. Such morphological mode switching from two dimensional stationary mats to three dimensional rolling, dynamic spherules and vice versa remains relatively under investigated in the area of phycology- the science of study of algae. This property also makes the species another excellent model in developmental biology and molecular morphogenesis. A time lapse series of photographs of the filamentous mat transforming into rolling spherules would be so interesting that it would open a new line of research in ecological genetics. The “mare’s eggs’ of Goa, is a living fossil, a primary producer, with high photosynthetic and Nitrogen fixing capacity and deserves its’ due place among the list of important species to be protected. Compared to known species there is more danger of extinction of relatively less known or unknown species like Nostoc pruniforme which may simply vanish after land use change, habitat modification and ecological simplification. The spherules have evolved in tune with the rocky plateau. The rare spherule forming Nostoc pruniforme , the unsung cyanobacterial species of Goa is as important as tigers and turtles, frogs and fish. But who cares?. Even the agriculture department has not heard of this species.
ON THE TRAIL OF GOA'S PLANT MEGAFOSSILS
ON THE TRAIL OF GOA'S PLANT MEGAFOSSILS
I have a piece of fossilized or silicized wood which was found during 1966-67 while building an approach road on Borim’s Siddanath hill. How fossilized wood can be found at such an altitude?. Except NIO’s Dr. Rajiv Nigam no scientist has paid any consistent attention to research on Goa’s marine fossils which are still being mined for making mortar. The criminal neglect of fossils in Goa shows how little we care for challenging research. We should feel ashamed that after 1876 neither the government -Portuguese or Indian, nor the scientists and academicians followed this interesting lead which would have put Goa on world map of plant megafossils and could have boosted tourism in western ghats. Isabel, Lady Burton (1831 -1896) — née Isabel Arundell — was the wife and partner of explorer, adventurer, and writer Sir Richard Francis Burton (1821-1890). Sir Richard Burton had visited Goa and had written an account-Goa and the blue mountains in 1851. But the great adventurer missed what his wife later found. Lady Burton’s visit took place somewhere in 1875-6. This is how she described her experience of visiting a site of petrified forest in Goa -“We made two boat expeditions together -- one to see a coffee plantation, in which is a petrified forest. Each expedition occupied two or three days. We embarked for the first in a filthy boat, full of unmentionable vermin, and started down the river in the evening, with storms of thunder and lightning and wind precluding the monsoon. On arrival we toiled up two miles of steep, rocky paths through cocoa groves. At the bottom of the hill was a little rivulet, and pieces of petrified wood were sticking to the bank. As we ascended the hill again we found the petrification scattered all over the ground; they were composed chiefly of palms and pines; and most interesting they were. “ (Source: Burton, Lady Isabel (1831-1896).Wilkins, W. H. (1860-1905). The Romance of Isabel Lady Burton Volume II, The Story of Her Life, Told In Part by Herself and In Part by W. H. Wilkins. Chapter 22). After 136 years I was probably following her trail blindly on an unusually warm November afternoon. This is a short report on my discovery of a new and exciting location of petrified forest-meaning plant/tree megafossils in Goa. This is not the same location reported by some fossil hunters in May 2012. They were actually confirming another 1876 report. I have inspected that location also and obtained samples for studies. Goa is land of micro, macro and megafossils. Marine fossils are plundered from Camurlim, Tuem, Agapur for making lime. Marine fossils are also located near Mapusa river. Sancoale or Shankhavali derives its’ name from fossilized marine conches and windowpane oyster shells found in the soil there. There is belt of marine fossils from Cortalim to Chikhalim and Bambolim to Siridao on both the banks of Marmagoa bay. Since the trained scientists, except NIO’s Dr. Nigam and his student have not cared to study these and such sites I was compelled to undertake the field and lab investigations. Work done by Dr. Nigam’s students on marine fossils of Konkan showed relict presence of T. squamosa. They explained it by the fact that it resided in this area and got encrusted on foraminiferal tests when the sea level was lower at ca. 10,000 yrs BP, thus indicating that at ca. 10,000 yrs BP, the Konkan shoreline was 80 m lower than that at present. No historian in Goa has understood the implications of such lower sea level for human migrations. The finding was published in January 2013 issue of GOA TODAY because there is an unhealthy and unethical competition to take false credit. I experienced it when rock art gallery was discovered in May 1993 at Usgali/Panasaimal,Kolamb, Rivona (Goa Today, June 1993). Hoards of fossil hunters may descend in Goa after publication of this article. Goa has violent tectonic past which resulted in rocks being folded and faulted, rivers changing their courses and entire forests becoming petrified. The legend of Parashurama abstracted these events. Portuguese suspected it. Lopes Mendes mentioned it. German geologist Oertel believed it. Anant Dhume was first historian to point to marine fossils in Sattari. He found a marine conch at Thane, Sattari and hinted at fossils at Ivrem khurd and budruk villages. He had even donated a petrified root specimen to Goa museum where it is not now traceable. During 1966-8, a road construction was undertaken on Bori hill. Mr. Percival Noronha, the then director of tourism, found fossilized tree trunks and petrified logs by the roadside and managed to retrieve a sample which he later gifted me. It was identified by Goa University geologists as silicized wood. However, no expert on geology of Goa could explain to me how plant megafossils could be found at an altitude of several hundred metres from present sea level. Then in upper reaches of Mhadei river basin I could detect rare fossilized stromatolites, the most ancient photosynthetic marine bacteria on earth, in photographs taken by Mr. Prakash Poreinkar during a three day trek to the source of river. This was first report of such ancient microfossils (http://www.navhindtimes.in/panorama/neglected-stromatolite-fossils-goa) anywhere on west coast and western ghats of India. During my field trip in November 2012 for specific investigations of petrified forests reported by British geological adventurers in Sattari (Marchesetti, C. D. On a Pre-historic Monument of the Western Coast of India. Journ. Bombay Branch B. As. Soc. vol. xii. pp. 215-218. 8th April 1876) guided by Prakash Porienkar, I discovered on the banks of Mhadei river and in the semi dry channel as well at a location called – JAGHDHA ( location name changed) thousands of sq. mts of petrified forest comprising huge horizontally piled up petrified tress possibly monocots. The local rock formation is Vageri meta-greywacke/metabasalt in a banded Iron formation zone. The dating is difficult and would be shortly done by Birbal Sahani institute experts. B massive and deeply layered , richly textured, very hard but brittle plant megafossils are different than 20 million years old petrified trees found at the National Fossil Wood Park in Thiruvakkarai, located about 35 km from Puducherry. These belong to what is called the ‘Mio-Pilocene' age. GSI experts say that the these fossil woods was the result of a huge flood that occurred millions of years ago and destroyed large parts of forests in the adjoining area. The flood story for Goa is unknown. However going by the hard metabasaltic nature the the JAGHDHA , Sattari fossils may be as old as fossils of Akal Fossil Wood Park, Jaisalmer . These petrified wood carries signature of the luxuriant forests in a warm and humid climate, bordering the sea some 180 m.y. ago. The fossils are of petrophyllum, ptyllophyllum, equisetitis species and dicotyledonous wood and gastropod shells of Lower Jurassic period. The JHADHA Sattari megafossils also differ from the petrified specimens in National Fossil Wood Park in Sattanur, Perambalur District which contains large trunks of petrified trees of Upper Cretaceous age (100 m.y). The trees belong to conifers (non-flowering) that dominated the land vegetation during the period. The fossilised tree trunk at Sattanur measures over 18 m in length. The JHAGHDA petrified trees are horizontally piled up, and extend several metres in the river channel. The petrification event may be associated with the violent tectonic event of change in the course of Mhadei river. Experts have opined that this area was a geological high ( at least above 800 metres) when volcanism in Deccan, 63-66 million years ago formed the flood basaltic plateau. Fossil specimens which I have examined in laboratory clearly showed intact petrified vascular bundles, a lot of pyrite ( Iron sulphide) , black sulphides of Gold, silica grains and auriferous quartz. On basis of ecological, hydrological, local geological evidence and mineralogical basis, I wish to tentatively date these megafossils to 100-180 my may be more but dating is tricky and expert confirmation is awaited. My attempts since November 24 to contact the Chief Minister and arrange his visit to this sensational fossil site failed. I would try again. Only he can issue proper orders. The easily accessible and unprotected area needs surveillance otherwise within no time it would be plundered. A more detailed report would be published in this magazine later. The local village panchayat, the forest, the water resources department and collector, north Goa should take steps to protect this site immediately. It is actually a continuous belt of massive petrified forest extending from Bhironda to Khotodem and exposed only by the channel of river of Mhadei. The part which is buried is difficult to trace but may be exposed in trial trenches. We need to be grateful to Lady Burton for awakening us. Here is the nucleus for Goa’s new geological park.
THE NEGLECTED STROMATOLITIC FOSSILS OF GOA
The neglected Stromatolitic fossils of Goa
What are “stromatolites” ?. These are internally-laminated,
macroscopic sedimentary structures, commonly of biological origin which form
the dominant part of Earth’s early fossil record and so provide a potentially
important source of information about early life. This article reports the new
discovery of Archean (more than 2500 million years old) stromatolites-vast
fossilized algal mats on border of Goa, inside
Mahadayi river basin among metasedimentary rocks. Time is running out for
fossil researchers because the strong currents of Mahadayi have eroded the
stromatolites. These mats are not the routine major or minor folds found in
metasedimentary rocks. There are stromatolitic carbonates as well. A group of
trekkers had visited the source of Mahadayi river. In the bed of the river and
on the banks they came across strange rocks. The students of Geology amongst
them could not identify these strange formations. One of the leader of the
trekkers, Prakash Parienkar who teaches at Konkani department of Goa University
showed me the photographs. One of these is published alongwith this article.
After minute examination of the images and based on geology and
lithostratigraphy of the Mahadayi basin which is full of detrital Archean
metasediments and after consulting other published sources on stromatolitic
deposits in India and outside, I arrived at the conclusion that upper Mahadayi
basin is a vast, unexplored, neglected treasure trove for archean fossil
hunters with stromatolite fossils being relatively abundant. But the currents
are eroding these fossils. It is virtually a Archean paleontological museum.
Only fools would ignore its’ monumental scientific, academic , intellectual
potential. The Mahadayi river basin is part of Western Dharwar Craton
(WDC) of Meso-archaean age. WDC shows greenstone belts, tonalite-trondhjemite
gneisses, high-Mg basalts, komatites with metavolcanics and meta-sedimentary
rocks. Among these mafic and ultramafic volcanic rocks, arenites, phylites,
polymictic and oligomictic conglomerates, greywackes, banded iron formations
and carbonates are dominant. What’s practical importance of discovery of
stromatolite fossils in Goa?. My attention was
drawn to Geologist Hofmann’s 1973 paper on stromatolites from which I take
liberty to quote the relevant part- “ Stromatolites are remarkable structures.
They are so primitive and simple, and yet so complex and versatile. Not so long
ago still regarded by some as curious objects, these laminated biosedimentary
structures are now intensively studied. They provide information whose
importance spans more than 2.9 billion years of time and three of the major
fields of science: geology, biology and astronomy. Within geology, they have
contributed information of use in sedimentology, paleontology, stratigraphy,
paleogeography, geophysics, and even tectonics. They have been called upon: (1)
to identify places of past biotic activity; (2) to interpret ancient
depositional environments and paleosalinities; (3) to make correlations and age
determinations; (4) to place limits of the time of origin of cyanophytes, oxygen-producing
photosynthesis, filamentous habit of algae, and integrated biological
communities; (5) to determine paleocurrent directions; (6) to map ancient
shorelines; (7) to measure past tidal ranges; (8) to time the closest approach
or the capture of our moon; (9) to determine ancient astronomical rhythms such
as the length of the synodic mont h and the number of days per year; (10) to
determine rates of sediment accumulation; (11) to determine paleolatitudes;
(12) to prospect for base metal deposits; (13) to prospect for microfossils;
(14) to make top-determinations in deformed sequences; (15) to serve as tourist
attractions.” A huge amount of work has been done after Hoffman’s well
delivered paper at the 46th Annual Meeting of the Society of Economic Paleontologists
and Mineralogists, April 18, 1972, Denver,
Colorado. Extraordinary claims
need extraordinary evidence. Late Anant Dhume who wrote “cultural history of Goa” as surveyor had toured the whole state in his field
based career of more than half a century. He could read geomorphology of Goa like a book. While editing his book I gained a lot of
new knowledge about paleontology of Goa. He
had mentioned about discovery of a marine conch at Rive, Thana, Sattari at a depth of half metre. I
had asked him in 1984- “how is it possible to get marine fossils in
Sattari, that too at an altitude of few hundred metres and more than 50 kms.
from the sea?”. He attributed it to paleotectonic activity. The detrital
metasediment in Mahadayi basin points to the possibility of violent tectonic
activity before the formation of Deccan flood
basalt and orogeny of western ghats. Before publication of this article people
would not have believed that the upper basin of Mahadayi
river in Sattari was once ( at least 2.5-3 billion years ago) below the sea or
was a shallow paleoseashore/ paleointertidal zone or may be even a paleoreef.
And this sea or seashore was teeming with life as shown by the extraordinarly
rich stromatolitic fossil rocks. Why stromatolite fossils were not previously
reported?. Firstly there is poor emphasis on paleontology in education and
research. The biogenic factors are often forgotten. Then these rocks are not
located in a single place. Most of the time the currents mask or
hide them. The exposed ones get mixed and easily camouflaged within the
surrounding metasedimentary rocks. Not every folded rock is a stromatolite. A
discovery can be made only if focussed, consistent, rational sampling efforts
are done. This is a region of upper Mahadayi basin which no scientist from NIO,
Goa University, GSI or any other educational
or research institutions has ever dared to enter. The terrain and the route is
treacherous. The route is known only to the local people. Visit is possible
only during late winter. According to experts, Stromatolites are shaped by a
complex interaction of physical, chemical, and biological processes, and
identifying unambiguous signatures of life from the preserved morphology of the
structures can be extremely difficult. There is also need to identify the
activities of benthic microbial communities. But such attempts face problems
due to the process of diagenetic alteration, particularly recrystallization: a
diagenetic process that commonly affects the chemical (precipitated) sediments
with which stromatolites are often associated. Goa
is teeming with such macro and microfossils. But palaeontology of Goa is a totally neglected subject.
Fossilized Microbial Forms in Baltic and Goan Amber- A Comparative Pioneer Study
Fossilized Microbial Forms in Baltic and
Goan Amber- A Comparative Pioneer Study
Dabolkar Sujata* and Kamat Nandkumar
Department of Botany, Goa University
, Taleigao, Goa, 403206 , India
Corresponding Author Email : nandkamat@gmail.com
*Presenting Authors Email :
sujatadabolkar@gmail.com
ABSTRACT
This
paper is based on surveys, exploration
and standardization of techniques to recover rare amber samples from sands
in Goa and identify specimens embeded with distinct microbial fossils based on
studies on reference samples of imported Baltic Amber. We developed techniques
to locate, detect and identify amber samples in local sands. In this pioneer
study we report presumptive microbial forms such as Actinobacteria and fungi in
amber samples of Goa throwing light on microbial paleobiodiversity. Baltic
amber (Succinate) is fossilized resin
belonging to the Eocene period (44-49 million years old) derived from the Araucariaceae and Leguminoseae family of trees containing up to 8% of Succinic acid
and compounds such as terpenoids and phenolic derivatives. Sooty moulds in the
amber fossils have been studied (Schmidt et al., 2013). Samples of imported Baltic
amber were validated, studied and used as reference for identification and
characterization of amber found in rivers sands of Goa. FTIR Spectroscopic tests
diagnostic of presence of Succinate proved that both samples meet the criteria
as plant derived Succinate containing products. Fossil fungi in Baltic amber
were observed, and compared with similar forms in rare amber fragments of Goa.
These samples were subjected to butanol treatment and microscopic examination.
Actinobacterial and fungal forms embedded in local amber were compared with similar
forms found in imported Baltic Amber and those published in literature. Detection
of fossilized Actinobacterial and fungal forms has shown us the potential for further
studies for comprehensive collection and microscopic examination of such paleomicrobial
forms in rare local amber samples.
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