Conversion of energy and protein rich food waste as an alternative feed ingredient in broiler feed formulation.

A series of experiments were performed to find out the impact of food waste on growth attributes and performance of broilers in a tropical climate. Two hundred and fifty-one-day-old broiler chicks were randomly separated into 5 groups, where each group comprised 50 animals. The broilers were fed with five different dietary treatments. Treatment 1 (T1), the diet consisted of food waste ingredients such as sprat heads, fish offal (protein), scraped coconut, and swill cooked rice as energy supplements; dietary treatment II (T2) diet was formulated with protein rich food waste; treatment III (T3) diet formulated with energy-rich food waste; treatment IV (T4) without any food waste materials, but a diet formulated with commercially available feed ingredients; and treatment V (T5), a 100% commercially-available broiler diet. Total feed intake per week and total weight gain were significantly (p < 0.05) higher in the commercial diet (T5) contrary to the formulated diets. The highest feed conversion ratio (FCR) was recorded in T3. The average dressing percentage was not significantly different (p > 0.05) in T1, T3, and T5. Average DM % in litter and DM % in feces were higher in T5, but average nitrogen % in droppings were lower in T4 and T5 compared to other diets. The study shows the potential application of food waste as an alternative feed in the broiler industry and its abundance and easy collection makes it a promising feeding regime in urban and suburban areas.

Anemia in Sri Lanka: A Literature Review.

Anemia is a global health problem. This paper reviews literature on the prevalence of anemia in Sri Lanka. We searched EBSCO (Elton Bryson Stephens Company), Cochrane Library, and Medline for articles on prevalence and molecular basis of anemia in Sri Lanka from January 2000 to May 2021. Forty articles were selected. Most of the studies were on prevalence of anemia among children and pregnant women. All the studies had restricted themselves to assess the contributing factors for anemia in limited age categories. Most articles had attempted to determine the overall prevalence of anemia and the contribution of iron deficiency to it. There were only a few studies on prevalence and molecular basis of hemoglobinopathies and even fewer on the prevalence of anemia of chronic disease. None of the studies had attempted to assess the national prevalence of red cell membranopathies and enzymopathies. The published data on prevalence of anemia in Sri Lanka are incomplete. This review emphasizes the value of a much broader survey on anemia covering all age categories including the elderly and conducting a national survey including anemia of chronic disease and on red cell membranopathies and enzymopathies in Sri Lanka.

A comparative study on fatty acid profile in selected vessels of coronary artery bypass graft (CABG).

Atherosclerosis is one of the leading non-communicable diseases in Sri Lanka. Analysis of fatty acid composition in blood vessels is important in understanding the development of atherosclerosis. Here, analyses of fatty acid profiles in major arteries which are commonly used in Coronary Artery Bypass Graft surgery (CABG) were subjected to investigation. Patients (n = 27) undergoing elective CABG were enrolled in the study. A small biopsy segment of the saphenous vein (SV), radial artery (RA), and left internal mammary artery (LIMA) of patients was obtained during the surgery. The fatty acid (FA) profile of tissue samples was analyzed using Gas Chromatography-Mass Spectroscopy (GCMS). Among the different arteries tested, palmitic acid and stearic acid were the predominant fatty acids. As far as monounsaturated FA (MUFA) are concerned, oleic acid was found to be the most abundant MUFA in vessels. The FA profile of LIMA samples had a higher SFA percentage and lower unsaturated FA percentage compared to other vessels. Furthermore, the vessel samples of RA indicated the highest percentage of pro-inflammatory ω -6 polyunsaturated fatty acids (PUFA) as well as a higher percentage ratio between ω -6: ω -3 PUFA. The fatty acid composition and ω -6: ω -3 PUFA ratio suggests that LIMA graft is preferred for CABG over RA and SV.

Proximate Composition, Physicochemical, Functional, and Antioxidant Properties of Flours from Selected Cassava (Manihot esculenta Crantz) Varieties.

Cassava flour has a high potential to contribute as a raw material in the food industry. This study was aimed at characterizing flours from Sri Lankan cassava varieties with a view to explore the potential in food applications. Flours prepared from five cassava varieties, namely, Kirikawadi, MU51, Swarna, Shani, and Suranimala, were analyzed for proximate composition and physicochemical, functional, and antioxidant properties using standard methods. Flours from tested cassava varieties contained <1% crude fat and <2% crude protein. Flour from MU51 contained the highest amount of HCN (48.05 mg/kg) while flour from Suranimala contained the lowest (4.85 mg/kg). Total starch and amylose contents of flours were significantly lower (p < 0.05) than those of commercial wheat flour. Flour from Suranimala contained approximately similar amylopectin content as commercial wheat flour. Water absorption capacity, oil absorption capacity, water solubility index, swelling power, emulsion activity, and emulsion stability of flours from five cassava varieties were significantly higher (p < 0.05) than those of commercial wheat flour. Swarna was identified as the richest source of phenolic compounds (4.44 mmol GAE/100 g dry weight) among the five varieties. Results showed the promising application potential of flours from these five cassava varieties in different food applications such as weaning foods, bakery foods, and edible films.

Anther Culture in Coconut (Cocos nucifera L.).

Doubled haploids have a high impact on the improvement of heterozygous crops through hybridization. Anther culture is a doubled haploid technique for producing homozygous lines. In coconut, a tree species reported to be recalcitrant for tissue culture, a successful doubled haploid protocol was established through anther culture. All the factors affecting androgenesis induction have been optimized. In this chapter, a stepwise protocol, from doubled haploid induction including palm selection, anther isolation, pretreatment, and culture initiation, up to plant regeneration and thereafter acclimatization of the regenerated plants, is described. Furthermore, the protocol for testing the anther-derived plants for the ploidy level is also presented.

Efficacy and Safety of the Two Ayurveda Drug Regimens in Uterine Fibroids: A Randomized Single-Blind Clinical Trial.

This study aims to assess the efficacy and safety of two Ayurveda drug regimens for the treatment of uterine fibroids (UF) in a randomized single-blind clinical trial. 120 participants with UF (volume ≥ 2 cm3) were randomly allocated at a 1 : 1 : 1 ratio to 2 experimental groups and the control group. The 12-week intervention period was followed by 12-week follow-up. The primary efficacy endpoint was the change of the largest UF volume. The secondary efficacy endpoints were assessed by the pictorial bleeding assessment score (PBAC), UF symptoms, and quality of life score. The safety endpoints were changed in hepatic and renal safety parameters and patients experiencing adverse effects. Significant decrease was observed in the volume of UF in the arm II but not in arm I, while a significant increase was observed in the volume of the largest UF in the control group at 12th week. The PBAC score remained stable in all groups. Further mean value of the intervention arms symptom severity subscale (SSS) was significantly reduced compared to the control arm. Health-related quality of life (HRQL) value improved in 12th week of both experimental arms compared to baseline. Control arm HRQL value was reduced compared to baseline. The volume of the largest UF and both SSS and HRQL values remained stable within the follow-up period in the tested arms. The findings of this study demonstrated the safety and efficacy of selected two Ayurveda drug regimens in reducing the volume of UF and related symptoms and improving quality of life.

A study of morphological and morphometric variations in Sri Lankan femoral diaphysial nutrient foramina / Estudio de las variaciones morfológicas y morfométricas de los forámenes nutricios diafisarios de fémur en Sri Lanka

SUMMARY: Blood supply to the diaphysis of long bones is majorly through nutrient artery, which enters the bone via nutrient foramen. The present study aims to identify morphological and morphometric variations of nutrient foramina of the femur, which is the largest long bone of the body extending from the pelvis to knee. A sample of 81 Sri Lankan adult femur were analyzed. The mean length of the femur analyzed was 436.93 mm and mean foramen index was 43.52. The majority of the bones had a single nutrient foramen. The predominant location of the nutrient foramen was on the posterior aspect of the bone in the middle third of the shaft according to the study. Majority of foramina were directed distally. The indices on nutrient foramen are important for procedures such as bone grafts and tumor resections, in managing trauma, orthopedic procedures and radiological interpretations.

RESUMEN: El suministro de sangre a la diáfisis de los huesos largos se realiza principalmente a través de la arteria nutricia, que ingresa al hueso a través del foramen nutricio diafisario. El presente estudio tiene como objetivo identificar las variaciones morfológicas y morfométricas del foramen nutricio del fémur, el hueso largo más grande del cuerpo que se extiende desde la pelvis hasta la rodilla. Se analizó una muestra de 81 fémures adultos de individuos de Sri Lanka. La longitud media de los fémures analizados fue de 436,93 mm y el índice de foramen medio fue de 43,52. La mayoría de los huesos tenían un solo foramen nutricio. Según el estudio, la ubicación predominante del foramen nutricio estaba en la cara posterior del hueso en el tercio medio del eje. La mayoría de los forámenes estaban dirigidos distalmente. Los índices sobre el foramen nutricio son importantes para procedimientos tales como injertos óseos y resecciones tumorales, en el manejo de traumatismos, procedimientos ortopédicos e interpretaciones radiológicas.

How well does Malaysia achieve value for money in public sector purchasing of medicines? Evidence from medicines procurement prices from 2010 to 2014.

BACKGROUND: Malaysia's public healthcare sector provides a greater volume of medicines at lower overall cost compared to the private sector, indicating its importance in providing access to medicines for Malaysians. However, the Ministry of Health (MOH) has concerns about the continuous increase in the public sector medicines budget, and achieving efficiencies in medicines procurement is an important goal. The objectives of this study were to assess the overall trend in public sector pharmaceutical procurement efficiency from 2010 to 2014, and determine if the three different ways in which MOH procures medicines influence efficiency. METHODS: We matched medicines from the public sector procurement report by medicine formulation to medicines with a Management Sciences for Health (MSH) International Reference Price (IRP) for each year. Price ratios were calculated, and utilizing the information on quantity and expenditure for each product, summary measures of procurement efficiency were reported as quantity- and expenditure-weighted average price ratios (WAPRs) for each year. Utilizing MOH procurement data to obtain information on procurement type, a multiple regression analysis, controlling for factors that can influence prices, assessed whether procured efficiency (relative to IRPs) differed by MOH procurement type. RESULTS: Malaysia's public sector purchased medicines at two to three times the IRP throughout the study period. However, procurement prices were relatively stable in terms of WAPRs each year (2.2 and 3.2 in 2010 to 1.9 and 2.9 in 2014 for quantity and expenditure WAPRs, respectively). Procurement efficiency did not vary between the three different methods of MOH procurement. Procurement efficiency of both imported originators and imported generics were significantly lower (P < 0.001 and P < 0.01) than local generic products, and medicine source and category influenced the procurement efficiency of each MOH procurement mechanism. CONCLUSION: The design of different medicines procurement mechanisms, along with the balance between ensuring competitive procurement prices and adhering to national industry and procurement policies, have not been able to achieve lower public sector medicines procurement prices (relative to IRP). Introducing pooled procurement options along with continuous monitoring of procurement efficiency and exploring ways to improve price competition among local and foreign suppliers is recommended.

Development of Pasta Products with Nonconventional Ingredients and Their Effect on Selected Quality Characteristics: A Brief Overview.

Pasta is a widely consumed food in all over the world. Coarse semolina obtained from durum wheat and water are the main ingredients of conventional pasta products. The amount of gluten and quality level of durum wheat, are two important factors for the superiority of finished pasta. Market price of durum wheat is higher than the common wheat and it contributes no more than 5% of the world wheat production. Thus, to come across the challenge of emerging pasta consumption, new field of research that is dealing with the incorporation of nonconventional ingredients to the conventional formula of pasta has initiated. The compositions of raw materials which are used for pasta preparation directly affect the physical, chemical, and textural properties of the product. Therefore, incorporation of nonconventional ingredients can lead to a contradictory effect of pasta quality. This review will focus on the various types of nonconventional ingredients that are being incorporated in pasta products and their effect on the quality attributes of different pasta products.

Inter-relationship of serum leptin levels with selected anthropometric parameters among a non-diabetic population: a cross-sectional study.

PURPOSE: Association between serum leptin levels and anthropometric parameter is well established in western countries according to the specific WHO cut-off values assigned for those populations, whereas it is not clearly defined for Asians especially with respect to gender. Thus, the objectives of this study were to determine the relationship of serum leptin levels with body mass index (BMI), waist circumference (WC), waist to hip ratio (WHR) to identify the variations of serum leptin levels with gender and to evaluate the serum leptin levels in risk and non-risk groups based on their anthropometric values. SUBJECTS/METHODS: A cross-sectional study was carried out among 226 apparently healthy subjects (non-diabetics, age 20-70 years). Height, weight, WC, hip circumference (HC) and mid arm circumference (MAC) were measured. BMI, WHR, waist to height ratio (WHtR) were calculated. Fasting blood samples were collected. Serum leptin levels were measured using human leptin ELISA kits. RESULTS: Majority of the participants were females (59.3%). Serum leptin levels were significantly higher in females (24.8 ± 17.1 ng/mL) compared to males (9.3 ± 7.9 ng/mL). Significant positive correlations (P < 0.05) were observed between serum leptin levels and all anthropometric parameters except height in both genders. The risk groups according to BMI, WC and WHR in females were hyperleptinaemic and had significantly (P < 0.05) higher serum leptin levels than the non-risk groups. CONCLUSIONS: Linear trend was observed for serum leptin levels with weight, BMI, WC, HC, WHR, MAC and WHtR in both genders. Though the serum leptin levels were higher among risk groups, according to WHR, WC and BMI, the hyperleptinaemia was observed only among females. LEVEL OF EVIDENCE: A descriptive cross-sectional study, Level V.

Detection of Rater Errors on Cognitive Instruments in a Clinical Trial Setting.

OBJECTIVES: This study examines errors committed by raters in a clinical trial of a memory enhancement compound. BACKGROUND: Findings of clinical trials are directly dependent on the quality of the data obtained but there is little literature on rates or nature of rater errors on cognitive instruments in a multi-site setting. DESIGN: Double-blind placebo-controlled study. SETTING: 21 clinical sites in North America. PARTICIPANTS: Two hundred seventy-five participants. MEASUREMENTS: MMSE, WMS-R Logical Memory I and II, WMS-R Verbal Paired Associates I, WASi Vocabulary, WASi Matrix Reasoning, GDS and MAC-Q. RESULTS: The WMS-R Logical Memory I and II and WASi Vocabulary tests were found to have the greatest number of scoring errors. Few substantive errors were detected on source document review of the MMSE, GDS, MAC-Q and WMS-R Verbal Paired Associates I. Some additional administration and scoring issues were identified during feedback sessions with the raters. CONCLUSIONS: Cognitive measures used in clinical trials are prone to errors which can be detected with proper monitoring. Some instruments are particularly prone to inter-rater variably and should therefore be targets for focused training and ongoing monitoring. Areas in need of further investigation to help inform and optimize quality of clinical trial data are discussed.

Antidiabetic Compounds in Syzygium cumini Decoction and Ready to Serve Herbal Drink.

Herbal beverages with desirable sensory attributes are an ideal way to offer consumers with phytochemicals having specific health promoting functionalities. Syzygium cumini bark decoction is used in treating diabetes mellitus in Ayurveda medicine. This work attempted to prove the presence of antidiabetic compounds in the S. cumini decoction and the ready to serve (RTS) herbal drink developed using the decoction. Activity guided fractionation of the decoction of the S. cumini was carried out by sequential extraction with organic solvents of different polarities. Thin Layer Chromatography (TLC) with authentic compounds and HPLC were performed for identification and confirmation of the compounds in the decoction and the RTS herbal drink. Presence of gallic and ellagic acids in the decoction and RTS herbal drink was proven and confirmed with HPLC. The LC UV-VIS spectra of the two compounds were identical with the corresponding spectra of the library. Further, gallic acid and umbelliferone were determined as the active compounds in the decoction by TLC and were confirmed by cochromatography with authentic compounds. The present investigation confirmed the presence of gallic acid, ellagic acid, and umbelliferone which are proven to have antidiabetic activities in the decoction and the RTS herbal drink prepared with the decoction.

Effects of two litter amendments on air NH3 levels in broiler closed-houses.

OBJECTIVE: High NH3 emissions from poultry houses are reported to have negative impacts on health, welfare and safety of birds and humans, and on the environment. Objective of the present study was to determine the effects of two litter amendments on the NH3 levels in broiler closed houses under hot-humid conditions. METHODS: Giving a completely randomize design, nine closed houses, each housed 32,500 birds on paddy husk litter, were randomly allocated into two treatment (Mizuho; a bacterial culture mix and Rydall OE; an enzymatic biocatalyst) and control groups. NH3 levels were determined thrice a day (0600, 1200, and 1800 h), at three heights from the litter surface (30, 90, and 150 cm), at 20 predetermined locations of a house, from day 1 to 41. RESULTS: Rydall significantly reduced the NH3 level compared to control and Mizuho. NH3 levels at 30 cm were significantly higher than that of 90 and 150 cm. The NH3 levels at 30 cm height were higher than 25 ppm level from day 9, 11, and 13 in Mizuho, control, and Rydall groups, respectively to day 41. NH3 levels at 150 cm height were higher than maximum threshold limit of 50 ppm for human exposure from day 12, 14, and 15 in Mizuho, control, and Rydall groups, respectively to day 33. Being significantly different among each other, the NH3 level was highest and lowest at 0600 and 1800 h. Litter amendments had no significant effects on growth performance. Rydall significantly increased the litter N content on day 24. CONCLUSION: It was concluded that the NH3 levels of closed house broiler production facilities under tropical condition are so high that both birds and workers are exposed to above recommended levels during many days of the growing period. Compared to microbial culture, the enzymatic biocatalyst was found to be more effective in reducing NH3 level.

Exercise-driven metabolic pathways in healthy cartilage.

OBJECTIVE: Exercise is vital for maintaining cartilage integrity in healthy joints. Here we examined the exercise-driven transcriptional regulation of genes in healthy rat articular cartilage to dissect the metabolic pathways responsible for the potential benefits of exercise. METHODS: Transcriptome-wide gene expression in the articular cartilage of healthy Sprague-Dawley female rats exercised daily (low intensity treadmill walking) for 2, 5, or 15 days was compared to that of non-exercised rats, using Affymetrix GeneChip arrays. Database for Annotation, Visualization and Integrated Discovery (DAVID) was used for Gene Ontology (GO)-term enrichment and Functional Annotation analysis of differentially expressed genes (DEGs). Kyoto Encyclopedia of Genes and Genome (KEGG) pathway mapper was used to identify the metabolic pathways regulated by exercise. RESULTS: Microarray analysis revealed that exercise-induced 644 DEGs in healthy articular cartilage. The DAVID bioinformatics tool demonstrated high prevalence of functional annotation clusters with greater enrichment scores and GO-terms associated with extracellular matrix (ECM) biosynthesis/remodeling and inflammation/immune response. The KEGG database revealed that exercise regulates 147 metabolic pathways representing molecular interaction networks for Metabolism, Genetic Information Processing, Environmental Information Processing, Cellular Processes, Organismal Systems, and Diseases. These pathways collectively supported the complex regulation of the beneficial effects of exercise on the cartilage. CONCLUSIONS: Overall, the findings highlight that exercise is a robust transcriptional regulator of a wide array of metabolic pathways in healthy cartilage. The major actions of exercise involve ECM biosynthesis/cartilage strengthening and attenuation of inflammatory pathways to provide prophylaxis against onset of arthritic diseases in healthy cartilage.

Dehydration and malaria augment the risk of developing chronic kidney disease in Sri Lanka.

Chronic kidney disease (CKD) of unknown etiology (CKDu) is a serious health issue in Sri Lanka. One-to-one age and sex-matched two sample comparative study was carried out in the Medawachchiya divisional secretariat area of the North Central Province (NCP) of Sri Lanka, by randomly selecting 100 CKDu patients and 100 age and sex-matched subjects from non-CKDu affected families from the same area. An interviewer-administered questionnaire was used for the collection of data pertaining to occupation, medical history and lifestyle. Data were analyzed using a conditional linear logistic model. Working for >6 h in the field per day, exposure to sun, drinking water only from well, consumption of <3 L of water per day, and having a history of malaria were found to be having significant (P < 0.05) likelihood toward the development of CKDu. Treatment of water prior to consumption had a significant protective effect against CKDu. Dehydration, history of malaria and drinking untreated well water from are likely contribute to the development of CKD of unknown etiology among the inhabitants of NCP, Sri Lanka.

Follistatin-like 3 is a mediator of exercise-driven bone formation and strengthening.

Exercise is vital for maintaining bone strength and architecture. Follistatin-like 3 (FSTL3), a member of follistatin family, is a mechanosensitive protein upregulated in response to exercise and is involved in regulating musculoskeletal health. Here, we investigated the potential role of FSTL3 in exercise-driven bone remodeling. Exercise-dependent regulation of bone structure and functions was compared in mice with global Fstl3 gene deletion (Fstl3-/-) and their age-matched Fstl3+/+ littermates. Mice were exercised by low-intensity treadmill walking. The mechanical properties and mineralization were determined by µCT, three-point bending test and sequential incorporation of calcein and alizarin complexone. ELISA, Western-blot analysis and qRT-PCR were used to analyze the regulation of FSTL3 and associated molecules in the serum specimens and tissues. Daily exercise significantly increased circulating FSTL3 levels in mice, rats and humans. Compared to age-matched littermates, Fstl3-/- mice exhibited significantly lower fracture tolerance, having greater stiffness, but lower strain at fracture and yield energy. Furthermore, increased levels of circulating FSTL3 in young mice paralleled greater strain at fracture compared to the lower levels of FSTL3 in older mice. More significantly, Fstl3-/- mice exhibited loss of mechanosensitivity and irresponsiveness to exercise-dependent bone formation as compared to their Fstl3+/+ littermates. In addition, FSTL3 gene deletion resulted in loss of exercise-dependent sclerostin regulation in osteocytes and osteoblasts, as compared to Fstl3+/+ osteocytes and osteoblasts, in vivo and in vitro. The data identify FSTL3 as a critical mediator of exercise-dependent bone formation and strengthening and point to its potential role in bone health and in musculoskeletal diseases.

Rare hemoglobin variants: Hb G-Szuhu (HBB: c.243C>G), Hb G-Coushatta (HBB: c.68A>C) and Hb Mizuho (HBB: c.206T>C) in Sri Lankan families.

In this short communication, we describe the clinical presentation of unusual hemoglobin (Hb), variants in three Sri Lankan cases under study for ß-thalassemia intermedia (ß-TI). We believe this is the first report on their occurrence in Sri Lanka as well as from the Indian subcontinent. During a molecular study performed on ß-TI patients, we identified three unusual Hb variants as Hb G-Szuhu (HBB: c.243C>G), Hb G-Coushatta (HBB: c.68A>C) and Hb Mizuho (HBB: c.206T>C) in three unrelated families. Hb G-Szuhu and Hb G-Coushatta were found in combination with the common ß-thalassemia (ß-thal) mutation, IVS-I-5 (G>C). Both probands had mild anemia with greatly reduced red cell indices and had non palpable livers and spleens, however, by ultrasound, both were observed to be enlarged. The final Hb variant, Hb Mizuho, was identified as a heterozygous mutation found in both proband and his mother. Both family members had severe anemia and were regularly transfused and had increased red cell parameters.

Reprogramming of cassava (Manihot esculenta) microspores towards sporophytic development.

Gametes have the unique potential to enter the sporophytic pathway, called androgenesis. The plants produced are usually haploid and recombinant due to the preceding meiosis and they can double their chromosome number to form doubled haploids, which are completely homozygous. Availability of the doubled haploids facilitates mapping the genes of agronomically important traits, shortening the time of the breeding process required to produce new hybrids and homozygous varieties, and saving the time and cost for inbreeding. This study aimed to test the feasibility of using isolated and in vitro cultured immature cassava (Manihot esculenta) microspores to reprogramme and initiate sporophytic development. Different culture media and different concentrations of two ion components (Cu(2+) and Fe(2+)) were tested in two genotypes of cassava. External structural changes, nuclear divisions and cellular changes during reprogramming were analysed by scanning electron microscopy, by staining with 4',6-diamidino-2-phenylindole, and through classical histology and transmission electron microscopy. In two cassava genotypes, different developmental stages of microspores were found to initiate sporophytic cell divisions, that is, with tetrads of TMS 60444 and with mid or late uni-nucleate microspores of SM 1219-9. In the modified NLN medium (NLNS), microspore enlargements were observed. The medium supplemented with either sodium ferrous ethylene-diamine-tetraacetic acid (NaFeEDTA) or CuSO4·5H2O induced sporophytic cell division in both genotypes. A low frequency of the reprogramming and the presence of non-responsive microspores among the responsive ones in tetrads were found to be related to the viability and exine formation of the microspores. The present study clearly demonstrated that reprogramming occurs much faster in isolated microspore culture than in anther culture. This paves the way for the development of an efficient technique for the production of homozygous lines in cassava. This is the first ever detailed report of microspore reprogramming at the tetrad stage and the first report of microspore embryogenesis induction in cassava with detailed evidence.