Haematological profile of common carp spawners of various breeds

The goal of this study was to determine the basic haematological profile of common carp (Cyprinus carpio L.) spawners of eight breeds reared under identical conditions and sampled in spring after overwintering. Significant differences were found among the breeds for haemoglobin level (Hb), haematocrit value (PCV), mean corpuscular volume (MCV) and mean corpuscular haemoglobin (MCH) values. The number of red blood cells (RBC), mean corpuscular haemoglobin concentration (MCHC), number of leukocytes (WBC) and leukocyte differential count (WBC DIFF) did not differ significantly among the respective breeds. The highest Hb, PCV and MCH values were found for Ropsha scaly carp (ROP) and Amur wild carp (AS) (Hb 109 ± 17 and 106 ± 15 g l^?1; PCV 0.40 ± 0.06 and 0.40 ± 0.06 l l^?1; MCH 69 ± 7 and 69 ± 9 pg, respectively). The AS specimens and breeds that originated [ROP, Ukraine scaly carp (US), and Northern mirror carp (M72)] showed significantly higher values of Hb (P < 0.01), PCV (P < 0.01), MCV (P < 0.05) and MCH (P < 0.05), compared to the other four breeds: Hungarian mirror carp (M2), Israeli mirror carp (Dor 70), South Bohemian mirror carp (BV) and Tata scaly carp (TAT). Males showed significantly higher PCV, Hb and RBC values within individual breeds. This study demonstrated that Amur wild carp and breeds originating from it had significantly higher values of erythrocyte profile in comparison with the other breeds studied.     

Haematological and serum biochemical reference values of snow trout,Schizothorax labiatus habiting in river Sindh of Indian Himalayan region

Haematological and serum biochemical parameters are a valuable tool in determining the wellbeing status of different fish species and response in close association to various biological and environmental factors, therefore the aim of the present study was to establish sex-wise reference values of haemato-biochemical parameters of S. labiatus. The haematological parameters such as haemoglobin (Hb) concentration, total erythrocyte count (RBC), total leucocyte count (WBC), packed cell volume (PCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), mean corpuscular volume (MCV) and serum biochemical parameters such as glucose, cholesterol, urea, total protein, albumin and globulin were analysed. The results revealed that the haematological parameters, i.e., Hb, RBC, WBC and PCV showed significant (P < 0.05) differences with respect to sex. Significantly (P < 0.05) higher values of Hb content (11.18 g dl⁻¹), RBC count (1.88 × 10⁶ mm⁻³) and PCV (36.15%) were noted in male fish than females throughout the study period, while significantly (P < 0.05) higher WBC count was noted in females (20.38 × 10³ mm⁻³) compared to males. However, no significant (P > 0.05) difference was observed in erythrocyte indices like MCH, MCHC and MCV between sexes. The serum biochemical parameters, i.e., glucose, cholesterol and urea, also showed significant (P < 0.05) difference between the sexes, but no significant (P > 0.05) difference was noticed in total protein, albumin and globulin. Significantly (P < 0.05) highest values of glucose (116.76 mg dl⁻¹) and urea (9.01 mg dl⁻¹) were recorded in male S. labiatus, while highest value of cholesterol (223.53 mg dl⁻¹) was noted in females. The information generated in the present study gives the basic reference values of haematological and serum biochemical parameters of S. labiatus which will be useful in monitoring the wellbeing status of fish populations. Moreover, the data will also be helpful to ascertain the occurrence of different clinical and subclinical diseases, which may in turn help to boost the overall production of this species.     

Age, sexual and seasonal differences of haematological values and antibody status to Chlamydophila sp. in feral and racing pigeons (Columba livia forma domestica) from an urban environment (Zagreb, Croatia)

The aim of this study was to determine the basic haematological parameters in feral and racing pigeons and to compare these parameters according to age, sex and season in healthy feral pigeons as well as between Chlamydophila-serologically positive and negative feral pigeons. Red blood cells (RBC), packed cell volume (PCV), haemoglobin (Hb), mean cell volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), white blood count (WBC), thrombocyte count and differential WBC, were determined in 366 pigeons (Columba livia forma domestica) captured in the City of Zagreb between 1999 and 2002. Of these, 232 feral (179 adult and 53 juvenile, 104 male and 75 female) and 57 racing pigeons (25 male and 32 female) were clinically healthy and bacteriologically and serologically negative, but 77 birds had antibody titres against Chlamydophila sp. Significantly lower values of RBC, PCV, Hb, MCH, WBC and thrombocyte (P<0.05) were observed in young compared to adult pigeons, while the differences in MCV and MCH were not significant between age classes. In differential WBC of young pigeons, a significantly higher percentage of heterophils, eosinophils, basophils and monocytes and a significantly smaller percentage of lymphocytes (P<0.01) was found than in adult pigeons. Significant sex-related differences were seen only in MCV values and in the percentage of lymphocytes (higher in females) and neutrophils (higher in males). PCV, Hb, MCV and MCH increased, while WBC decreased during wintertime (P<0.05). In differential WBC, percentage of heterophils was low in summer and autumn. At the same time, a higher percentage of basophils was found. Low numbers of monocytes were found in summer and low values of eosinophils in winter. In racing pigeons, values of eosinophils and basophils were significantly lower than in feral pigeons. Pigeons which had antibodies against Chlamydophila sp. possessed a higher percentage of monocytes and less lymphocytes than sero-negative animals, while WBC was significant lower than in sero-negative feral pigeons.     

Biological and molecular responses of Chironomus riparius (Diptera,Chironomidae) to herbicide 2,4-D (2,4-dichlorophenoxyacetic acid)

2,4-Dichlorophenoxyacetic acid (2,4-D) is an agricultural contaminant found in rural ground water. It remains to be determined whether neither 2,4-D poses environmental risks, nor is the mechanism of toxicity known at the molecular level. To evaluate the potential ecological risk of 2,4-D, we assessed the biological parameters including the survival rate, adult sex ratio of emerged adults, and mouthpart deformities in Chironomus riparius after long-term exposure to 2,4-D. The larvae were treated with 0.1, 1 or, 10 μg L^? 1 of 2,4-D for short- and long-term exposure periods. The sex ratio was changed in C. riparius exposed to only 10 μg L^? 1 of 2,4-D, whereas mouthpart deformities were observed as significantly higher in C. riparius exposed to 0.1 μg L^? 1 of 2,4-D. Survival rates were not significantly affected by 2,4-D. Furthermore, we evaluated the molecular and biochemical responses of biomarker genes such as gene expression of heat shock proteins (HSPs), ferritins and glutathione S-transferases (GSTs) in C. riparius exposed to 2,4-D for 24 h. The expressions of HSP70, HSP40, HSP90 and GST levels in C. riparius were significantly increased after exposure to a 10 μg L^? 1 concentration of 2,4-D, whereas ferritin heavy and light chain gene expressions were significantly increased at all concentrations of 2,4-D exposure. Finally, these results may provide an important contribution to our understanding of the toxicology of 2,4-D herbicide in C. riparius. Moreover, the 2,4-D-mediated gene expressions may be generated by 2,4-D is the causative effects on most probable cause of the observed alterations. These biological, molecular and morphological parameters and the measured parameters can be used to monitor 2,4-D toxicity in an aquatic environment.     

Some age related changes of adult rainbow trout, Salmo gairdneri Rich., peripheral erythrocytes separated by velocity sedimentation at unit gravity

Velocity sedimentation at unit gravity separated peripheral erythrocytes of adult rainbow trout according to age and size. The cells located at the top of the gradient contained numerous polyribosomes, few mitochondria and an essentially electronlucent nucleus. The cells at the bottom of the gradient no longer contained these cytoplasmic organelles and had an essentially electron-dense nucleus. The middle of the gradient contained cells with either very few polyribosomes or cells devoid of this organelle.
The length, width and mean corpuscular volume (MCV) of the average cell increased and the width to length ratio decreased progressively from the top of the gradient to the bottom.
No Statistical significance could be determined in the changes of the mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) although they appeared to follow theoretical projections.
The use of haematocrit, red cell count, haemoglobin concentration, MCV, MCH and MCHC in assessing erythropoietic activity and the size and haemoglobin content of the maturing cell is discussed.     

Increased whole blood manganese concentrations observed in children with iron deficiency anaemia

A prospective observational study was carried out at Alder Hey Children's Hospital, Liverpool, England, UK on children aged 1–6 years attending the pathology department for routine blood tests (n = 225). Whole blood manganese concentrations were measured plus the following markers of iron status; haemoglobin, MCV, MCH, RBC count, ferritin, transferrin saturation and soluble transferrin receptors. Multiple regression analysis was performed, with blood manganese as the dependent variable and factors of iron status, age and gender as independent variables. A strong relationship between blood manganese and iron deficiency was demonstrated (adjusted R² = 34.3%, p < 0.001) and the primary contributing factors to this relationship were haematological indices and soluble transferrin receptors. Subjects were categorised according to iron status using serum ferritin, transferrin saturation and haemoglobin indices. Children with iron deficiency anaemia had higher median blood manganese concentrations (16.4 μg/L, range 11.7–42.4, n = 20) than children with iron sufficiency (11 μg/L, range 5.9–20.9, n = 59, p < 0.001). This suggests that children with iron deficiency anaemia may be at risk from manganese toxicity (whole blood manganese >20 μg/L), and that this may lead to neurological problems. Treatment of iron deficiency in children is important both to improve iron status and to reduce the risk of manganese toxicity.     

Possible recovery from an acute envenomation in male rats with LD50 of Echis coloratus crude venom: I-A seven days hematological follow-up study

The effect of an acute LD₅₀ dose of Echis coloratus crude venom in male albino rats was tested on blood parameters: white blood cells (WBCs), red blood cells (RBCs), platelets count, hemoglobin, hematocrit, mean cell volume (MCV), mean cell hemoglobin (MCH) and mean cell hemoglobin concentration (MCHC), also serum glucose, total protein, triglycerides with alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and γ-glutamyl transferase (GGT) enzyme activities. The effect of the LD₅₀ dose was monitored over a period of seven days, with time intervals of 1, 3, 6, 12, 24, 72 h. All of the tested parameters show fluctuations with time and with tendency to regain normal control level after 12 h. At 12–24 h it seems to be crucial for the process of physiological recovery, in spite of the irreversible damage and tissue distraction. The process of physiological adaptation and recovery from the lethal destructive venom effect seems to stabilize after one week, leaving the animal alive with several biochemical altered metabolisms and disturbed physiological profile.     

Changes in the nucleotide triphosphate/haemoglobin and nucleotide triphosphate/red cell ratios of rainbow trout, Salmo gairdneri Richardson, subjected to prolonged starvation and bleeding

The nucleotide triphosphate/haemoglobin (NTP/Hb) and nucleotide triphosphate/red cell (NTP/cell) ratios of rainbow trout increased during prolonged starvation. A decline was noted in blood lactic acid concentration. Red cell count, haemoglobin concentration and haematocrit also declined. Changes in mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) were found not to be significant. The NTP/Hb and NTP/cell ratios of both fed and starved trout rose seven days following a 15% reduction in blood volume by cardiac puncture. A rise in whole blood NTP concentration was found only in the bleeding response of fed animals. No significant change was noted in blood lactic acid concentration. The decline in haematocrit was significant only in the starved group. In both groups, however, red cell count and blood haemoglobin concentration fell. MCV rose whereas MCHC declined in all bled animals. Changes in MCH were not significant in either group. Negative correlations were noted between red cell count and both the NTP/Hb and NTP/cell ratios and between haemoglobin concentration and the NTP/Hb ratio. Positive correlations were seen between the two ratios and between red cell count and haemoglobin concentration.     

Effect of high tannin grain sorghum on gastrointestinal parasite fecal egg counts in goats

The objective of three experiments was to determine the influence of high condensed tannin (CT) grain sorghum on gastrointestinal parasite fecal egg counts (FEC). Sixteen naturally infected Boer crossbred mixed-sex goats were used. Animals that were supplemented with grain daily were removed from pasture and placed in individual pens and fed treatment diets for 21 or 14 days (Experiment 3). Goats were allowed ad libitum access to water and diets containing high or low CT cracked grain sorghum. On day 0 and every 7 days thereafter, PCV, FEC and FAMACHA^® eyelid color scores (EYE; Experiment 1 only) were recorded. For Experiment 1, percentage of animals dewormed, PCV and EYE were not influenced by treatment and averaged 13.6 ± 4.5% per treatment period, 23.4 ± 0.8% and 3.2 ± 0.12, respectively, for all animals. The FEC increased after day 0 for control but not high tannin grain sorghum fed goats (treatment by day interaction, P < 0.02). As expected, EYE and PCV were negatively related (r = −0.45; P < 0.0005). For Experiment 2, there was no influence of diet on PCV or FEC, but there was an effect of day on FEC (P < 0.002) in which all goats had higher FEC on day 0 (1956 ± 219 eggs/g) than any other sampling day. For Experiment 3, there was no effect of feeding high CT grain sorghum on FEC (2992 ± 591 eggs/g) or PCV (25.1 ± 0.5%). In these studies, high CT grain sorghum did not consistently influence FEC and did not impact PCV or number of animals requiring chemical anthelmintic treatment.     

Classification of complete blood count and haemoglobin typing data by a C4.5 decision tree,a naïve Bayes classifier and a multilayer perceptron for thalassaemia screening

This article presents the classification of blood characteristics by a C4.5 decision tree, a naïve Bayes classifier and a multilayer perceptron for thalassaemia screening. The aim is to classify eighteen classes of thalassaemia abnormality, which have a high prevalence in Thailand, and one control class by inspecting data characterised by a complete blood count (CBC) and haemoglobin typing. Two indices namely a haemoglobin concentration (HB) and a mean corpuscular volume (MCV) are the chosen CBC attributes. On the other hand, known types of haemoglobin from six ranges of retention time identified via high performance liquid chromatography (HPLC) are the chosen haemoglobin typing attributes. The stratified 10-fold cross-validation results indicate that the best classification performance with average accuracy of 93.23% (standard deviation = 1.67%) and 92.60% (standard deviation = 1.75%) is achieved when the naïve Bayes classifier and the multilayer perceptron are respectively applied to samples which have been pre-processed by attribute discretisation. The results also suggest that the HB attribute is redundant. Moreover, the achieved classification performance is significantly higher than that obtained using only haemoglobin typing attributes as classifier inputs. Subsequently, the naïve Bayes classifier and the multilayer perceptron are applied to an additional data set in a clinical trial which respectively results in accuracy of 99.39% and 99.71%. These results suggest that a combination of CBC and haemoglobin typing analysis with a naïve Bayes classifier or a multilayer perceptron is highly suitable for automatic thalassaemia screening.     

Haematological studies on the prewintering and wintering frog, Rana esculenta

1. A study of the haematology of the frog Rana esculenta including erythrocyte count (RBC), haemoglobin content (Hb), haematocrit (HCT), mean cell volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC) and erythrocyte size as a function of prewinter and winter was made. 2. The RBC count and Hb were significantly higher in contrast to MCV and MCH values during prewinter in both sexes. 3. The surface area to volume ratio was higher in prewinter whereas the length to width ratio (eccentricity) of the cytosome and nucleus was significantly higher during winter in both sexes. 4. Sexual differences in the erythrocyte count, Hb content and the surface area to volume ratio were also observed. 5. The physiological significance of these observations are reported for Rana esculenta.     

Experimental studies on anaemia in sheep infected with Dictyocaulus filaria

The pathogenesis of Dictyocaulus filaria induced anaemia was experimentally investigated. Nineteen Dorset-Muzaffarnagri male lambs were divided into two groups of 13 and six. The lambs in the former group received a primary infection dose of 2000 D. filaria infective larvae, whereas animals in the latter group were kept as uninfected controls. The haematological alterations and faecal larval output were monitored weekly until day 220 post-infection. The infection caused an increase in erythrocyte sedimentation rate (ESR) and total leukocyte count (TLC). However, a decrease in packed cell volume (PCV), haemoglobin, total erythrocyte count (TEC) and blood pH was observed in acute infection. The shape and size of the erythrocytes, serum bilirubin level, mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) remained unaffected In carriers, except for TEC, the values of these parameters returned to near-normal levels. The analysis of the results suggests that the infected animals developed a normocytic normochromic anaemia which persisted during the later stage of infection.     

Effect in vivo of atrazine on haematology and O2 consumption in fish, Tilapia mossambica.

Studies were conducted on haematological constituents such as Red blood cells (RBC), White blood cells (WBC), Haemoglobin (Hb), Packed cell volume (PCV), Mean cell volume (MCV), Mean corpuscular haemoglobin (MCH), Mean corpuscular haemoglobin concentration (MCHC), Blood volume (BV), Blood water content (BWC) and Whole animal oxygen consumption (WAOC) in the fish exposed to sublethal concentration of atrazine. Significant changes were seen in the constituents of the blood and O2 consumption of fish suggesting the existence of respiratory distress in the fish as a consequence of atrazine toxicity.     

Effects of confinement stress and additional zinc treatment on some blood parameters in the dogfish Scyliorhinus canicula

Several blood parameters were studied in the dogfish Scyliorhinus canicula subjected to different confinement conditions (transport and maintenance) and zinc contamination. Confinement stress significantly decreased erythrocyte counts (RBCC), haematocrit (Ht), haemoglobin (Hb), leucocrit (Lt), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC). Mean corpuscular volume (MCV) did not change with this stress and plasma glucose concentration significantly increased. In fish subjected to confinement stress conditions, Zn treatment significantly decreased Hb, MCH and MCHC and plasma glucose concentration. Ht and MCV did not change and Lt and RBCC significantly increased. Results are discussed related to other stress effects and effects of zinc treatment on fish confined in non-stressful conditions.     

Performance of West African dwarf goats fed Guinea grass–Verano stylo mixture,N-fertilized and unfertilized Guinea grass

The supplementary values of Verano stylo in a mixed Guinea grass (Panicum maximum cv. Ntchisi)–Verano stylo (Stylosanthes hamata cv. Verano) diet from a sown grass–legume mixture and N fertilized grass were compared in West African dwarf (WAD) goats. Liveweight (LW) gain, feed intake, digestibility and N utilization were determined using 15 goats in two trials lasting for 98 days. Goats were fed Guinea grass–Verano stylo mixture (GSM), N-fertilized (NFG) and unfertilized grass (UFG). The goats were divided into three groups of five animals each and randomly allocated to the dietary treatments in a randomized complete block design. Total DM and OM intakes of the goats did not vary significantly among the forage diets and averaged 55.1 and 50.4 g kg⁻¹ W^0.75 per day, respectively. CP intake (g kg⁻¹ W^0.75 per day) was highest with NFG (5.6) followed by GSM (4.8) and the UFG (3.5). Total N excreted followed the same trend as the CP intake. There was no significant difference between N-retention of GSM and NFG (28.5 and 26.7%), but goats on UFG had a negative N balance (−9.16%). Animals on GSM had significantly higher liveweight gain (31.9 g per day) than those of NFG (25.1 g per day) and UFG (21.9 g per day) which also differed significantly. The digestibilities of total DM, OM, CP, NDF were higher with GSM than NFG or UFG. It is concluded that growing Verano stylo in mixture with Guinea grass is a better option for improving the feed quality of forage diets for goats than direct application of inorganic fertilizer at 200 kg N ha⁻¹ to the pure grass.     

Effects of 4-nonylphenol on blood cells of the African catfish Clarias gariepinus (Burchell, 1822)

In the present work, the destructive effects of the 4-nonylphenol on one of the most economically important Nile fishes, namely African catfish (Clarias gariepinus) were studied. Apoptosis, erythrocytes alterations, micronucleus test and blood parameters count were used as biological indicators to detect those effects. After exposure to sublethal concentrations of 4-nonylphenol (0, 0.05, 0.08 and 0.1 mg/l), apoptotic red blood cells with many malformations and micronucleated erythrocytes were recorded. Decrease in the blood parameters such as red blood cells (RBCs), hemoglobin (Hb), package cell volume (PCV), mean corpuscular hemoglobin concentration (MCHC), platelets, white blood cells (WBCs), lymphocytes, basophils, monocytes and increase in mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), neutrophils, eosinophils indicated the negative effects of 4-nonylphenol. It was concluded that, the 4-nonylphenol caused genotoxicity in erythrocytes with many malformations in shape and number indicated with other blood parameters.     

Microinjection of melanin concentrating hormone into the lateral preoptic area promotes non-REM sleep in the rat

The ventrolateral preoptic area (VLPO) has been recognized as one of the key structures responsible for the generation of non-REM (NREM) sleep. The melanin-concentrating hormone (MCH)-containing neurons, which are located in the lateral hypothalamus and incerto-hypothalamic area, project widely throughout the central nervous system and include projections to the VLPO. The MCH has been associated with the central regulation of feeding and energy homeostasis. In addition, recent findings strongly suggest that the MCHergic system promotes sleep. The aim of the present study was to determine if MCH generates sleep by regulating VLPO neuronal activity. To this purpose, we characterized the effect of unilateral and bilateral microinjections of MCH into the VLPO on sleep and wakefulness in the rat. Unilateral administration of MCH into the VLPO and adjacent dorsal preoptic area did not modify sleep. On the contrary, bilateral microinjections of MCH (100 ng) into these areas significantly increased light sleep (LS, 39.2 ± 4.8 vs. 21.6 ± 2.5 min, P < 0.05) and total NREM sleep (142.4 ± 23.2 vs. 86.5 ± 10.5 min, P < 0.05) compared to control (saline) microinjections. No effect was observed on REM sleep. We conclude that MCH administration into the VLPO and adjacent dorsal lateral preoptic area promotes the generation of NREM sleep.     

Temporal variations in haematological and biochemical indices of the Caspian kutum, <Emphasis Type="Italic">Rutilus frisii kutum</Emphasis>

The pattern of haematology and blood biochemistry variations of healthy adult female kutum Rutilus frisii kutum (Cyprinidae) was investigated in the three reproductive states: maturing (autumn), pre-spawning (winter), and spawning (spring). Red blood cells, haemoglobin, and haematocrit were highest in autumn, whereas the highest values for white blood cells, mean corpuscular volume (MCV), and mean corpuscular haemoglobin (MCH) were observed in winter. With the exception of MCV and mean corpuscular haemoglobin concentration (MCHC), fish size had no effect on haematological and biochemical parameters. MCV was higher in larger fish, whereas the reverse trend was found for MCHC. Decreased levels of plasma glucose, cholesterol, and triglyceride in the pre-spawning period, when female ovaries were developing, may have been due to the combination of reproductive effects and food shortage in winter. Total protein levels were lower during the maturing period than during the pre-spawning period. The results of this study suggest that temporal variations in haematological and biochemical parameters of blood are significant and should be considered when these parameters are used to assess fish health status.     

Erythrocytic parameters as indicators for differentiating between the pregnant and pseudopregnant bitches in Nigeria

The erythrocytic parameters during pregnancy and pseudopregnancy in bitches were studied and compared in 8 bitches aged 2 -3 years and weighing 10-12 kg. Blood samples were collected from the bitches before mating, during the three trimesters of pregnancy and the post partum period. The packed cell volume (PCV %), haemoglobin concentration (Hb gm/dl), red blood cell count(x106/μl) were determined using standard methods. The mean corpuscular volume (MCV) and mean corpuscular haemoglobin concentration (MCHC) were then calculated. Six of the bitches were pregnant and 2 were pseudopregnant. The results showed that in pregnant bitches, the PCV decreased significantly from the premating values of 51.37+0.94% to 34.00+8.04% during the third trimester of pregnancy (P.     

Indirect organogenesis from various explants of Hildegardia populifolia (Roxb.) Schott & Endl. – A threatened tree species from Eastern Ghats of Tamil Nadu,India

Hildegardia species are an important resource for fiber industry. This investigation was conducted to develop a plant regeneration protocol for Hildegardia populifolia (Roxb.) Schott & Endl. via indirect organogenesis Callus was obtained from leaf, internode and petiole explants, among these explants internode explant gave best result on MS medium supplemented with different concentrations of 2,4-Dichlorophenoxy acetic acid (2,4-D). The highest percentage (100%) of regeneration was obtained with benzyladenine (BA) (2.0 mg/l) + indole-3-acetic acid (IAA) (0.1 mg/l) + glutamine (25 mg/l) + thidiazuron (TDZ) (0.5 mg/l) from internode explants. Shootlets were highly rooted on MS medium supplemented with 3.0 mg/l indole-3-butyric acid (IBA). In vitro rooted seedlings were successfully acclimatized. This in vitro regeneration system will facilitate further development of reliable procedures for this genus.