Efek Minuman mengandung Bakteri Baik terhadap Profil Mikrobiota Fekal dan Penyerapan Kalsium Tikus Bunting serta Panjang Femur Bayi Tikus
Date
2021Author
Nurwati, Yuni
Hardinsyah, Hardinsyah
Marliyati, Sri Anna
Fahrudin, Mokhamad
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Seribu hari pertama kehidupan (1000 HPK) merupakan masa pertumbuhan bayi yang dimulai sejak masa konsepsi sampai usia dua tahun. Periode ini disebut periode emas karena berlangsung pertumbuhan otak dan organ – organ internal yang sangat pesat. Apabila kekurangan gizi pada periode tersebut, maka terjadi sebagian kerusakan atau kegagalan pertumbuhan dan tidak dapat diperbaiki pada periode berikutnya (Kemenkes 2014). Salah satu msalah gizi yang sering dihadapi pada 1000 HPK, yaitu kurang energi kronik (KEK) ibu hamil. Di Indonesia, prevalensi ibu hamil yang mengalami KEK sebesar 17,3% pada tahun 2018 (Kemenkes 2018). KEK selama kehamilan dapat menyebabkan kegagalan pertumbuhan janin, berat lahir bayi rendah (BBLR), cacat lahir, stunting, dan kematian bayi (Wu et al. 2012). Stunting merupakan kondisi gagal tumbuh yang ditandai dengan tinggi badan seorang anak di bawah -2SD (standar deviasi) atau sampai -3SD di usia dan jenis kelamin yang sama berdasarkan Child Growth Standards of The World Health Organization tahun 2006. Stunting dapat terjadi mulai dari janin dan terlihat pada saat anak berusia dua tahun (MCA 2014). Prevalensi balita stunting di Indonesia sebesar 30,8% (Kemenkes 2018) dan jika tidak dikendalikan, maka beresiko mengalami obesitas di masa mendatang (Black et al. 2013).
Kalsium, salah satu mineral makro, sangat dibutuhkan selama kehamilan. Kecukupan kalsium selama kehamilan baik untuk kesehatan tulang ibu dan janin, serta dapat mengurangi kejadian hipertensi selama kehamilan (Camargo et al. 2013). Di Kamerun dan Thailand, ibu hamil yang mengalami defisiensi kalsium sebesar 94,6% dan 55% berturut-turut. Sementara itu, di Kabupaten Jember, Jawa Timur, Indonesia, sebanyak 81,2% ibu hamil tidak dapat memenuhi kebutuhan kalsium yang seharusnya didapatkan dari pangan harian (Purnasari et al. 2016).
Probiotik, bakteri yang dapat memberikan manfaat baik untuk kesehatan, dapat menjadi alternatif intervensi yang dapat diterapkan untuk membantu memperbaiki metabolisme kalsium dan outcome kehamilan. Raveschot et al. (2020) menunjukkan bahwa probiotik strain Lactobacillus casei, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus plantarum, Lactobacillus fermentum, dan Lactobacillus helveticus dapat membantu penyerapan kalsium secara in vitro. Penelitian observasional yang mengkaji diet selama kehamilan menunjukkan bahwa konsumsi dairy products >3 porsi/hari berhubungan positif terhadap pertumbuhan dan panjang tulang femur (Chang et al. 2003).
Penelitian-penelitian mengenai bakteri baik atau probiotik sudah banyak dilakukan, namun sejauh ini belum ada penelitian pada ibu hamil dengan status gizi kurang. Oleh karena itu, penelitian ini bertujuan untuk mengkaji pengaruh pemberian minuman mengandung bakteri baik Lactobacillus casei shirota strain (LcS), Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus paracasei, Lactobacillus rhamnosus, dan Streptococcus thermophillus pada tikus bunting kurang gizi, dengan tujuan khusus mempelajari pengaruhnya terhadap (1) perbaikan mikroflora usus tikus bunting kurang gizi, (2) bioavailabilitas Ca tikus bunting kurang gizi, (3) kadar kalsium serum tikus bunting kurang gizi, (4) penambahan berat badan tikus bunting kurang gizi, (5) berat lahir, panjang lahir, dan panjang femur bayi tikus, dan (6) kadar kalsium femur bayi tikus. Penelitian ini merupakan penelitian eksperimental dengan Rancangan Acak Lengkap (RAL). Ethical Clearance diperoleh dari Komisi Etik Hewan, Fakultas Kedokteran Hewan, IPB University dengan nomor 025/KEH/SKE/XII/2020. Sebanyak 24 ekor tikus betina Sprague Dawley usia 7 – 8 minggu dengan berat 150 – 250 gram dibedakan menjadi satu kelompok kontrol negatif (status gizi normal, A0) dan tiga kelompok kurang gizi. Kelompok kurang gizi terdiri dari kontrol positif (A1), kelompok perlakuan LcS (A2), dan kelompok perlakuan minuman mengandung empat strain bakteri baik (A3). Pemberian intervensi dilakukan selama 19 hari. Sebelum dan setelah intervensi, tikus diukur berat badan, kadar Ca serum, feses, urin, total BAL feses, dan total bakteri coliform feses.
Hasil sidik ragam menunjukkan bahwa induksi kurang gizi tidak berpengaruh nyata terhadap rata – rata jumlah bakteri asam laktat (P>0,05). Sementara itu, hasil sidik ragam menunjukkan bakwa induksi kurang gizi berpengaruh nyata terhadap total bakteri koliform (P<0,05). Kelompok A2 memiliki bakteri koliform nyata lebih tinggi dibandingkan A0. Setelah diberikan intervensi, total bakteri koliform kelompok A2 mengalami penurunan yang nyata. Pemberian minuman mengandung bakteri baik juga berpengaruh pada bioavailabilitas kalsium. Kelompok A2 dan A3 nyata mengalami perbaikan bioavailabilitas kalsium dibanding A0 (P<0,05). Kalsium serum antar kelompok tidak berbeda nyata, namun kadar kalsium serum berada pada rentang normal di akhir kebuntingan. Hasil sidik ragam menunjukkan bahwa pemberian minuman mengandung bakteri baik berpengaruh nyata terhadap penambahan berat badan tikus selama kebuntingan. Kelompok A2 memiliki penambahan berat badan nyata paling kecil diantara kelompok lainnya. Berat lahir paling besar ditemukan pada bayi tikus kelompok A3 (5,18 ± 0,88 g) dan nyata lebih besar dibanding kontrol (A0). Panjang lahir dan panjang femur tertinggi juga ditemukan pada kelompok A3 (A3 = 4,39 ± 0,47cm and 0,29 ± 0,05 cm) namun tidak berbeda nyata dengan A0 dan A1. Hasil sidik ragam menunjukkan bahwa kadar kalsium nyata paling besar ditemukan pada kelompok A2.
Secara keseluruhan, pemberian minuman mengandung bakteri baik berpengaruh nyata terhadap perbaikan total bakteri koliform, bioavailabilitas kalsium, penambahan berat badan, dan outcome kehamilan. Oleh karena itu, pemberian minuman mengandung bakteri baik LcS maupun minuman dengan empat strain bakteri lainnya berpotensi sebagai alternatif intervensi untuk memperbaiki outcome kehamilan sehingga dapat membantu menurunkan masalah stunting pada balita. Saran untuk penelitian selanjutnya, perlu dilakukan penelitian yang mencampurkan kelima bakteri tersebut untuk membantu memperbaiki outcome kehamilan pada kondisi kurang gizi. Pemberian dosis bakteri juga diperhatikan supaya terlihat efek yang nyata pada outcome kelahiran. Selain itu, adanya potensi bakteri dalam memproduksi vitamin tertentu dan membantu penyerapan kalsium dapat menjadi suatu kajian tersendiri khususnya pada orang – orang dengan gangguan penyerapan kalsium ataupun pada kelompok vegetarian yang mengalami defisiensi vitamin B12.
Kata kunci : bioavailabilitas, kalsium, panjang femur, profil mikrobiota, tikus The first thousand days of life is a growth period of an infant starting from conception to the two years of age. It called “Golden Period” because of the rapid growth of the brain and internal organs. If there is a nutritional deficiency within this period, then some of the damage or growth disorders can not be repaired in the next period (Kemenkes 2014). One of many nutritional problems often encountered within 1000 HPK is a chronic energy deficiency (CED) in pregnant women. In Indonesia, the prevalence of CED in pregnant women was 17,3% (Kemenkes 2018). CED during pregnancy can cause fetal growth, low birth weight (LBW), birth defects, stunting, even infant death (Wu et al. 2012). Children are defined stunted if their height-for-age is more than two standard deviations below the Child Growth Standards of The World Health Organization 2006. Stunting occurs when the fetus is still in the womb and will be seen when the child is two years of age (MCA 2014). One of the risk factors for stunting is low body mass index in early pregnancy because it can interfere with the linear growth process of a baby (Pusparini et al. 2016). The prevalence of stunting in children under five years old in Indonesia was 30,8% (Kemenkes 2018) and if these prevalence not controlled, it can cause rapid weight gain after two years of age thus these children are at risk of developing obesity in the future (Black et al. 2013).
Calcium as one of the macro minerals is needed during pregnancy. The adequacy of calcium intake during pregnancy is good for maternal and fetal bone health which also able to reduce the hypertension incidence during pregnancy (Camargo et al. 2013). In Cameroon and Thailand, the deficiency of calcium in pregnant women were 94,6% and 55% respectively. Purnasari et al. (2016) showed that 81,2% pregnant women in Jember Regency, East Java, Indonesia, could not meet the calcium requirement which should had been obtained from their daily food.
Probiotics or lactic acid bacteria are able to provide many health benefits, could be use as one of the alternative interventions to improve calcium metabolism and pregnancy outcome. Raveschot et al. (2020) showed that Lactobacillus casei, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus plantarum, Lactobacillus fermentum, and Lactobacillus helveticus could improve the absorption of calcium in vitro. Observational studies examining diet during pregnancy have shown that consumption of dairy products >3 servings/day positively associated with growth and length of the femur (Chang et al. 2003).
Many studies on probiotics or good bacteria have been conducted, but so far there has been no research on pregnant women with CED. Therefore, this study aims to examine the effects of administration drink containing good bacteria Lactobacillus casei shirota strain (LcS), Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus paracasei, Lactobacillus rhamnosus, and Streptococcus thermophillus in undernourished pregnant rats. The specific objectives if this study are examine the effects of drink containing good bacteria on (1) the improvement of gut microflora in undernourished pregnant rats, (2) bioavailability of calcium in undernourished pregnant rats, (3) calcium serum in undernourished pregnant rats, (4) increment of body weight in undernourisged pregnant rats, (5) birth weight, birth length, and femur length of the offspring, and 6) calcium femur of the offspring. The design of the current study was experimental. All the studies were conducted according to the approval of Animal Ethics Commision, Faculty of Veterinary Medicine, IPB University No. 025/KEH/SKE/XII/2020. A total of 24 Sprague Dawley rats (7 – 8 weeks, 150 – 250 g) were assigned to the one negative control group (with a normal nutritional status, A0) and three undernourished groups. The undernourished groups were assigned as follows: a positive control group (A1), the treatment of drink containing LcS (A2), and the treatment containing four bacterias group (A3). Administration of drink containing good bacteria started in early pregnancy until 19 days of pregnancy. Before and after intervention, the data such as body weight, calcium in serum, calcium in faeces, calcium in urin, total lactic acid bacteria (LAB), and total coliform bacteria were measured.
Results in this study showed that induction of undernutrition did not affect the total LAB (P>0,05). Meanwhile, the induction of undernutrition significant affect the total coliform bacteria (P<0,05). The total coliform bacteria in A2 group was significantly higer from control group (A0). Administration of drink containing good bacteria significantly decreased the total of coliform bacteria in A2 group. Besides, it also affect the calcium bioavailability. The A2 and A3 groups have significantly increment of calcium bioavailability (P<0,05). For the serum calcium, there was no differences in serum calcium among groups, but the serum calcium of all groups was in the normal range at the end of pregnancy. Administration of drink containing good bacteria also significantly affect the increment of body weight during pregnancy in rats. The A2 group had the lowest increment of body weight. For the birth weight, the A3 group had the highest birth weight (5,18 ± 0,88 g) and significantly different from control (A0). The highest birth length (4,39 ± 0,47cm) and femur length (0,29 ± 0,05 cm ) also found in A3 groups but not significantly different from A0 and A1. The highest calcium femur found in A2 group.
Overall, administration of drink containing good bacteria significantly affect the total coliform bacteria, calcium bioavailability, increment of body weight during pregnancy in rats, and pregnancy outcome. Thus, administration of drink containing LcS or other four bacterias has the potential as alternative intervention to improve pregnancy outcome so that it can help to reduce stunting in toddlers. Based on this research, it is necessary to conduct further research relating to the development of product which mix those good bacteria to improve pregnancy outcome in undernourished conditions. The dosage of good bacteria that will be administered to the rats also need to be considered so that the effect could be seen clearly. In addition, the potential of bacteria to produce certain vitamins and help calcium absorption need to be conducted in further research particularly in people with impaired calcium absorption or deficiency of vitamin B12 as in vegetarian group.
Keywords: bioavailability, calsium, femur length, microbiota profile, rats
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