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Articles by A.D. Talpur
Total Records ( 7 ) for A.D. Talpur
  A.D. Talpur , A.J. Memon , M.I. Khan , M. Ikhwanuddin , M.M. Danish Daniel and A.B. Abol-Munafi
  Vibrio harveyi, Vibrio parahaemolyticus, Pseudoalteromonas piscicida, Staphylococcus epidermidis and Micrococcus luteus were isolated from the gut of blue swimming crab, Portunus pelagicus captured from Strait of Tebrau Johor Malaysia and studied for pathogenicity against the Zoea-1 (Z1 stage) of P. pelagicus. Pathogenic isolates V. harveyi and P. piscicida resulted in 100% mortality at 106 cfu mL-1 and 105 cfu mL-1 after 24 h and 72 h post dose. Conversely, V. parahaemolyticus produced 100% deaths at inoculation 106 cfu mL-1 after 72 h post dose. Cumulative mortality was observed rising with the increase in dose potency of pathogens. S. epidermidis and M. luteus detected with feeble pathogenic characteristics. The LD50 of V. harveyi was 1.2x103 cfu ML-1 (24 h), V. parahaemolyticus was 9.6x105 cfu mL-1 (72 h), P. piscicida was 9.8x103 cfu mL-1 (24 h) and S. epidermidis was 9.8x105 cfu mL-1 (72 h). The mean differences among various pathogenic doses were statistically significant (p<0.05). Susceptibility tests of total 662 isolates were under taken including V. harveyi (n = 180), V. parahaemolyticus (n = 180) and P. piscicida (n = 119), isolates showed mixed trend as multiple resistance and sensitive to antimicrobial agents tested while S. epidermidis (n = 88) and M. Luteus (n = 95) were sensitive to all antibiotics tested. V. harveyi, V. parahaemolyticus and P. piscicida did not show 100% resistance to any of the antibiotics tested. From the results of 14 antibiotics tested, we observed that the highest frequency of single drug resistance in V. harveyi was Streptomycin (89.44%) and sensitive to chloramphenicol (70.55%). Similarly, the highest frequency of single-drug resistance in V. parahaemolyticus was to kanamycin (92.78%) and sensitive to chloramphenicol (93.33%) and P. piscicida was to penicillin (80.67+19.33% intermediate but no sensitive) and sensitive to gentamicin (98.32%). Infections caused by antibiotic resistant pathogens have serious consequences and therapeutic use of tested antibiotic is questionable in larviculture of P. pelagicus.
  A.J. Memon , A.D. Talpur , M.I. Khan , M.O. Fariddudin , J. Safiah , A.B. Abol-Munafi and M. Ikhwanuddin
  The objectives of this study were to determine the effects of different cryoprotectants on sperm viability and optimization of spermatophore cryopreservation protocol for durable storage of Banana shrimp (Penaeus merguiensis). Spermatophore suspended for 15 min in Calcium-Free saline (Ca-F saline), used CPA MgCl2 and with concentration (15%), thawing temperature was 27°C. Use 15 min equilibration in room temperature (25°C) overall. Exposure and cooling rate selected as 25, 20, 16, 4, 2, -4, -20, -80, -150°C/10 min. Examination of sperm viability used a modified eosin-nigrosin staining technique. The smallest reductions in apparent sperm viability occurred with MgCl2, however freezing protocol was developed using Ca-F saline containing 15% MgCl2. Spermatophores were cryopreserved using above exposure/cooling rate and -196°C in liquid nitrogen up to 180 days. Mean sperm viability for fresh (93.8±1.3%) and cryopreserved spermatophore held for 24 h and 60 days was 83.5±0.6 and 61±1.2 did not differ (p>0.05), however that for spermatophore stored in liquid nitrogen between 90 and 180 days were lower (p<0.05) and varied from 55.4±0.3-16.4±1.2. Spermatophores earlier held in liquid nitrogen for 60 and 90 days. However, storage beyond 90 days caused a significant decline (p<0.05) in sperm viability. Spermatophores kept for 120 and 150 days had viabilities of 48.9±0.9 and 32.4±0.9%, respectively. Cryopreserved spermatophore stored in liquid nitrogen from 150-180 days had low viabilities (<35%). Mean fertilization rate of P. merguiensis females artificially inseminated with cryopreserved spermatophore that had been stored in liquid nitrogen for 7-30 days and for 60-90 days were 73.9±1.5-66.7±3.1 and 67.3±3-64.1±2.1%, respectively whereas that of fresh spermatophore was 88.2±1.5%. Hatching rates of eggs fertilized with cryopreserved spermatophore kept for 7-30 days and for 60-90 days were 77.6±2.5-72.7±3.5 and 81.5±12.1-62.5±1.5 which were not different (p>0.05) from those of the control group 76.2±13.5%, respectively. In conclusion, Cryopreserved spermatophore held in liquid nitrogen l<90 days revealed high sperm viability although, for longer periods, sperm viability declined at 180 days.
  A.D. Talpur , A.J. Memon , M.I. Khan , M. Ikhwanuddin , M.M. Danish Daniel and A.B. Abol-Munafi
  Blue swimming crab, Portunus pelagicus has not anchored the roots in aquaculture due to non availability of commercial seed production. Letdown of seed production is owing to microbial infections. To combat with microbes, study was aimed to isolate and screen probiotics from the gut of female crab for larviculture. Based on characteristics of inhibitory activity against pathogenic V. harveyi, V. parahaemolyticus and P. piscicida, bile, acid, salt tolerances and survival in sea water, isolates were identified as L. plantarum, L. salivarius, L. rhamnosus W. confusa and W. cibaria and evaluated for probiotics. A new model small scale in vivo validation was developed for conformity of the isolates as probiotics for P. pelagicus larviculture. The LAB isolates were administrated as water additive at concentrations 102, 104 and 106 cfu mL-1 for one day and five days in vivo validation experiments and positive control was inoculated with same concentrations of V. harveyi while negative control employed with larvae and no inoculation. Highest larval survival achieved at concentration 106 cfu mL-1 and L. plantarum, L. salivarius and L. rhamnosus did show significant larval survival. W. confusa and W. cibaria did not demonstrate as probiotics. L. plantarum showed highest survival 49.45±4.80% and 54.44±6.74% in both inoculations, respectively and no survival was observed in five days+ve control. Water quality degradation was not evident but improvement in pH was noticed. Based on results of small-scale in vivo test three LAB probiotics, L. plantarum, L. salivarius and L. rhamnosus were selected for larviculture of P. pelagicus.
  A.D. Talpur , A.J. Memon , M.I. Khan , M. Ikhwanuddin , M.M. Danish Daniel and A.B. Abol-Munafi
  The cause of mass mortality of Portunus pelagicus larvae reared in a hatchery system was investigated. The gut content of 180 female crabs and egg specimen of 24 female were studied for pathogenic microbes. The gut of female crabs were harboring fish pathogenic bacteria includes Staphylococcus epidermidis, Vibrio harveyi, Vibrio parahaemolyticus, Micrococcus luteus and Pseudoalteromonas piscicida and eggs were found associated with fish pathogens include Vibrio harveyi, Micrococcus luteus and Pseudoalteromonas piscicida. A causative transmitting pathogen V. harveyi through the feces of adult female crab and responsible for heavy mortality during larval rearing was determined by examining samples associated with the gut, hatching tanks, eggs, larvae rearing tanks, live and dead larvae of P. pelagicus. All isolates were identified by 16S rRNA gene sequencing. Vibrio harveyi was the major pathogen associated with all sources brought under study. Larvae were found to harbor a higher number of bacteria than larvae rearing tank. Experimental challenges with various doses indicated that the V. harveyi isolates were highly pathogenic. Doses 105 cfu mL-1 produced upto 96.67% mortality and 106 cfu mL-1 resulted in 100% mortality within 24 h post challenge. The differences among various doses of pathogen were statistically significant (p<0.05). The presence of these pathogens in P. pelagicus beyond the consequence for larval rearing is of epidemiological and health significance to humans.
  A.J. Memon , M. Ikhwanuddin , A.D. Talpur , M.I. Khan , M.O. Fariddudin , J. Safiah and A.B. Abol-Munafi
  The study was conducted to every hour assess the sperm viability of banana shrimp (Penaeus merguiensis) collected from Kedah water, West Malaysia (5°39'N; 100°19'E). Evaluations were done on three ways: group A: whole fresh specimens maintained at 2°C prior to extraction, group B: whole spermatophore maintained at 2°C and C: sperm suspension maintained at 2°C. Spermatophores counts were determined by sperm suspension using modified eosin-nigrosin staining method. Percentage of mean sperm viability for the fresh sample in group A at time zero was 93.96±3.74%, B was 98.8±0.7% and C was 99.02±0.7%. In group A, viability of the sperm considerably decreased after the first 60 min was 76.4±5.96% but in group B and C was gradually decreased at 91.4±0.9 and 97.6±1.2%. These were 67.9±5.13 and 62.5±5.1% in group A, 82.3±1.4% and 75.9±2.2% in group B and in group C was 93.3±1.9 and 88.2±3.1%, respectively after 1st-2nd h times elapsed. At 7th h in all groups the viability decreased significantly to <60% (p<0.05). Following, mean sperm viability were considerably decreased to group A was 50.7±4.79 and 37±6.52%, group B was 51.2±1.6 and 31.6±1.4% and group C was 56.2±1.9 and 41.5±2.3%, respectively after 7th and 9th h. However, for spermatophores in group A after 7th h fertilization and hatching rate was 44.3 and 64.4% in group B was 61.9 and 67.7% and group C was 42.9 and 61.3%, respectively at rates comparable to control 88.2 and 76.2%, respectively. There was no significant relationship was observed between biomass of the spermatophore to the body weight of the shrimp (p>0.05). The present study also revealed that specimens, spermatophore or sperm suspension maintained at 2°C could be utilized for fishery management through artificial insemination process till 7th h.
  A.J. Memon , N. Hidayati , A.D. Talpur , M.I. Khan , M.O. Fariddudin , J. Safiah , M. Ikhwanuddin and A.B. Abol-Munafi
  In aquaculture industry, spermatophore transfer technique such as artificial insemination in particular with reference to banana shrimp P. merguiensis is challenging. The aim of this study is to examine a novel technique for artificial insemination in P. merguiensis using SHDAI (Shrimp Holder Device for Artificial Insemination). In order to transfer the spermatophore properly into thelycum, an appropriate shrimp holder with continuous aeration system has been developed. During the process of manual spermatophore transfer (artificial insemination) also, a protective device to keep the female P. merguiensis shrimps alive and under minimum stress is necessary. The artificial insemination process carried out using SHDAI showed no signs of stress and/or mortality of the broodstock. During the experiments, female shrimps were 100% alive and active. Altogether, 78 female shrimps were tested of which 63 successfully accepted the spermatophore by using SHDAI. The accepted spermatophore percentage was significantly higher and achieved as 80.76%. Accepted spermatophore mass in the frozen, control and mean of both were recorded as 48.36±10.45, 45.0±15.28 and 46.68±2.38% which indicated no significant differences (p<0.05) However, frozen sperm at -196°C LN up to 90 days was 64.1±4.3% which indicated no significant differences between the SHDAI and control (p<0.05). Whereas frozen sperm at 196°C up to 90 days in LN was 62.5±2.9% there was no significant difference between the SHDAI and control (p<0.05). In the present study, overall quality of sperm in terms of the fertilization rate and hatching rate were almost similar between inseminated (using SHDAI) and control (natural mating) broodstock.
  A.J. Memon , A.D. Talpur , M.I. Khan , M.O. Fariddudin , J. Safiah , A.B. Abol-Munafi and M. Ikhwanuddin
  The spermatophore morphology of the P. merguiensis from Kedah, Malaysia is described. About 10 cryopreserved groups as 6, 12 and 24 h, 7, 30, 60, 90, 120, 150 and 180 days and fresh as control was examined from each of three replication were evaluated for sperm gross morphology evaluations. A fully mature male’s broodstock of P. merguiensis was taken with fresh spermatophores and evaluated for sperm morphologically. Cryopreserved spermatophore after the thawing 27°C/2 min (fresh and frozen) individually transferred into glass homogenizer (High speed variable speed reversible, Glas-col, Terre Havte In USA) with 200 μL of Ca-F saline. Fixation, dehydration by series of alcohol, Critical Point Dry (CPD) and mount specimens on to stubs using or carbon dots as well as using Auto Fine Coater and Sputter Coater moreover scanning by Model JEOL 6360LA scanning electron microscope. The cryopreserved spermatophore shows similarities with those of fresh, there were no significant differences (p>0.05) between the freshly spermatophore and spermatophorestored up to 90 days at -196°C liquid nitrogen.
 
 
 
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