“This time results of an unlimited license granted to technology” and.... “constitute the perfect crime to kill reality” Jean Baudrillard

Andres E. Carrasco Laboratorio Embriologia Molecular Facultad de Medicina UBA-CONICET

Argentina and agrotoxics

and Territorial conflicts

El futuro:

P.A.N

Extension: 20.000.000 Ha. GBH: 200.000.000 liters year , (Paraguay GBH 8 liters year) Population living in the territory 10.000.000 people.

70 % of the effects are by inhalation.

Soy in Argentina

Increasing of use of plaguicides 1999-2009

Preliminary epidemiological humans studies in Argentina

Birth prevalence of 27 selected congenital anomalies in 7 geographic regions of Argentina ,

Campaña et al (ECLAMC group). Arch Argent. Pediatr. 108 (5) 409-417, 2010

Out of the 27 congenital anomalies analyzed, 14 showed a frequency significatively higher in one or more regions

Centro spina bifida, microtia, cleft lip with cleft palate, polycystic kidney, postaxial polydactyly and Down Syndrome, Northest omphalocele, gastroschisis, cleft lip with and without cleft palate, anorectal atresia/stenosis, indeterminate sex, preaxial polydatily and pectoral agenesis,

Patagonia cleft lip with cleft palate. Cuyo postaxial polydactyly, Pampa, severe hypospadias

CORDOBA

Obstetric and Neonatology Hospital, Cordoba Univ, (Dra. Trombotto)

On 111.000 newborns,

CRANEOFACIAL , GASTROSCHISIS, LIMB DEFORMITIES

Incidence of MCM (not due to genetics or familiar bias)

1991: 16.2 %o newborns. 2003 : 37.1 %o newborns.

Ituzango, Cordoba.

RED DOTS: CANCER CASES

Departamento Bermejo

Puerto Eva Perón

Puerto Bermejo

General Vedia

LA LEONESA

Las Palmas

Isla del Cerrito

Chaco, Argentina

Municipio Población

Total 2001

Menores de 15 años

- 2001

Casos Esperados Ca Infantil - por año

Casos Registrados

Incidencia años

La Leonesa 10.067 2960

0,41 casos por año ( 1 caso c/ 24- 36

meses)

1995: 1 caso 1997: 1 “ 2000: 1 ” 2003: 2 “ 2004: 1 “ 2008: 1 “ 2009: 1 “

1990-1999 0,2 casos/año – 1 caso/60meses

2000-2009 0,6 casos/año – 1 caso/20 meses

Las Palmas 6593 2146

0,3 casos por año ( 1 caso c/ 36-42

meses)

1993: 1 “ 1995: 1 “ 2006: 1 “

1990-1999 0,2 casos/año 1 caso/60 meses

2000-2009 0,1 caso/año 1 caso/ 120 meses

Puerto Bermejo 1832 652

0,09 casos x año ( 1 caso c/ 96

años)

1995: 1 caso 2008: 1 “

1990-1999 0,1 caso/año 1 caso/20 meses

2000-2009 0,1 caso/año 1 caso/120 meses

La Leonesa, Dept. Rio Bermejo, Chaco

300%  

Malformations % in total live newborns

Año N /year Live born Incidency (malformations *10000/

live born ) 1997 46 malformacions 24030 (live born 1997) 19,1 /10.000

2001 60 malformacions 21339 (live born 2001) 28,1 /10.000

2008 186 malformacions 21.808 (live born 2008) 85,3 /10.000

400%  en  los  ul,mos  10  años      

Benitez Leite et al, Rev Ped. Paraguay (2009)

139 newborns were analyzed. Where founded 52 malformations associated to exposure to agrotoxics (plus 34 death fetus).

In Roque Saenz Peña, Chaco, Argentine. In 4 years, over 4255 of newborns, 157 malformations were detected (average 3.6%)

Documents:

CONICET (2009)

UNIV. DEL LITORAL (2010)

GM SOY: SUSTAINABLE? RESPONSIBLE? (2010)

ROUNDUP and BIRTH DEFECTS (2011)

PESTICIDES USED IN SOUTH AMERICAN GMO-BASED AGRICULTURE: A REVIEW OF THEIR EFFECTS ON HUMANS AND ANIMAL MODELS

Silvia L. López; Delia Aiassa; Stella Benítez-Leite; Rafael Lajmanovich; Fernando Mañas; Gisela Poletta; Norma Sánchez; María Fernanda Simoniello; and Andrés E. Carrasco.

Advances in Molecular Toxicology Vol 6, 2012, (in press)

Legal actions to block effects of agrochemicals

San Jorge, Santa Fe.

Sentence: 800 m. for ground machine and 1500 m. aerial aspersion. Legal action. Santa Fe´s Court of Apppels.

La Leonesa, Chaco.

Sentence: 1000 m. for ground machine and 2000 m. aerial aspersion. Legal action. Chaco’s Superior Court .

National Ombusman.

Resolution recommending to the Minister of Agriculture to review classification of all agrochemicals used in Argentina.

Glyphosate and teratogenesis

Experimental evidences of TOXICITY of AGROCHEMICAL

GLYPHOSATE

 BLOCK THE DNA REPAIR MACHINERY. (checkpoint enzymes)

 BLOCK CELL CYCLE DURING DEVELOMENT.

 ACTIVATE APOPTOSIS IN CELL CULTURE.

 PRODUCE TERATOGENESIS DURING VERTEBRATE DEVELOMENT (AVIAN, ANPHIBIAN, MAMMALIANS)

Laboratorio de Embriología Molecular Facultad de Medicina

Universidad de Buenos Aires – CONICET  

acarrasco@fmed.uba.ar  andresecarrasco.blogspot.com  

6th  European  Conference  of  GMO-­‐Free  Regions  Brussels  and  Ghent    16-­‐17  September,  2010    

ARGE  Gentechnik-­‐frei,  Soy?  –GMO-­‐free  &  Sustainable    Vienna,  April  28  2011  

Two types of experiments were designed :

1.  Embryos were cultured saline solution (BM) with 1/5000 dilution of GBH (430 uM)

2.  200 to 300 pg of glyphosate (in 4.6nl) was injected in one blastomere of the two cell embryo (final concentration in the injected cell was 10 uM)

Izq Der

Injection volume represent 1/220 fold the volume of the injected egg (1ul)

4.6nl

1.  Microcephaly 2.  Microphthalmia 3.  Craniofacial disruption

(branchial arches) .

5.  Shortening of anterior posterior (AP) axis.

Both GBM and glyphosate injection alter midline formation by down regulation of Shh and Pax6 In tadpoles down regulation of Otx2 and Sox9 produces:

Paganelli, et al 2010

1.  Microcephaly 2.  Microftalmia 3.  Craniofacial cartilages 4.  Shortening of Anterior posterior

axis. 5.  Ciclopia and Holoprosencephaly

Paganelli,  et  al  2010  

In larvae stage both GBH and glyphosate produce disruption of development craniofacial due to inhibition of regulation of genes expressed in the embryonic midline (Shh, Otx2)

• GBH increase retinoic acid activity. • RA antagonist (Ro) rescue the GBH induced phenotype.

Paganelli,  et  al,  2010  

Retinoic acid (a derivate of vitamin A) plays an important role in the regulation of expression of genes controlling early embryogenesis in vertebrates.

Retinoic acid is an important morphogen for the pattern formation of three embryonic axis

Glyphosate    Synthesis  or  degradaNon    (RALDH  or  cytochrome  P450    CYP  26  A,  B  y  C)  

Retinoic acid

HOX    

Otx2     Shh  (HNF3B)  

FGF  8    

Cranial  neural  crest    Otocephaly   HPE  and  cyclopia    

anterior-­‐posterior  axis  

Anencephaly  

In  conclusion  

Retinoic acid

Retinoic Acid

 RA is a diffusible morphogen present in vertebrates that biding a family of nuclear receptors (RAR), regulating early genes during gastrulation (3 week of pregnancy) and late morphogenesis.

 RA regulates formation of AP embryonic axis

 In adults play role in fertility, vision, prevents neoplastic growth, and neurodegenerative disease.

RA excess cause 20 % of the abortions and 35 % of the malformations during embryonic pattern

formation

RA is ligand for a nuclear receptor and gene regulator

Pera et al 2009

Retinoic Acid gradients in the embryonic A/P axis

Human syndromes caused by excess of RA

(excess of vitamin A)

CRS: Caudal Regression Syndrome in mice. (excess of RA) (Padmanabhan R., Retinoic Acid-induced caudal regression syndrome in the mouse fetus, Reproductive Toxicology, 1998)

Exencephaly. Microcephaly, Microphthalmia. Exophthalmia. Holoprosencephaly,(HPE) Spina bífida y mielomenigocele

Agnatia or micrognatia, aglosia, palate agenesis, cleft palate craniofacial abnormalities. Astomia (mouth agenesis) and Anotia (auricular agenesis),

Imperforate anus, Rectouretral fistula , Kidney malformations or kidney agenesis, Criptoquidia. Gastroschisis,

Reduction ribs number No tail and caudal vertebral defects, Limb malformations (meromelia, sirenomelia* (see picture) and syndactily)

Pathological morphogenesis of heart, lung, kidney, pancreas)

Caudal Regression Syndrome could be explained as:

  A disorder of embryonic midline formation results in the inhibition of several regulatory pathways (Hox complex, A/P axis, Otx2, Shh, FGF8, etc.).

  Functional disruption of developmental mechanisms: proliferation, apoptosis and cell migration during early vertebrate morphogenesis.

Retinoic acid syndrome

Inhibition of Otx2 (forebrain and midbrain) HPE (ciclopia and prosbosis) + agnatia and sinotia

Inhibition of Shh (embryonic midline) HPE, situs inversus, limbs (amelia, sirenomelia, sindactily, etc.)

Missregulation of Hox genes (affects A/P axis, vertebral identity and hindbrain)

Malformations: lung, diafragm, eyes , hearth, digestive tract, kidney, gonades, etc.

NEURAL CREST

NEURAL CREST

Scientific reports associate chemicals and abnormal morphogenesis for retinoic acid pathway disruption.

Atrazine Lenkowski J, et al. Env Health Persp (2008, 2010)

Tridimefon, Papis E, et al. Gene expression patterns (2007)

Arsenic, Davey C, et al. Env Health Persp (2008)

Alcohol, Kot-Leibovich H, et al. Dis Model Mech (2009)

Azoles, Giavini E. and Menegola E., Tox. Letters (2010)

Glyphosate in mammalian embryos

Doses: IV 100 mg/Kg Oral 400 mg/kg

Slowly disappearance from blood

Intravenous half life: 10 hr. Oral half life: 14 hr (degradation to AMPA 6%)

Distribution (two compartment model) both administrations indicates that glyphosate penetrate easily all tissues (according with Brewster et al 1991)

Anadon, et al, Toxicology letter (2009)

Toxicokinetics of glyphosate

7 days

10 days

14 days

Human Embryo 14 days

Implantation in humans

Wrong!!!!!! Placenta is permeable to glyphosate. 15 a 20 % of glyphosate in blood, cross the placenta barrier.

“These experimental conditions do not reflect human exposure, kinetics (absorption, distribution, excretion and their time course) in the mammalian organism or factors such as the placenta barrier in the pregnant female. In no case, exposure of the developing human fetus in utero would be similar to that one of the frog or chicken embryos in the study by Paganelli et al.” (page 3-4)

Bundesant fur Verbraucherschutz und Lebenmittelsicherheit 19 October 2010

Dallegrave et al, 2003 LD50 5.6 g/kg glyphosate LD50 1g/kg POEA

Roundup (Brasil) 38% glyphosate 18% POEA

In mammalians (rats). Teratogenesis at 3.5 g/kg (acute oral LD50: 5.6 g/kg)

They used: 1 g/kg 075 g/kg 0.5 g/kg

Administration 9 days/pc Gastrulation 7 days/pc

In rats:

More information see: “Reply to the letter to the Editor Regarding our article” Chem. Res. in Toxicology, 24, 2011, 610-615

Comments

treatment was done after late gastrulation when the primitive streak and the axis are already setup.

craniofacial malformations

malformation in the skeleton (vertebrae, ribs, limbs, femur, etc)

All of these malformations are consistent with the increase of RA during development.

(see also “Reply to the letter to the Editor Regarding our article” Chem. Res. in Toxicology, 24, 2011, 610-615)

Persistency of glyphosate in the soil

New evidences of Roundup (glyphosate formulation) impact on the periphyton community and the water quality of freshwater ecosystems. Marıa S. Vera, Leonardo Lagomarsino, Matıas Sylvester, Gonzalo L. Perez. Patricia Rodrıguez, Herna´n Mugni, Rodrigo Sinistro, Marcela Ferraro, Carlos Bonetto, Horacio Zagarese, Haydee Pizarro

Ecotoxicology (2010) 19:710–721

“Unexpectedly, we detected that a single application of glyphosate in 2005 shifted the mesocosms from a ‘‘clear’’ to a ‘‘turbid’’ state which remained until the next year. As was discussed above, the glyphosate may be adsorbed to sediments and a slow later desorption might produce a long turn effect suppressing growth of the most sensitive groups and favoring the abilities to compete of the more resistant algae. This trend in long term effect was suggested by Holtby and Baillie (1989) who reported an enhancement of periphytic production as a response to increased levels of phosphorus produced by a unique application of Roundup done 1 year before their experiment, carried out in natural streams”

GENOTOXICITY

Type B Type D Type F

Type A (normal) Type C Type E

DNA Fragmentation

(Comet assay ) GENOTOXICITY

Genotoxicity (comet assay) (Rio Cuarto, CORDOBA)

Mañas y Aiassa 2010

PARAGUAY

PARAGUAY

Benitez Leite 2010