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Food Additives & Contaminants: Part A Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/tfac20 Residues of medroxyprogesterone acetate detected in sows at a slaughterhouse, Madagascar Vincent Porphyrea, Michel Rakotoharinomeb, Tantely Randriamparanyc, Damien Pognona, Stéphanie Prévostd & Bruno Le Bizecd a Centre de Coopération Internationale en Recherche Agronomique pour le Développement – CIRAD, Réunion, France b Direction of Veterinary Services, Ministry of Livestock Production, Antananarivo, Madagascar c National Laboratory of Veterinary Diagnostic, Antananarivo, Madagascar d Laboratoire d’Etude des Résidus et Contaminants dans les Aliments (LABERCA), Ecole Nationale Vétérinaire, Agroalimentaire et de l’Alimentation Nantes Atlantique (ONIRIS), Nantes, France Accepted author version posted online: 24 Sep 2013.Published online: 20 Nov 2013.

To cite this article: Vincent Porphyre, Michel Rakotoharinome, Tantely Randriamparany, Damien Pognon, Stéphanie Prévost & Bruno Le Bizec , Food Additives & Contaminants: Part A (2013): Residues of medroxyprogesterone acetate detected in sows at a slaughterhouse, Madagascar, Food Additives & Contaminants: Part A, DOI: 10.1080/19440049.2013.848293 To link to this article: http://dx.doi.org/10.1080/19440049.2013.848293

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Residues of medroxyprogesterone acetate detected in sows at a slaughterhouse, Madagascar Vincent Porphyrea*, Michel Rakotoharinomeb, Tantely Randriamparanyc, Damien Pognona, Stéphanie Prévostd and Bruno Le Bizecd aCentre de Coopération Internationale en Recherche Agronomique pour le Développement – CIRAD, Réunion, France; bDirection of Veterinary Services, Ministry of Livestock Production, Antananarivo, Madagascar; cNational Laboratory of Veterinary Diagnostic, Antananarivo, Madagascar; dLaboratoire d’Etude des Résidus et Contaminants dans les Aliments (LABERCA), Ecole Nationale Vétérinaire, Agroalimentaire et de l’Alimentation Nantes Atlantique (ONIRIS), Nantes, France (Received 4 July 2013; accepted 20 September 2013)

In Madagascar, little information about drug residues in animal products is available. However, recently, official veterinary services were informed about the misuse of human injectable contraceptives in pig farms as an alternative for chirurgical castration of adult sows before culling. We investigated pigs (n = 80) slaughtered in 7 Malagasy abattoirs and raised in 8 of the 22 Malagasy regions (1) to confirm the contamination of carcasses by anabolic hormones by using LC-MS/MS, (2) to identify the substances of concern and (3) to explore the consumers’ exposure to hormone residues. Medroxyprogesterone acetate was the only synthetic hormone detected in kidney fat. Samples positive with medroxyprogesterone acetate were observed in 66.7% of the districts investigated and in 87.5% of the surveyed regions, confirming its large misuse in livestock. Public awareness campaigns and control improvement among the animal production sector and among the Malagasy public health sector are therefore urgent. Keywords: pork; kidney fat; food safety; growth promoters; anabolic; medroxyprogesterone acetate; Madagascar

Introduction systems in developed countries allow an efﬁcient and Synthetic progestagens have been widely used in gynae- rapid alert to protect consumers, the situation is unclear cologic practice. Progestin medroxyprogesterone acetate in most of the developing countries regarding the use of (MPA) is used in contraceptive medication and also in hormones in animal farms and the hazard exposure for human hormone replacement therapy, but its side effects local consumers. are commonly reported: MPA has a signiﬁcant glucocorti- In Madagascar, the consumption of pork meat is tradi- coid activity that decreases bone density (Ishida & tional and the demand for pork meat is increasing in urban Heersche 2002), and it may also impair memory (Braden markets. The pig production sector is extensively devel- et al. 2010). MPA is associated with the development and oped with complex trade circuits from remote rural areas maintenance of obesity, and it may also be considered as a to urban abattoirs and retailing markets. In pig farms, few risk factor for cardiovascular diseases (Clark et al. 2005). biosecurity measures are implemented (Costard et al. In animal husbandry, MPA is allowed for reproduction 2009), leading to numerous food safety issues in pork purpose in sheep. MPA also possesses anabolic activity meat (Rakotoharinome et al. 2012; Temmam et al. 2013). In particular, the misuse of hormones in pig farms

Downloaded by [Cirad-Dist Bib Lavalette] at 10:04 22 November 2013 and therefore has been used in livestock production to improve muscle gain (Meyer 2001). Within the European has been increasingly reported during 2012 by local veter- ﬁ Union, the use of progestagens and other steroid hormones inary of cers (personal communication, M. for the purpose of fattening farmed animals is prohibited Rakotoharinome, Feb 15, 2012). Farmers and local animal (Courtheyn et al. 2002). According to the European health workers were suspected of treating pigs for promot- Commission Decision 2003/181/EC, the minimum ing their growth and of using human progestins, especially required performance limit in animal tissues is equal to MPA, as a chemical alternative method for the castration 1µgkg−1 (Commission of the European Communities of sows that are then fattened before culling. Indeed, MPA 2003). may arrest the onset of farrowing, and induce post-lacta- In European Union countries, the misuse of synthetic tional anoestrus in sows (van Leengoed et al. 2002). hormones in animal husbandry was reported in milk Because the use of synthetic hormones in pig husbandry (Unusan 2008) and pork meat (Oksbjerg et al. 1995). is considered as a fraud under Malagasy regulations MPA-contaminated feed was also incriminated (van (Malagasy Ministry of Agriculture 2006), an exploratory ﬁ Leengoed et al. 2002). While veterinary monitoring study was carried out to con rm these suspicions and investigate the main substances concerned.

*Corresponding author. Email: [email protected]

Material and methods Table 1. List of components, and their associated individual α −1 Field data limit for decision (CC ) (µg kg ), investigated in kidney fat by a multi-residues method by LC coupled with a quadripole The study considered 8 of the 22 regions of Madagascar mass spectrometer. where suspicions were reported by local field veterinarians: CCα* screening CCα confirmation Bongolava, Vakinankaratra, Itasy and Haute Matsiatra in Analyte (µg kg−1) (µg kg−1) Central Madagascar; Betsiboka, Diana and Sofiainthe Northern region; and Anosy in the Southern region. Flugestone acetate 0.27 0.39 Samples were collected during October 2012 from two pro- Norgestomet 0.29 0.56 vincial abattoirs, that is, Tsiroanomandidy in Western Delmadinone acetate 0.41 1.00 Altrenogest 0.19 0.38 Madagascar and Antsirabe city in Vakinankaratra province Chlormadinone 0.77 0.88 (Central Madagascar), and from four abattoirs in acetate Antananarivo, the Malagasy capital city, where animals Medroxyprogesterone 0.23 0.53 from all over the country were transported for slaughtering. acetate Nineteen communes from 15 districts within these eight Melengestrol acetate 0.74 2.41 regions were consequently sampled. Only adult sows were *CCα is defined as the limit of concentration from which a sample is sampled considering that chemical castration with progesta- declared not standard, at screening and confirmation steps, with an error risk equal to α. Here, CCα for medroxyprogesterone acetate, at the gens was the most common hypothesis of the misuse of – confirmation step, is lower than 1 µg kg 1 considered as the minimum synthetic hormones in the field. Eighty kidney fat samples required performance limit. were collected after slaughtering by veterinary officers and stored at −80°C before testing. Samples were shipped in dry 0.5 µg kg−1. Table 1 details the chemical constituents ice at −80°C from Madagascar to Nantes (France). Samples targeted for screening, namely, flugestone acetate, norges- were initially pooled by district and analysed by group tomet, delmadinone acetate, altrenogest, chlormadinone (n = 21). One pool and three individual samples for which acetate, melengestrol acetate and MPA and their respective results were not validated by the laboratory’sinternalquality CCα. Data acquisition was based on a targeted approach control were discarded from the final analysis. Fifty samples using SRM for the acquisition of four specific signals for among the first 10 positive pools were tested individually. each analyte. MPA transitions were 387.3 > 285.3, 387.3 > 123.2, 387.3 > 97.1 and 387.3 > 327.3. MPA-d3 fi Laboratory methods transition was 390.3 > 288.3. Before quanti cation of the MPA in kidney fat samples, the chemical residue, when Sample preparation detected, was identified according to the 2002/657/EC Kidney fat samples were extracted, purified and analysed decision criteria (relative retention time and transition in LABERCA (Oniris, Nantes, France) under ISO relative intensities) (European Commission 2002). Matrix 17025:2005 standards. The reference steroids, MPA and calibration curves were used for MPA quantification (5 – −1 MPA-d3, were, respectively, purchased from Steraloids, points, range 0 50 µg kg ). Inc., Ltd (London, UK) and RIKILT (Wageningen, The Netherlands). Most of the reagents and solvents were of analytical grade quality and provided by VWR Results International (Pessac, France). The SPE (C18 and silica) Method performance

Downloaded by [Cirad-Dist Bib Lavalette] at 10:04 22 November 2013 columns were from UCT (Bristol, PA, USA). Target pro- As can be seen from the ion chromatograms shown in gestagen’s analytes were first extracted by using acceler- Figure 1, the specificity and sensitivity of the analytical ated solvent extraction on 4 g of filtered kidney fat. method was fit-for-purpose. The multi-dimensional MS tech- Organic solvent was evaporated to dryness before succes- nology used for this study is nowadays recognised as the sive purification steps on SPE on C18 (2 g/10 mL) and gold standard for target residue analysis (De Brabander et al. silica (1 g/6 mL) cartridges. After evaporation to dryness, 2007; Noppe et al. 2008; Le Bizec et al. 2009). The use of a the dry residue was reconstituted by 50 µL of H2O/ACN labelled internal standard (MPA-d3)providedanaccurate mixture before injection (10 µL). quantification, as the recovery was automatically taken into account.

Detection and identiﬁcation of gestagens Main observations Data were acquired on a triple quadripole (Agilent 6410) operating in the positive ESI mode coupled to an LC The screening of the 80 kidney fat samples did not reveal system (Agilent 1200). The method used was validated residues of progestagens other than MPA. Out of the 19 according to the 2002/657/EC decision (Antignac et al. communes where animals were raised, synthetic hormone 2002) and decision limit (CCα) was calculated at residues were detected in pork carcasses originating from 14 Food Additives & Contaminants: Part A 3

Figure 1. (colour online). SRM chromatograms for MPA and MPAd3 in MRPL spiked (left), blank (middle) and non-compliant (right) kidney fat sample. Note: MPA, medroxyprogesterone acetate; MRPL, minimum required performance limit.

communes (73.6%). Figure 2 shows the communes where according to Malagasy regulation, and compared also with the MPA-positive samples were detected: (a) Ambilobe; (b) published literature (Heaton et al. 1996). The wide distribution Antsohihy and Bealanana; (c) Tsaratanana; (d) observed in the Malagasy provinces, mainly in central and Ankadinondry-Sakay, Maroharona, Tsiroanomandidy northern regions, confirmed a non-controlled extension of this Fihaonana, Tsinjoarivo and Miandrarivo; (e) fraud and the seriousness of the issue. Imerintsiatosika; (f) Ambatolampy, Ambohibary and The multi-residue detection method showed that only Antsirabe; (g) Ambohimahasoa; and (h) Betroka. MPA was present in our samples, confirming the large- MPA-positive samples were detected in 10 out of 15 districts scale misuse of one medicinal product only, and excluded (66.7%) and in all eight surveyed regions except one (87.5%), other synthetic progestagen substances at this time. namely, Diana region in Northern Madagascar. Out of the 50 Previous studies reported the spread of the MPA contam- individual samples selected among the first 10 positive pools, ination through the food chain by the supply of 10 fat kidney samples (20%) were tested positive for MPA MPA-contaminated liquid feed to pig farms by the com- Downloaded by [Cirad-Dist Bib Lavalette] at 10:04 22 November 2013 contamination, according to the criteria described in the 2002/ pound feed industry (Tielen 2003). However, because 657/EC regulation (Table 1). animal feed supply in Malagasy pig farms has been com- Table 2 indicates the number of pools of samples monly based on crop and local raw materials, and little on found to be contaminated within different ranges of levels compound feed, the hypothesis of a MPA contamination of progestin MPA. Concentrations of MPA in pools are of pork by animal feed was discarded, especially because distributed between 0.5 µg kg−1 and 139 µg kg−1.For the MPA concentrations in specimens were high. individual samples, concentrations of progestin MPA ran- Investigations by Malagasy veterinary services found ged from 0.5 µg kg–1 (for the less concentrated sample) to that farmers and animal health workers purchased syringes 187 µg kg–1 (for the most concentrated one). of progestagens (Confiance™, equivalent to Depo- Provera® 150 mg mL–1 Sterile Suspension for Injection, Pfizer (New York, NY, USA)). These low-price progestins Discussion were made easily available in private local dispensaries Our results highlighted a significant contamination of pork and in basic health centres (public sector) thanks to the products by synthetic hormone residues, which was highly efforts of international development agencies in the frame- prevalent considering that such residues are not likely to occur work of the national family planning and reproductive 4 V. Porphyre et al.

Figure 2. Distribution map of localities belonging to the eight regions where the sampled female adult pigs originated, namely, Diana (a), Soﬁa (b), Betsiboka (c), Bongolava (d), Itasy (e), Vakinankaratra (f), Haute-Matsiatra (g) and Anosy (h). Localities are displayed in white where all animals were tested negative for MPA. They are displayed in black where at least one of the tested animals tested positive for MPA. Sampled districts are shaded (Madagascar, October 2012). Note: MPA, medroxyprogesterone acetate.

Table 2. Distribution of samples by pools (n = 20) according to better concern by developing agencies and the medical different ranges of progestin medroxyprogesterone acetate sector for possible side effects (Pﬁzer Canada Inc. 2011). detected in kidney fat collected from adult sows at slaughter- ﬁ houses, Madagascar (2012). In particular, signi cant risk factors for osteoporosis lar- gely exist in the Malagasy poor populations (e.g., alcohol

Downloaded by [Cirad-Dist Bib Lavalette] at 10:04 22 November 2013 Estimated hormone Number of pools Number of −1 abuse, chronic use of drugs, low body mass index or concentration (µg kg ) of samples individual samples eating disorder), and, consequently, involuntary but regu- 0726lar ingestion of MPA in food by women might increase 0–139bone disorders in human population (Matson et al. 1997; 1–20 4 21 Clark et al. 2004). MPA accumulation in organs over time 20–45 3 13 was not demonstrated, but menstrual irregularity and pos- – 45 100 2 7 sible immediate clinical effect (anaphylactic responses) of >100 1 1 MPA residues in food should be investigated. Finally, epidemiological surveys about reproductive and bone disorders in humanpopulation, especially in female teenagers, health programmes throughout the country (Ofﬁce of should be urgently promoted. Inspector General 2011). Without any control, farmers Because ofﬁcial routine inspection of pig carcasses at then had the opportunity to use MPA in pigs easily. abattoirs by veterinary services is ineffective to rapidly Consequently, Malagasy Veterinary and Public Health detect both hormones and antimicrobial drug residues, Services should henceforth advocate for a better control of surveillance programmes using rapid tests should be pro- human drugs in the pharmaceutical retailing system and a moted to better estimate the risk for consumers. However, Food Additives & Contaminants: Part A 5

the analytical strategy needs to be re-thought within the This first study confirmedthepresenceofsynthetic framework of subsequent risk management programmes hormones in pork kidney fat produced in Madagascar with the competent authority according to local con- whatever the production area was, and MPA is the only straints, for example, the financial aspects, the availability progestagen that has been isolated. It showed the need of MS techniques and technical capacity in laboratories. for a national monitoring and information system dedi- LC-MS-MS was the method of choice in our situation for cated to the livestock sector and animal products to better detecting multi-residues at low concentrations under a risk protect Malagasy consumers. assessment approach. Although no previous information was available about either the field situation or the Acknowledgements expected concentration in the analysed matrix, the The authors gratefully thank the Service de Coopération et LC-MS-MS technique was suitable for a large screening d’Action Culturelle from the French Embassy in Madagascar, of multiple residues, including both progestins and andro- the Regional Council of La Reunion (French overseas territory), gens. But, because only one component, that is, MPA, was the European Regional development Funds and the French gov- detected in Madagascar with relatively high concentra- ernment for their financial support through the QualiREG tions, ELISA methods, whose detection limits are of the research network in Indian Ocean (www.qualireg.org). Thanks −1 also to Mrs. Ravaomanana Fleurette, Mrs. Rabibisoa Lalao order of 0.5 µg kg for the screening of MPA in kidney Francine and Mr. Rabenarivahiny René for their technical assis- fat, may be appropriate for veterinary inspection and could tance and for collecting biological samples in abattoirs. be put in place in Madagascar without a large investment (Chifang et al. 2006; Peng et al. 2008). GC-MS/MS References (Smets et al. 1997) or high performance thin layer chromatography (HPTLC) methods may also be an alternative Antignac J-P, Le Bizec B, Monteau F, Andre F. 2002. Validation of analytical methods based on mass spectro- for MPA monitoring (De Brabander & Van Hoof 1990). In metric detection according to the “2002/657/EC” European addition, for on-farm control, MPA, because of its lipo- decision: guideline and application. Anal Chim Acta. philic properties, could not be detected in urine but on the 483:325–334. contrary very well in faeces (Hamoir et al. 1998; Impens Braden BB, Talboom JS, Crain ID, Simard AR, Lukas RJ, Prokai et al. 2002, 2003). HPTLC and ELISA could be used to L, Scheldrup MR, Bowman BL, Bimonte-Nelson HA. 2010. Medroxyprogesterone acetate impairs memory and alters the detect MPA in faeces, but LC-MS-MS could be used as a GABAergic system in aged surgically menopausal rats. reference method (gold standard). However, faeces consti- Neurobiol Lear Mem. 93:444–453. tute a complex matrix to analyse and need many steps for Chifang P, Chuanlai X, Zhengyu J, Xiaogang C, Liying W. 2006. purification. Fat tissues sampled by subcutaneous biopsies Determination of anabolic steroid residues (medroxyproges- of superficial fat would be more appropriate to investigate terone acetate) in pork by ELISA and comparison with liquid chromatography tandem mass spectrometry. J Food Sci. 71: MPA contamination in live animals. C44–C50. However, African swine fever and other epizootic Clark M, Sowers M, Nichols S, Levy B. 2004. Bone mineral outbreaks have dramatically affected the Malagasy pig density changes over two years in first-time users of depot sector since 1998 (Ravaomanana et al. 2011)andledto medroxyprogesterone acetate. Fertil Steril. 82:1580–1586. the collapse of large-scale commercial farms. Resource- Clark MK, Dillon JS, Sowers M, Nichols S. 2005. Weight, fat mass, and central distribution of fat increase when women poor smallholder pig farms are now predominant use depot-medroxyprogesterone acetate for contraception. (Randriamparany et al. 2005; Ravaomanana et al. Int J Obes Relat Metab Disord. 29:1252–1258. 2011). Biosecurity measures in these small-scale pig Commission of the European Communities. 2002. Commission Downloaded by [Cirad-Dist Bib Lavalette] at 10:04 22 November 2013 farms are poorly implemented (Costard et al. 2009; Decision, 2002/657/EC implementing Council Directive Madec et al. 2010), and pig farmers have poor knowl- 96/23/EC concerning the performance of analytical methods and the interpretation of results (Text with EEA relevance) edge about veterinary drugs (Rakotoharinome et al. (notified under document number C(2002) 3044). Off J Eur 2013). Thus, public awareness campaigns are urgently Commun. L221:8–36. needed to alert pig farmers and professionals about the Commission of the European Communities. 2003. Commission negative consequences for public health of such misuse Decision, 2003/181/EC amending Decision 2002/657/EC as of synthetic hormones in animal. They should also clarify regards the setting of minimum required performance limits fi (MRPLs) for certain residues in food of animal origin (Text the content of of cial regulations, that is, prohibition of with EEA relevance) (notified under document number C synthetic hormones in livestock farming, sanctions and (2003) 764). Off J Eur Commun. L71:17–18. control measures to implement. Veterinary universities Costard S, Porphyre V, Messad S, Rakotondrahanta S, Vidon H, and training programmes for technicians in animal pro- Roger F, Pfeiffer DU. 2009. Multivariate analysis of man- duction should also disseminate relevant up-to-date agement and biosecurity practices in smallholder pig farms in Madagascar. Prev Vet Med. 92:199–209. information to students and young professionals to Courtheyn D, Le Bizec B, Brambilla G, De Brabander HF, improve their basic knowledge about veterinary drug Cobbaert E, Van de Wiele M, Vercammen J, De Wasch K. use in livestock production and about drugs residues in 2002. Recent developments in the use and abuse of growth animal products. promoters. Anal Chim Acta. 473:71–82. 6 V. Porphyre et al.

De Brabander HF, Le Bizec B, Pinel G, Antignac J-P, Verheyden growth hormone on muscle fibre characteristics, metabolic K, Mortier V, Courtheyn D, Noppe H. 2007. Past, present potential and meat quality. Meat Sci. 39:375–385. and future of mass spectrometry in the analysis of residues of Peng C-F, Chen Y-W, Chen W, Xu C-L, Kim J-M, Jin Z-Y. 2008. banned substances in meat-producing animals. J Mass Development of a sensitive heterologous ELISA method for Spectrom. 42:983–998. analysis of acetylgestagen residues in animal fat. Food De Brabander HF, Van Hoof J. 1990. HPTLC analysis of residues of Chem. 109:647–653. anabolics in meat and kidney fat. J Planar Chromatogr. 3:236-242. Pfizer Canada Inc. 2011. Product monograph – DEPO- Hamoir T, Pottie G, Courtheyn D, De Brabander H, Delahaut P, PROVERA et de DEPO-PROVERA-SC (acétate de Leyssens L. 1998. Comparison of purification procedures for médroxyprogestérone) [Internet]. [cited 2013 Feb 20]. the isolation and detection of anabolic residues in faeces Available from: http://www.pfizer.ca/fr/our_products/pro- using gas chromatography mass spectrometry. 3rd ducts/monograph/181 International Symposium on Hormone and Veterinary Drug Rakotoharinome M, Andriamanivo H, Blaga R, Perret C, Lacour Residue Analysis; 1998 Jun 2–5. Oud St Jan (Belgium): SA, Grasset-Chevillot A, Mace P, Thomas M, Villena I, Analyst; p. 2621–2624. Aubert D, et al. 2012. Toxoplasmosis and trichinellosis: an Heaton KL, Smith GC, Sofos JN, Aaronson MJ, Jones DK. epidemiological survey of pig population in Madagascar. 1996. Analysis of pork products for chemical residues. J Cluj-Napoca: XI European Multicolloquium of Parasitology. Muscle Foods. 7:213–224. Rakotoharinome M, Randriamparany T, Pognon D, Chane Ming Impens S, Courtheyn D, De Wasch K, De Brabander HF. 2003. J, Idoumbin JP, Cardinale E, Porphyre V. 2013. Prevalence Faster analysis of anabolic steroids in kidney fat by down- of antimicrobial residues in pork meat in Madagascar. Trop scaling the sample size and using gas chromatography-tan- Anim Health Prod. DOI: 10.1007/s11250-013-0445-9. dem mass spectrometry. 4th International Symposium on Randriamparany T, Grenier A, Tourette I, Maharavo Hormone and Veterinary Drug Residue Analysis; 2002 Jun Rahantamalala CY, Rousset D, Lancelot R. 2005. Situation 4. Antwerp, Belgium: Anal Chim Acta. 483:N1–N4, 1–432. épidémiologique de la peste porcine africaine dans la région Impens S, De Wasch K, Cornelis M, De Brabander HF. 2002. du Lac Alaotra et conséquences possibles pour l‘organisation Analysis on residues of estrogens, gestagens and androgens de la lutte et de la surveillance. Rev Elev Méd Vét Pays in kidney fat and meat with gas chromatography-tandem Trop. 58:15–20. mass spectrometry. J Chromatogr A. 970:235–247. Ravaomanana J, Jori F, Vial L, Pérez-Sánchez R, Blanco E, Ishida Y, Heersche JNM. 2002. Pharmacologic doses of medrox- Michaud V, Roger F. 2011. Assessment of interactions yprogesterone may cause bone loss through glucocorticoid between African swine fever virus, bushpigs activity: an hypothesis. Osteoporos Int. 13:601–605. (Potamochoerus larvatus), Ornithodoros ticks and domestic Le Bizec B, Pinel G, Antignac J-P. 2009. Options for veterinary pigs in north-western Madagascar. Transboundary Emerg drug analysis using mass spectrometry. J Chromatogr A. Dis. 58:247–254. 1216:8016–8034. Smets F, DeBrabander HF, Pottie G. 1997. Enhancing the speci- Madec F, Hurnik D, Porphyre V, Cardinale E. 2010. Good ficity of residue analysis for anabolics using HPLC clean up. practices for biosecurity in the pig sector – issues and Arch Lebensm Hyg. 48:30–34. options in developing and transition countries. Rome: FAO. Temmam S, Besnard L, Andriamandimby SF, Foray C, Malagasy Ministry of Agriculture. 2006. Malagasy regulation Rasamoelina-Andriamanivo H, Heraud JM, Cardinale E, relative to Livestock Production in Madagascar. 2006/030, Dellagi K, Pavio N, Pascalis H, et al. 2013. High prevalence Article 67. Antananarivo: Official Bulletin. of hepatitis E in humans and pigs and evidence of genotype- Matson S, Henderson K, McGrath G. 1997. Physical findings 3 virus in swine, Madagascar. Am J Trop Med Hyg. and symptoms of depot medroxyprogesterone acetate use in 88:329–338. adolescent females. J Pediatr Adolesc Gynecol. 10:18–23. Tielen MJM. 2003. The Medroxy Progesteron Acetate (MPA) – Meyer HHD. 2001. Biochemistry and physiology of anabolic case in Europe: an example of the weakest links in the hormones used for improvement of meat production. Quality Assurance System of the Feed Industry to produce APMIS. 109:1–8. Wholesome Food for All International Congress in Animal Noppe H, Le Bizec B, Verheyden K, De Brabander HF. 2008. Hygiene; Feb. 2003; Mexico: ISAH; p. 51–58. Novel analytical methods for the determination of steroid Unusan N. 2008. Determination of clenbuterol in UHT milk in Downloaded by [Cirad-Dist Bib Lavalette] at 10:04 22 November 2013 hormones in edible matrices. Anal Chim Acta. 611:1–16. Turkey. Int J Food Sci Technol. 43:617–619. Office of Inspector General. 2011. Audit of USAID/ van Leengoed L, Kluivers M, Herbes R, Langendijk P, Stephany Madagascar’s family planning and reproductive health activ- R, van den Berg M, Seinen W, Grinwis G, van der Lugt J, ities. Pretoria: USAID. Meulders F, et al. 2002. The weakest link: medroxyproges- Oksbjerg N, Petersen JS, Sórensen MT, Henckel P, Agergaard N, terone acetate in pig feed. Tijdschr Diergeneeskd. Bejerholm C, Erlandsen E. 1995. The influence of porcine 127:516–519.