Visible by Nadia Rayward

VISIBLE Animals in Captivity Racehorses

Landscapes

Water and City Life

NEW ! Small Worlds In Vitro Fertilization

Urban Wildlife

Turtles in Madrid

Birds

Carmel by the Sea Nadia Abou-Hinin Rayward MSc Biological Photography and Imaging Professional Techniques CD14204 Student ID 4200866 1

Contents Editorial

Birds Carmel by the Sea

Landscapes Water and City Life

Animals in Captivity Racehorses

Small Worlds In Vitro Fertilization

Urban Wildlife Turtles in Madrid

Bibliography, Maps & Technical Data

Visible 3

One of the multiple types of orchids found in the cloud forest of Mindo, Ecuador.

Visible

Editorial T

he May edition of VISIBLE is ready to go to press. The Editorial Board is especially excited about this new edition. The Board has been debating for the past months whether or not to add a new feature to the journal. There was much discussion amongst the members of the Board if VISIBLE wanted to move forward with a new idea. In general the Board believed that every year we should introduce a change, it has been 14 months since we introduced Landscapes. It was voted that it was time to renew while following VISIBLE’s dedication to biological photography with an emphasis on wildlife. Many ideas were submitted for approval and it was a tedious choice. There were ideas such as Artic Photography; Exotic Plants just to name a few. The most interesting proposal had one real problem for some of the members of the Board. This problem eradicated in the fact that the feature was not true to the journal’s name. The new feature section that was being proposed was invisible: it had to do with microscopy! Some members of the Board argued that microscopy was not visible, but other members debated that what about camera lenses? At the end, consensus was achieved and the May issue of VISIBLE introduces a new monthly feature article on microscopy. We have called this feature “Small Worlds”. After the Board saw the amazing photographs published this month, there was no doubt in anybody’s mind that the correct decision had been taken. The article to inaugurate this new section is about what goes on in an IVF lab. The article reviews human reproduction in general and gives a basic outline on techniques used in assisted human reproduction. We are sure you are going to enjoy the stunning photography: from gametes to 5-day-old embryos called blastocysts. We can vicariously experience the miracle of life from its very beginning. We truly hope that you will be as keen as we are on the new monthly feature. As always we would appreciate your feedback: feedback@visible.com. We wish you an enjoyable read of this month’s journal. As always we are open to your point of view to help us make VISIBLE the most beautiful and informative monthly biological journal on earth.

Happy month of May!

Editorial Board of VISIBLE.

A selection of birds seen at 6

Birds at sunset, Point Lobos, Carmel by the Sea.

Carmel by the Sea

armel-by-the-Sea or just Carmel is a city in Monterey County in Central California. Carmel is located on the Pacific Ocean, about 120 miles south of San Francisco. Carmel is known for its scenery, its wildlife, its artistic population and eccentric laws. One of these laws in the Municipal Code is that no one can wear heels over 2 inches. The law dates from 1910 and was created due to the uneven pavement in the city. Carmel is also known as the most dog-friendly city in the US because dogs are allowed just about everywhere in Carmel: they are permitted in most hotels and restaurants and some of these restaurants even have doggie menus (1). Due to the diversity of habitats, Monterey County is a wonderful place for birding. There are 427 species listed in the county (2). It has beautiful white-sanded beaches with tall close-coned conifers especially the Monterey cypress and Monterey pine. There are isolated rocks offshore, which provide adequate space for nesting. 8

A famous part of the Monterey Peninsula is the 17-mile drive, which is home to the Pebble Beach Golf Course and a gated community. One of the entrances to the 17-Mile Drive is from Carmel. The road runs in part adjacent to the coastline and up and down the hills, which adjudicates many locations spectacular viewpoints. The two most famous scenes on the 17-Mile Drive are Bird Rock and the Lone Cypress- the official symbol of Pebble Beach. Another natural attraction just south of Carmel is Point Lobos State Natural Reserve. In the ocean around Point Lobos there are large kelp forests. These kelp forests of marine algae provide food and shelter for a broad scope of marine life from shrimp and fish to sea otters, sea lions and a variety of birds. The Reserve has a rocky coastline dotted with sandy coves and small islands which attracts migratory as well as nonmigratory birds (3). The first two birds described are less common than the crow and the seagull but they all make up the beautiful combination of wildlife and scenery that makes Monterey County what it is.

Map pag.8: Carmel-by-the-Sea location in relation with the US. All pictures in this article wew taken in this location. Map pag.9: 17-Mile-Drive location. Picture pag.9: Picture taken in the 17-Mile-Drive. View of the coast in Carmel-by-the-Sea, California.

Brandt’s Cormorant Brandt’s Cormorant or Phalacrocorax penicillatus, is named in honor of the German naturalist Johann Friedrich von Brandt (4) who first described the bird at the beginning of the 19th century. Its specific name Pencillatus makes reference to the white plumes that appear on its neck and back. These plumes only appear during the early breeding season. In Latin, Pencillatus means a pencil of hairs or painter’s brush. They live on the Pacific coast from Alaska to the Gulf of California in North America. The Brandt’s Cormorants, which live around Carmel, are not migratory birds. The ones that live further north do migrate south in winter. These marine and terrestrial birds nest mostly on flat ground or on slopes (5). Other times they nest on sandy beaches or on rocks. 10

Brandt’s Cormorants feed on small fish, mostly rockfish in California. They can dive up to 150 feet below the surface. They use their highly developed feet for propulsion. These birds are usually silent but when they are in a nesting colony, sometimes they make give deep, low grunts and croaks.They are abundant in number although the population seems to be decreasing but Brandt’s Cormorant is not a threatened species. Laying occurs from March to July. They live easily with other species of birds. Picture pag. 10: Phalacroconax penicillatus. Picture pag.11 top: Larus occidentalis (juvenile). Picture pag.11 bottom: Larus occidentalis. All of these pictures were taken at the coast of the 17-Mile-Drive.

Western Gull The Western Gull or Larus occidentalis is another conspicuous and common species that lives along the Monterey coast. These are the classic “seagulls” (5). These seagulls have white heads and grey wings. They measure between 22-27 inches. These seagulls nest very near to the sea, on the coast or on the rock islands just offshore. They nest singly or in colonies. The pair builds a nest lined with vegetation. Three eggs are usually incubated and hatch 29-32 days later. The young are fed by their progenitors’ regurgitation. They feed on anything edible they find and even use landfills to find food. They also can fish on the surface of the sea and find krill, jellyfish and squid. The chicks must stay within the territories of their progenitors. If they wander out of their territory, the pair whose territory they have strayed into may even kill them.They tend to make lots of noise with a typical kuk kuk kuk protecting their young or when feeding (5).

American Crow The American Crow or Corvus corax is a very common bird in North America. It is a very visible and adaptable species throughout the lowlands of California and all of North America (4). Interestingly enough it is very susceptible to the to the West Nile Virus and is used as a bioindicator of the disease. Because of the introduction of the disease in the US in 1999 it is estimated that the American Crow population has decreased in by 40% (6). Even though there has been this tremendous decline the crow is not a species of concern. It has an all-black body, bill and legs and measures 37 inches (5) and their tail makes up about 40% of its size. Its feathers are iridescent black. A fascinating fact is that the American Crow has a distinct loud and short caaw-caaw-caaw sound and moves its head up and down when it makes noise, but it are also capable of imitating sounds other animals make especially other birds. These birds are inhabitants of big cities, woodlands and by the sea in Monterey County.

They are omnivorous and eat invertebrates of all types. They feed on carrion, other bird’s eggs, berries, seeds and human food waste. They nest in a wide diversity of trees and locations: steep cliffs, ledges and bluffs and rarely near the ground. Their nests are made of sticks and branches and then the birds line the nests with soft plant materials and fur (5).

population seems to be decreasing worldwide it is not a threatened species on the IUCN Redlist (4). They like mudflats, slat-water marshes and rocky shorelines (5). They feed by probing and pecking at the surface of mud or sand eating ground insects, mollusks, crustaceans and worms.

Red Knot

The birds in Monterey County are diverse and abundant. This is truly a bird watchers paradise. Not only are the birds spectacular but the coastline with splendid trees, sand coves, rocks and small rock islands enhance the natural beauty of this part of the world.

The Red Knot or Calidris canutus is a shorebird, which breeds in the Artic (5). It is a medium-sized bird (about 10 ½ inches with a wingspan of 18 ½ -21 inch wingspan) and is not a numerous species along the California coast. In winter its plumage is a basic grey. There is virtually no difference in plumage between sexes in winter. When they breed sex may be distinguished by plumage. It is usually silent but sometimes softly calls its name, knot. Although the Red Knot

Monterey County and its bird population

Picture pag.12: Covux corax. Picture pag 13: Caladris canutus. Both pictures were taken in the coast of Carmel-bythe-Sea.

Water & City Life

Sevilla and the Guadalquivir river, Spain.

eville, Miami Beach, San Francisco, Paris and London: what do they have in common? Water! Seville is on the Guadalquivir River, Paris on the Seine, London on the Thames and Miami Beach is on the Atlantic Ocean and San Francisco is on the San Francisco Bay.

Seville Seville is the fourth largest city in Spain. It is 80 km form the Atlantic Ocean and is a river port. It is the only river port in Spain. Guadalquivir means the big river in Arabic . The river plays an important part in the city. The cathedral, the Reales Alcรกzares Palace and the Barrio de Santa Cruz are all located on the east back. The Barrio de Santa Cruz is small disctrict of Seville made up by a labyrinth of little streets, bars, 16

restaurants and shops along that have been opened amid traditional Andalusian houses with tiled patios filled with ferns and geraniums. On the west bank, the neighborhood of Triana and Isla Mรกgica a theme park, are found. In the Gualdalquivir River there is an almost-island called Isla de la Cartuja. The famous bridge designed by controversial architect Calatrava, connects the almost-island to Seville proper. There are many restaurants on the riverbanks, which serve typical Andalusian food: fried fish, gazpacho, Spanish ham, cheeses and tortilla. It is what makes Seville, Seville. There are promenades right by the water where people stroll and relax in the sun, jog and read.

Even though Seville is not a big city, it does offer river cruises, which are not only relaxing but also informative, as many important buildings in Seville are along the Guadalquivir. An interesting example is the â&#x20AC;&#x153;Torre del Oroâ&#x20AC;? or Gold Tower, which is a dodecagonal military watchtower, built by the Almohad dynasty in 1120, to able the authorities to control the access to Seville by river. It is called the Gold Tower not because it was made up of gold but because it stored the gold that was brought from America. That is one of the legends. Another is because when it was first constructed it had shiny tiles. At some points of the river kayaking, rowing and canoeing are allowed. All and all Seville is proud to have the only river port city in Spain and to have such a beautiful river running through it. The river provides changing scenery and views of the city.

Picture pag.16: Areal shot Miami Beach . This picture was taken from a jet. The different bridges to get to mainland Miami can also be appreciated in this picture. Map pag.17: Location of Miami in relation with Florida and the Caribbean.

Miami Beach, even though it has been overdeveloped is still an awe-inspiring spot for artists and photographers.

Miami Beach Miami Beach is a beach city on the Atlantic Ocean in the state of Florida, USA. Actually Miami Beach is made on a series of islands- some are natural and others are man-made. The islands are between the Atlantic Ocean and the Biscayne Bay. For over 100 years Miami Beach has been the most important beach resort in the United States. Water sports like waterskiing, sailing, speed boating and kite surfing are very popular. People enjoy relaxing on the beaches splashed with colorful beach umbrellas. The city is filled with ethnic restaurants and boutiques but the main attraction is undoubtedly is the beach. South Beach is the surf capital of Miami Beach. Not only can you enjoy watching top surfers do their thing but the main road called Ocean Drive is full of beautiful suntanned people strolling, rollerbladers and some very interesting people to look at. While on the beach it is easy to join people who want to play volleyball or soccer. Another possibility for doing something by the water is taking a surf lesson. Little surf boutiques are plentiful on the beach. Kayaking, diving, fishing and snorkeling are also popular. Virtually anything that has to do with water or water sports can be found in Miami. The view of Miami Beach from the air is breathtaking. The different shades of blue and green of the water change constantly. In just a few hours the colors of the ocean can change from turquoise to dark green and then all of a sudden there are spots of kelly green and light blue. Tall buildings right on the beach show how over-developed this area is. Being such a beautiful venue it is really a pity to see such a natural wonder so overly populates. The sky also frequently changes. This is very true of all tropical climates. Rain comes and goes and leaves behind beautiful rainbows. There are sometimes clouds, especially in the rain season that give almost surreal lighting to the city. 17

San Francisco San Francisco is on the San Francisco Bay. Water from the Sacramento and Joaquin rivers and the Sierra Nevada mountains passes through the bay on its way to the Pacific Ocean. Five large islands are located in the bay. One of them is Alcatraz, which served as a prison until 1963. It is definitely one of the great tourist attractions of San Francisco. There are also lots of boats for tourists to take for sightseeing around the bay or going to destinations around San Francisco like Sausalito, a charming artistic enclave where many inhabitants live on houseboats. Also leaving from San Francisco Port are boats that serve dinner and go on a tour of the bay to see the lights of San Francisco. Another attraction on the Bay is the exceptional Golden Gate Bridge. You can walk, bike or drive across. It is a magnet to tourists, to 18

locals and of course to photographers from all over. It is one of the most photographed bridges in the world. Due to the situation of the Bay, almost every morning there is fog, which sometimes burns off during the day, other times there is haze or fog the entire day. Spectacular images of the fog rolling in in the evening are common in this great city. Meteorology plays an essential role in how much San Francisco Bay is photographed and the importance of the Bay to San Francisco.

Paris Now let’s look at how the Seine River in Paris enhances the city. Parisians love their river. They picnic, stroll, have meetings and parties on the banks or quays of the river. It is an enormous part of Parisian life. When taking a “Bâteau Mouche” up and down the Seine on a Saturday afternoon, it is easy to see a group of people dancing Salsa, a couple

eating bread and cheese and drinking wine, another pushing a baby carriage enjoying the afternoon. There are cyclists, joggers, sunbathersâ&#x20AC;Ślife happens on the quays of the Seine. The entire orientation of the city is talked about in terms of the river. You ask a Parisian where a specific building or place is, the answer is on the Right Bank or the Left Bank: the Rive Gauche and the Rive Droite and then they tell you the number of the Arrondissment. It may be confusing to some, but it is the way the river flows. When you face downriver the left bank is on your left and the right bank is on your right. As far as the arrondissements are concerned, they are administrative districts. The first arrondissement is on the Right Bank and considered the center of Paris- then the arrondissements are distributed in a spiral form like a snail.

Picture pag.18/19: Sunrise over the city of San Francisco. This picture was taken from the Twin Peaks. Map pag.19: Location of San Francisco in relation with the West Coast of the United States.

Picture pag.20/21: Panoramic photo of the city of Paris. The Pont des Arts (The Arts Bridge) can be seen accross the picture. Map pag.20: Lacation of Paris in relation with France and Europe.

Thirty-seven bridges connect Paris from one side of the Seine to the other. The Pont des Arts is a pedestrian bridge that crosses the Seine from the Institut Français to the Louvre. In recent years, it has become the romantic bridge of Paris. Tourist couples take padlocks with their names engraved on them and attach the paddocks to the railing or the grate on the sides of the bridge. Then they throw the keys into the Seine symbolizing eternal love. The Mirabeau Bridge is truly the melancholy bridge of Paris. It is so remembered because of Guillaume Apollinaire’s poem about the pain of love and the fatality of how time passes: “Under the Mirabeau Bridge flows the Seine And our loves Must I remember them Joy always followed pain The night falls and the hours ring The days go away I remain…” 20

Ever since Paris was founded the river has been vital to its inhabitants. Ever since photography was invented, the city of Paris with its incredible monuments, from the Tour Eiffel to Notre Dame Cathedral has been a true paradise for urban photographers. The Seine plays an enormous part in the majestic cityscapes of the City of Light.

London It is impossible to imagine London without the Thames River. It provides a splendid background for many of the architectural wonders of London such as the Parliament, Big Ben, the London Eye and the Tower Bridge. There are river cruises and tours up and down the river for tourists and Londoners alike. Walking, strolling and cycling are also favorite pastimes along the river. Riverside pubs and restaurants also attract crowds especially when the weather is good. The Thames has been an inspiration to artists, poets and photographers. There is sometimes fog and many times rain, which enriches the opportunities for extraordinary river and cityscapes for artists in general. Scores of famous artists have painted the Thames:

Canaletto, Monet and Pissarro just to mention a few. Whistlerâ&#x20AC;&#x2122;s paintings of scenes from the river shaped a large a large part of his work. The river and the cityscapes provided by the Thames have deepened the visual potential and imagination of artists and photographers over time. It is impossible to imagine any of the abovementioned cites without water. Rivers, oceans and bays of course are good reasons for having developed a small village into a city- commercial reasons being the foremost incentive. Water enhances commercial possibilities, but it also enhances life in the cities. Water is essential for our lives however not essential for all our activities, but water always gives a profounder meaning to our cities and to life. SueEllen Campbell, professor of nature and environmental literature puts it very clearly for many of us, â&#x20AC;&#x153;In my life, water has the character of punctuationâ&#x20AC;?. 21

A cityscape of London, England showing some of the most important tourist attractions and buildings.

Animals in Captivity:

Racehorses

Blanca Asperilla (exercise rider) training at El Hipodromo de la Zarzuela.

Racehorses at the Zarzuela Racetrack in Madrid

Picture pag.26: Exercise rider walking the horse before training, around the stables. Map pag26: Location of Madrid in relation with Spain and Europe. Picture pag.27: Horse being washed after training. 26

l Hip贸dromo de la Zarzuela or Zarzuela Racetrack was inaugurated in 1941 and ever since has been the center of horseracing in Madrid. The racing period in Madrid is from the end of February until the end of November (7). There is a very elegant restaurant there and it is a very popular venue to celebrate weddings and other social events. On the racing days (Sunday morning and Thursday evenings) tents are set up with refreshments so that the spectators can keep watching the races. On the racetrack grounds there are parks with swings and slides for children. The Zarzuela Racetrack makes a conscientious effort to keep all age groups active and occupied.

At present, the stables at the racetrack houses house 800 horses all year around. A racehorse is an animal that requires exquisite care and the dedication of many people to be successful. There are grooms to keep the stables clean, trainers, exercise riders and jockeys amongst others. Horses are not bred at this racetrack, they come when training begins. In October the new horses come. Training begins when the horse is around 18 months old. Racing begins at two years. Training at the beginning consists of trying to develop an emotional bond. First the trainer lets the horse get used to him by walking and petting the animal. At the beginning, the training of a racehorse is much like a normal horse. Slowly a halter is put on the horse, and then the horse is lounged and learns to stop and start again. Then the horse must be taught not to scare and then get used to bridle and a saddle.

Exercise Riders

The exercise rider rides and exercises the racehorse because trainers many times are too heavy. An exercise rider should not weigh more than 150 lbs. They begin with riding on a lounge line- and go from a walk to trot to canter and to gallop. We talked with one of the exercise riders at the Zarzuela Racetrack about her activities there. Blanca Asperilla has been an exercise rider for 4 years. She is 26 years old and is a Nurse. She has ridden since she was a child and has had a passion for horses since she can remember. She described her mornings at the racetrack as the most beautiful part of her day. She rides two racehorses called Ducati and Alacama. 27

Horse in stable at El Hipodromo de la Zarzuela.

Some workers taking care of one of the stables.

A normal day for Ms. Asperilla consists of arriving at the stables at 9:00 am. Dcucati is saddled and has had a little breakfast before going to the track. She takes him for an hour and a half workout. She says, “Ducati is a young horse full of energy and can’t wait to get out on the track. It takes about 20 minutes to get to the racetrack from the stables. I walk him, and then let him trot for 10 minutes, but all he can think about is galloping! We only let the horses gallop twice a week. He is a very nervous horse and scares easily. After exercising him, I ride back to the stable and the groom blankets him and I walk him to cool him down for about 15 minutes. I know the groom could do this but I enjoy this time with Ducati. We have a great relationship. He has a great personality and he even has a sense of humor. When I start daydreaming or walking slowly he nudges me in the back- once he almost knocked me over he is so frisky!”

Then this young exercise rider goes to the stable and does basically the same routine with Alacama. Ms. Asperilla has been riding Alacama since the beginning of his training and she talks about Alacama with love and admiration. He is an experienced racehorse and has won 3 national prizes. Ms Asperilla says, “Alacama and I have been through a lot together. He is very calm but a fast horse and he has an enormous heart. I do everything I should to keep him in line, well-exercised and welltrained and he has a trust in me that surpasses any other relationship with a horse I have ever had.” She goes on to explain, “Racehorses don´t exercise one day a week and sometimes I miss him so much on my day off that I swing by the stables just to say hi to him!.” 29

We took a walk around the stables and look at the beautiful horses. They are all groomed, well cared for and the stalls are clean. We ask Ms. Asperilla if she thinks that it is cruel to racehorses. She says, “This may sound crazy but these horses are super well taken cared of. I have heard stories of racetracks and stables that keep horses in awful conditions and don’t give them enough food, but I have ridden at practically every stable in Madrid and Zarzuela is the place with the absolute best conditions for horses I have ever seen in every sense. They are well fed; they have the very best medical attention with incredibly talented vets who adore the animals. Riders like me who love to ride and to care for them exercise the horses. When they have a physical problem they are taken to special premises where someone is with them 24 hours a day. And the question you asked if I think racing horses is cruel… In the conditions that I work, I certainly

don’t believe so! The atmosphere is wonderful with all my co-workers at the stables who have something to do with the horses and also amongst the people who come to the races. At the Zarzuela, going to the races is like a social gathering, some people bet, but it is not an ugly environment. Actually I think it is a pretty healthy atmosphere because most people who come are in some way connected to the world of horses and love them. Many people come here with the entire family every Sunday morning.” These horses are animals in captivity but are well taken cared of and loved by the people who surround them. We left the Zarzuela Racetrack after our guided tour by the young exercise rider with the feeling that a Sunday morning at the racetrack would be a delightful experience to have.

Blanca Asperilla (exercise rider) walking the horse after training.

Sperm about to be injected into egg during ICSI.

Small Worlds: IVF/ICSI

New Section

n 1978, Louise Brown, the first In vitro Fertilization baby was born. Just over 35 years have gone by and although Human Assisted Reproduction is far from perfect, great advances have been made (8). It is unimaginable for a fertility specialist in 2014 to think of performing egg retrieval transabdominally and without using some of the routine fertility drugs used in this day and age like the pregnancy achieved in 1978, but even so, human fertility continues to be very, very low.

Menstrual cycle and stimulation.

ovulation takes place and then the luteal phase begins. The uterus also has 3 phases: menstruation, proliferative phase where the uterine lining or endometrium grows and the secretory phase where the endometrium is transformed under the influence of progesterone (9). The first day of the cycle is the first day of menstruation. At the beginning of the cycle, estrogen levels starts rising and the endometrium thickens. Anywhere between 2 and 25 follicles (depending on the age and the ovarian reserve) begin growing and at one point one follicle becomes the dominant follicle and the other growing follicles become atresic. That follicle ruptures, a follicle is released and will live about 24 hours. The remains of the follicle in the ovary are called the corpus luteum, which produces important quantities of progesterone. If the egg is not fertilized the lining will shed, the corpus luteum starts to regress and a new cycle begins (9).

What kind of work goes on at a fertility clinic? The different ways of getting pregnant at a fertility clinic are: 1- Timed Intercourse: which consists of monitoring the cycle by ultrasound scanning and telling the couple when to have sex. 2- Intrauterine insemination or IUI: the cycle is monitored and 36 hours after one or 2 follicles have reached 17 mm and an hCG injection has been administrated the sperm is capacitated and injected into the uterus. The menstrual cycle is the series of changes that take place in a womanâ&#x20AC;&#x2122;s body every 21-35 days depending on the woman. These changes are indispensable for human procreation (9). The cycle of the ovary is made up of 3 phases: the follicular phase, where a number of follicles start to grow, a dominant follicle is selected and the second phase the follicle ruptures and 34

3- In vitro fertilization or IVF: the eggs are retrieved after stimulation and a drop of sperm is placed on the egg: 2, 3 or 5 days later 1, 2 or 3 embryos are placed in the uterus. 4- IVF with Intracytoplasmatic: conventional IVF with microinjection which consists in placing a spermatozoon inside an egg.

Small Worlds IVF/ICSI Although a fertility clinic does all these treatments, this article is going to focus on what happens in an IVF cycle.

How is an IVF performed? On Day+2 of the patient’s cycle, fertility medication is administered (FSH) to recruit all the follicles programmed to develop that cycle. The patient injects the shots every 24 hours and is monitored by ultrasound every day or two by the fertility expert. This process usually lasts an average of 10 days. When three follicles reach 17 mm, an hCG shot is administered and 36 hours later egg retrieval is performed (10).

How do we get more than one oocyte per cycle? A baby girl is born with an ovarian reserve of about 400 000 eggs. During a woman’s reproductive years between 2-25 follicles begin to grow every month and only one is ovulated per month. What happens in IVF is that from the second day of the menses FSH hormone injections are administered daily and all the follicles that were destined to grow will grow and due to the administration of FSH there will be no dominant follicle, instead the follicles should grow harmoniously (10).

The woman’s role: Egg Retrieval To begin the egg retrieval the needle used to extract the eggs is purged with culture medium, which is oocyte-friendly. Then the extraction is begun. The doctor pierces each follicle and extracts the follicular fluid and the egg. This fluid and egg end up in a test-tube through a suction system. When the tube is filled it is taken to the

embryologist in the IVF Lab who empties the contents into a plate and looks for oocytes with a microscope under a sterile hood. When the embryologist finds an oocyte, it is placed into another plate with culture medium. At the end of the egg retrieval the embryologist reviews all the plates where the follicular fluid and eggs have been emptied to assure that no eggs have been left behind. The eggs are placed into an incubator and are kept there for 3 hours so they can stabilize after the traumatic process it must be to be aspirated out of a follicle. (Picture 1 and 2, pag. 38) While the eggs are stabilizing in the incubator in the Embryology Lab, the sperm sample is being processed in a procedure called capacitation (11).

The man’s role: Sperm Collection This takes place in the Andrology Lab. The objective of capacitation is to separate cell detritus, seminal fluid and spermatozoa that are not optimal from the motile and normal sperm. This enriched concentration of spermatozoa is left to complete capacitation in culture medium (IVF. Vitrolife®), a culture medium friendly to spermatozoa. This technique is called swim-up as the best spermatozoa are the ones that are going to swim upwards after the sample has been centrifuged. Though in this process the spermatozoa are going to suffer, the same capacitation process would have taken place as the sperm swam through the female reproductive organs. (Picture 4 pag.39)

Graphic pag.34: It shows the different stages of the menstrual cycle. It shows how hormones, ovarian histology and the endometrian histology change depending on the day of the menstrual cycle, showed under all the drawings. 35

New Section Mixing time! Conventional IVF or Fertilization by ICSI (Day 0) About 2-3 hours after egg retrieval the eggs or oocytes are denuded. In this process the oocyte is stripped from the oocyte-cumulous complex (formed by the oocyte and the surrounding cells, called granulose cells, which are the ones that “feed” the oocyte during its maturation in the ovary. Once the oocytes are denuded the number of the Metaphase II oocytes (mature oocytes) are known. These oocytes are ready to be microinjected (12). (Picture 3 pag.39). Another way to fertilize the oocyte is by conventional IVF, which means that a drop of capacitated sperm is placed on top of the egg allowing nature to determine which spermatozoon will enter and fertilize the egg. If the sperm and egg are both good, fertilization will be observed between 18-22 hours later (12).

and 2 nuclei (one corresponding to the mother and the other to the father). If the egg is not correctly fertilized sometimes it can show more than 2 nuclei. This event can be due to 2 different causes: polispermia (fertilization of the egg by 2 different spermatozoa, related with the inability of the hardening of the zona pellucida when the first sperm penetrates into the oocyte), or genetic abnormalities in the oocyte. (Picture 5 pag. 38).

Evaluating embryo development. D+2, D+3, D+4, D+5 Every day afterwards, the embryos are taken out for observation and their quality is recorded. On Day+2 four cells are seen. They should be equal in size and no fragmentation should be observed. Embryo quality is the parameter used when selecting the embryo/embryos to transfer. There are many classifications but in general the following criteria is used when qualifying embryos.

At present, most labs use microinjection or ICSI (Intra-Cytoplasmatic Sperm Injection) to fertilize oocytes because the fertilization rate is higher. Grasping the oocyte with the holding pipette, ICSI is performed. A spermatozoon, with normal morphology and that is swimming straight and fast is located in the ICSI Petri dish. The tail of the sperm is cut off and then is aspirated by the microinjection pipette. The oocyte’s polar body is placed at 6 o’clock and then the sperm is inserted into the egg. The gametes are then placed in culture medium and the put into a conventional incubator or the EmbryScope, a state of the art incubator.

- Day 2 and 3 embryos are classified according to their morphology. At this moment the number, position, appearance of cells and presence/lack of fragmentation is evaluated.

Evaluating fertilization. D+1

Embryos achieve the blastocyst stage when on Day+5 or 6. Blastocysts are more complex in their structure, first of all because because it has many more cells but the vital difference is that these cells are differentiated into 2 cell types. The first cell type is the trophoectoderm, which will later become

When using a conventional incubator, the gametes are taken out 22 hours later for observation. At this point fertilization should be observed. A correctly fertilized oocyte should have 2 polar bodies 36

- On day 2, a well developing embryo should have 4 even cells with no fragmentation and it should duplicate its cells day by day, showing 8 cells on day 3 and starting compaction on day 4. (Picture 5 and 6 pag. 38). - On day 4, the developing embryo is called morula (12). (Picture 7 pag. 39).

Small Worlds the placenta and the inner cell mass, which will give rise to the fetus. A blastocyst also has a central cavity full of liquid, which is called the blastocoel. (Picture 8 pag. 39). The most popular classification system for blastocysts is with Gardner’s Classification (5) which takes into account the expansion of the blastocell (graded 1,2,3,4,5); inner cell mass (graded A,B,C) and the trophectoderm (graded A,B,C) The most advanced and highest valued blastocyst is a hatching blastocyst (graded 5). The blastocyst makes a hole in the zona pellucida and starts to look to implant.

The Embryo Transfer: D+2, D+3 or D5 Embryos can be replaced o Day+2, Day+3 or Day+5 depending on many factors such as age, sperm and egg quality and the personal history and circumstances of the patient. - On Day+2: 4 cells equal in size should be visualized with no fragmentation. - On Day+3: 8 cells should be counted. They again should be equal and in optimal conditions no fragmentation should be observed. - On Day+4: the embryo reaches the morula stage where the morphology of the embryo is like a compact ball that then over the following 24-48 hours expands into a blastocyst. - The former description is what the embryologist observes when embryos are cultured in a conventional incubator. If after microinjection the gametes are placed in the EmbryoScope® embryo observation is spectacular because of the incorporated time-lapse photography mechanism the EmbryoScope® has. The growing embryos

have their picture taken every 7 minutes 24 hours a day. This allows the embryologist to see the exact development of the embryos and their growth patterns. When the images are looked at in sequence, a video of the growth of the embryo is obtained. This gives the embryologist much more information to select the best embryo to transfer.

Goal of Assisted Reproduction Techniques The goal of Assisted Reproduction treatments at present is one healthy take-home baby. For that reason Single Embryo Transfer is gaining more momentum because although “Time to Pregnancy” can be longer, pregnancy rate is the same and the multiple birth possibility is almost at 0%. Even when one embryo is transferred it can split causing the patient to have identical twins. After selecting the embryo/s to transfer the biologist must load the catheter where the embryo/s are to be placed for embryo transfer. This is done at the moment when the doctor has placed a catheter into the uterus of the patient. The embryo/s are loaded in 20 microliters of culture medium and are left between 10-20 mm from the fundus of the uterus. To do this the embryo transfer is performed using abdominal ultrasound and thus the embryo/s are placed at an optimal location for implantation. The pregnancy test is done 12 days after blastocyst transfer checking the -hCG blood levels. There are many periods in the IVF process, many steps where near perfection is needed to achieve pregnancy. It is truly a small world where expertise, utmost care and the highest respect for life is needed to succeed.

Picture 2: Retrived eggs.

Picture 1: Retrived egg.

ICSI

Timeline

Picture 5: Day +2 of fertilization (4 cells).

Picture 6: Day +3 of fertilization (8 cells).

Picture 3: Eggs after being denuded.

Picture 4: Sperm after capacitation.

Picture 7: Day+4 of fertilization (Morula stage).

Picture 8: Day +5 or 6 (Blastocyst).

Urban Wildlife: Turtles in Atocha

A view of the Atocha Railroad Station in Madrid, Spain.

URBAN WILDLIFE Exotic turtles in the Atocha Railroad Station in Madrid, Spain Spain is a country of great contrasts. Climates vary from Mediterranean to tropical, there are different languages in the north and the west of Spain, different accents in just about every province, different cuisines in each region, just to mention a few of the many disparities. One of the oddest contrasts in Spain exists at the Madrid Atocha Railway Station in Madrid. Atocha is the largest station in Madrid and the principal commuter train station, the intercity and regional station for the cities of southern Spain and it is home of the AVE (abbreviation for Alta Velocidad Española or Spanish High Speed Train). The high-speed trains go to Barcelona, Zaragoza, 42

Seville and Valencia. Atocha Train Station was first constructed in 1829 but over the years, as the needs evolved the station’s architecture changed. The last big makeover was in 1992 when the tracks for the high-speed train were built in an additional building. The old station’s tracks were pulled up in the original and main building leaving an enormous space. The incredible idea of designing an enormous tropical garden for the empty space was consolidated. The 4000 square meter space is filled with 7200 plants made up of 260 species (13) The astonishing contrast is that the highspeed train is housed next to the exotic tropical garden, which has a pond. In that pond, there are more than 300 turtles, one of the slowest moving reptiles. The high-speed train’s maximum speed is 300 km/hour and a turtle takes between 20-50 minutes to cover 100 meters (14)!

How could 300 turtles end up in a pond in a train station? Was this a planned strategy? According to a spokesperson from ADIF (Administrador de Infraestructuras Ferroviarias, the Administrator of Railway Infrastructure) this pond was not designed for turtles but for aquatic plants, but citizens who did not know what to do with their turtles started bringing them to the pond and released them there. As people traveled by AVE and saw the pond, they begin to think that anybody could let their unwanted turtles go and the pond started to fill. Another factor that has contributed to the growing turtle population at the station is that in Spain, if you want to travel by train with any animal, you have to pay 40% of an adult ticket (15). Some people get to the station and then donâ&#x20AC;&#x2122;t have the means to pay for the ticket and have to opt for releasing their pet turtles as a last minute solution.

Picture pag.42: Travellers taking a look at the pond where the turtles are enjoying some sun. Map pag.43: Location of Madrid in relation with Spain and the rest of Europe. Atocha train station is located in Madrid. 43

Even though it was not conceived as a turtle pond, it is a beautiful venue for one. The magnificently high glass ceilings, which let the sunâ&#x20AC;&#x2122;s warmth and light shine through the open space, the warm atmosphere surrounded by a tropical garden with palm trees and ferns all come together in harmony. The tropical garden has an hourly humidifying spray to keep the environment favorable for the tropical plants. This is a perfect habitat for the turtles who thrive even though they pond is small. In the pond there are many species: from native Spanish turtles to exotic turtles from North America, (the northern map turtle (Graptemys geographica) and the painted turtle (Chrysemys picta)), from Asia (the Chinese softshell turtle, Pelodiscus sinensis) and even turtles from Africa like the Pelomedusa Subrufa. 44

The spokesperson underlined the fact that turtle releasing is not encouraged and people should realize that when they buy a turtle it is going to grow. At the same time the turtles are fed daily. Actually, at present, the turtle pond is one of the biggest attractions at the Atocha Station. Aurora PeĂąa, who takes care of the turtle pond (15), has what appears to be an enormous butterfly net. After feeding the turtles daily, she spends some time fishing out objects like sunglasses that the travellers have accidently dropped into the pond. She also comments on how there is a space with sand so the turtles can lay their eggs and take care of them until they hatch. Even though the practice of releasing these domestic and exotic turtles in the Atocha pond is not encouraged, it is tolerated. These ponds have popped up in another 4 locations in Madrid. The

problem is that one of the other options for turtle owners who tire of their pets have, is to let their exotic turtles go in the wetlands in different locations throughout Spain. This is devastating because these exotic species are predators of the autochthonous wildlife of the region. An example is the Wetlands of Bolue in the northern part of Spain, where each year more than 50 exotic turtles are collected. These turtles have been extremely harmful to the environment and the authorities are actively trying to eradicate them (15). Each big city has unexpected types of urban wildlife. These exotic and native turtles living together in harmony at a train station in Madrid are certainly a remarkable example of urban wildlife. Picture pag.44: Turtles enjoyig the sun one on top of the other. Picture pag.45 (large): Pigeon visiting the turtles in the station. Picture pag.45 (small): A lot of turtles enjoying the sun in one of the spaces prepared for them.

&Maps

Bibliography

1. “Carmel Mythbusters.” Aol News. Aol Travel, n.d. Web. 4 May 2014. <http://news.travel.aol. com/2010/09/07/carmel-mythbusters/>. 2. Point Lobos Foundation. N.p., n.d. Web. 4 May 2014. <http://www.pointlobos.org/nature/birds>. 3. Roberson, Don. Monterey Birds. N.p.: MPAS, 1985. Print. 4. The IUCN Red List of Threatened Species. N.p., n.d. Web. 1 May 2014. <http://www.iucnredlist.org/ details/22696753/0>. 5. Fix, David, and Andy Bezener. Birds of Northern California. N.p.: Lone Pine, 1957. Print. 6. LaDeau, Shannon L.; Kilpatrick, A. Marm and Marra, Peter P. (2007). “West Nile virus emergence and large-scale declines of North American bird populations”. Nature 447 (7145): 710–713. doi:10.1038/nature05829. PMID 17507930. 7. Hipodromo de la Zarzuela. N.p., n.d. Web. 4 May 2014. <1.http://www.hipodromodelazarzuela.es/ historia/historia-del-hipodromo-de-la-zarzuela> 8. BBC News. N.p., n.d. Web. 1 May 2014. <http:// news.bbc.co.uk/2/hi/health/3077913.stm>. 9. Speroff, Leon, and Marc A. Fritz. Endocrinología Ginecológica Clínica y Esterilidad. 7th ed. N.p.:Lippincott Williams and Wilkins, 2006. Print. 10. Remohí, J., et al. Reproducción Humana. 2nd ed. Madrid: McGRAW HILL, 2002. Print. 11. Matorras, Roberto. Estudio y Treatamiento de la Pareja Esteril. Madrid: n.p., 2007. Print. 12. Gardner, David K., Ariel Weissman, and Colin M. Howless, eds. Textbook of Assisted Reproductive Teachnologies. 3rd ed. N.p.: n.p., 2004. Print. 13. Madrid Diario. N.p., n.d. Web. 2 May 2014. <http://www.madridiario.es/>. 14. lainformacion.com. N.p., n.d. Web. 4 May 2014. <http://noticias.lainformacion.com/asuntos-sociales/ la-estacion-de-tren-de-atocha-el-asilo-de-las-tortugasabandonadas_XZJZ4cNMUUWnKR0GDNoFw3/>. 15. El País. N.p., n.d. Web. 4 May 2014. <http://ccaa. elpais.com/ccaa/2013/07/08/madrid/1373311089_345874. html>.

The Peak District. Latitude 53.2 Longitude -1.67 Picture from Contents was taken here. Mindo, Ecuador. Latitude -0.05 Longitude -78.7 Picture from Editorial was taken here. Carmel by the Sea, CA, US Latitude 36.5 Longitude -121.9 All the pictures from the bird article were taken here. Seville, Spain. Latitude 37.3 Longitude -5.98 Pic 1 from the Water and City Life article was taken here. Miami, FL, US. Latitude 25.78 Longitude -80.2 Pic 2 from the Water and City Life article was taken here. San Francisco, CA, US. Latitude 37.7 Longitude -122.4 Pic 3 from the Water and City Life article was taken here. Paris, France. Latitude 48.8 Longitude 2.35 Pic 4 from the Water and City Life article was taken here. London, UK. Latitude 51,5 Longitude -0.11 Pic 5 from the Water and City Life article was taken here. Madrid, Spain. Latitude 40.45 Longitude 3.7 All pictures from Racehorses, Small Worlds and Urban Wildlife were taken here.

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Birds Carmel by the Sea ISO 100 300 mm f/5.6 1/13 sec

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Urban Wildlife Turtles in Madrid

Landscapes Water and City Life ISO 100 18 mm f/4.0 1/1600 sec ISO 100 85 mm f/4.0 1/2000 sec

Animals in Captivity Racehorses

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