Cryopreservation of silver barb Puntius gonionotus (Bleeker) spermatozoa: effect of extender composition,cryoprotective agents and freezing rate on their postthawing fertilization ability

The effects of extender composition, cryoprotectant concentration and freezing and thawing on the fertilization efficiency of cryopreserved spermatozoa of Puntius gonionotus were evaluated. Computer‐aided motility analysis of semen was conducted to check the suitability of spermatozoa for cryopreservation after mixing with different extenders and cryoprotective agents (CPAs). Extender‐4 with an osmolality 260 mOsmol kg⁻¹and pH 7.6 was used for the cryopreservation study. Among the CPAs, dimethyl sulphoxide (DMSO) was least toxic and more than 60% fertilization was achieved when used at 1.4 M at 0 °C for 10 and 30 min, whereas the toxicity of all CPAs to spermatozoa was evident when tested at 30 °C. Semen frozen at −16 °C min⁻¹ with 1.4 M DMSO showed 70% fertilization, which was significantly higher (P<0.05) than other freezing rates. Samples thawed at 35 °C water showed a fertilization rate comparable with that of fresh semen. Computer‐assisted semen analysis of fresh and frozen semen after thawing showed variations in different types of motility in spermatozoa and in their class. There was no significant difference in motility before or after cryopreservation; however, significant differences could be observed in the average path velocity (VAP), straight line velocity (VSL) and curve linear velocity (VCL). Semen of silver barb could be cryopreserved with extender‐4 by addition of 1.4 M DMSO to a final cryopreservation medium (MED 2) cooled at a rate of −16 °C min⁻¹, stored in liquid nitrogen (−196 °C) and utilized after thawing at 35±2 °C.     

Changes in ovary and testis and breeding fitness of the climbing perch,Anabas testudineus (Bloch, 1792), exposed to sub‐lethal concentrations of monocrotophos

Monocrotophos (MCP) is one of the organophosphate pesticides extensively used in agriculture and animal husbandry. The present study reports the effects of sub‐lethal concentrations of MCP on gonadosomatic index (GSI), gonadal histology and breeding fitness of the fish Anabas testudineus. The fish were exposed to three sub‐lethal concentrations of monocrotophos (T1: 3.5 mg/L, T2: 5.3 mg/L and T3: 10.6 mg/L) for 45 days with 12 fish (males and females in equal numbers) in each tank, and one group was kept as control (C) without any treatment. GSI decreased in all treatment groups in comparison with control. Significant decrease in fertilization and hatching rate was observed at all concentrations in comparison with control on 15, 30 and 45 days of monocrotophos exposure. The decreased vitellogenesis, disruption of follicular wall and oocyte atresia were observed in all the treated females at 30 and 45 days of exposure. Rupture of seminiferous tubules was noticed in all the treated males irrespective of exposure periods. The results thus indicated that monocrotophos could deleteriously impact the gonadal structure and function in A. testudineus, which could seriously impact the reproductive success of the animal.     

Effect of a Previous Washing Step with Tannic Acid on the Quality of Minced Horse Mackerel (Trachurus trachurus) During Frozen Storage

Fatty fish have been recognized as potential raw material for production of minced meat; however, they are prone to oxidation and further deterioration. In the present study, the effect of washing and antioxidant (tannic acid) treatment on the quality of minced meat of Trachurus trachurus (horse mackerel) during frozen storage was observed. Minced meat of Trachurus trachurus was divided into three lots (T0, T1, and T2). T1 was washed with cold water, T2 with cold water containing tannic acid (100 mg/kg), and T0 was not washed. All the lots were frozen at ?40°C and stored at ?20 ± 2°C for 125 days and were subjected to biochemical, microbiological, and sensory evaluation at regular intervals of 25 days. The antioxidant treatment with tannic acid at the dosage used was found effective in minimizing the rancidity problems of minced meat (T2), compared to T0 and T1. During the whole period of storage, samples from T2 showed good quality in terms of microbiological, biochemical, and sensory analysis compared to T1 and T0.     

Differential HSP90α expression in fish hepatocytes from polluted estuary during summer

The molecular chaperone, heat shock protein 90α (HSP90α), plays an important role in protein folding, degradation of denatured proteins and steroid activation. It is essential for the maintenance of cellular integrity and survival when induced in response to environmental, physical and chemical stresses. In the present investigation the effect of environmental stress on HSP90α expression was examined in grey mullet Mugil cephalus living in either a contaminated (Ennore) or uncontaminated (Kovalam) estuary over two seasons: Hepatocytes were isolated from grey mullet of both estuaries. Oxidative stress was determined along with HSP90α in these fish. Additionally, immunohistochemical changes were studied to confirm the HSP90α expression. Comparison of the results revealed enhanced hepatocyte oxidative stress and HSP90α expressions in fish from Ennore to a significant extent than fish from Kovalam. Also, the results showed significant seasonal variations with maximum expression observed during summer compared to the monsoon season. Overexpression of HSP90α in hepatocytes exposed to chronic environmental stress by pollutants may confer differential effects on cell survival by protecting against oxidative stress induced changes. The results also indicate that seasonal variations have significant effect on the HSP90α expression.     

Effect of dietary Curcuma longa on enzymatic and immunological profiles of rohu,Labeo rohita (Ham.), infected with Aeromonas hydrophila

The aim of this study was to evaluate the dietary dosages of turmeric that enhance immune response and disease resistance against the opportunistic pathogens Aeromonas hydrophila in Labeo rohita fingerlings. Hence, four different dosages of turmeric at 0.1, 0.5, 1.0 and 5.0 g kg^?1 feed were given to the fingerlings of L. rohita for 60 days at 4% body weight. At every 20‐day interval, different biochemical, haematological, enzymatic and immunological parameters of fish were evaluated. After 60 days, fish were challenged with A. hydrophila. The mortality (%) was recorded on the tenth day post challenge. Most of the immune parameters including lysozyme activity, superoxide anion production and serum bactericidal activity were significantly (P<0.05) higher on 60 days of feeding of 1.0 g of turmeric per kg of feed. Challenge study indicated 100% and 89% survivability in the group of fish fed with 5.0 and 1.0 g of turmeric per kg of feed respectively. Feeding of turmeric might have maintained long‐term protection in fish by elevating the nonspecific immune system such as Nitroblue tetrazolium (NBT), lysozyme and serum bactericidal activity. The result showed that turmeric at a dose of 1.0 g kg^?1 feed for 60 days provided the greatest protection to pathogen challenge.     

Effect of sub-lethal nitrite on selected haematological parameters in fingerling Catla catla (Hamilton)

A sub‐lethal nitrite toxicity trial was conducted using static conditions for a period of 96 h with fingerlings of Catla catla (21.4±3.6 g). Fingerlings were exposed to five concentrations of nitrite, that is, 1, 2, 4, 8 and 10.4 mg L^?1 and a nitrite‐free control to study changes in haematological parameters. Nitrite caused an increase in immature erythrocyte population (7–24%) in lower concentrations (0–4 mg L^?1) at 6 h while they were absent in higher concentrations. The total erythrocyte count was reduced at 6 h followed by an increase at 12 h with further reduction up to 96 h in all concentrations of nitrite. The 96‐h exposure resulted in 21.2–31.8% reduction in erythrocyte population in 1–10.4 mg L^?1 nitrite. The haemoglobin content decreased progressively with increasing nitrite concentrations as well as exposure periods. Total leukocyte count decreased initially at 6 h in all treatments followed by an increase after 12 h, signifying development of a protective response of the body to nitrite stress. Blood glucose decreased initially up to 24 h followed by an increase through 96 h. Serum protein level decreased continuously with increasing exposure period. The study revealed that exposure to nitrite caused changes in almost all the haematological parameters in the fingerlings depending on the concentration as well as exposure period. Nitrite being one of the important inorganic nutrients often recorded at higher levels in intensively cultured ponds, the present study highlights its adverse impact on fish and stressed the need for the management of this nutrient in culture ponds.     

Embryonic and larval developments of brackish water catfish,Mystus gulio (Hamilton and Buchanan, 1822) induced with human chorionic gonadotropin and consequent larval rearing

Mystus gulio, the long whiskers catfish, is a popular food fish and potential candidate species for aquaculture in Sundarban area of India and Bangladesh. Recently, catch of this species has declined due to overfishing and various ecological changes. In the present study, mature fish was induced to spawn in captivity through intramuscular injection of human chorionic gonadotropin at the doses of 10 IU/g to female and 5 IU/g to male. Photomicrographs of all developmental stages of live embryo and larvae were documented with the aid of a light microscope. Results demonstrated that morula, blastula, gastrula and neurula and organogenesis ended at 1:30‐, 3:00‐, 5:30‐, 7:30‐ and 17:15‐hour post‐spawning (hps) respectively. Heartbeat and muscular contraction of embryo commenced at 8:30 and 11.15 hps respectively. Hatching of embryo started after an incubation period of 17:30 hour at an ambient temperature of 29 ± 1°C. The newly hatched larvae measured 2.17 ± 0.29 mm in total length with a yolk volume of 0.165 ± 0.03 mm³ started feeding 36 hr after hatching. The present study, on induced breeding and chronological development of M. gulio embryo, will have significant implications on conservation and seed production for aquaculture.     

Effects of dietary protein on growth performance,nutrient utilization,digestive enzymes and physiological status of grey mullet,Mugil cephalus L. fingerlings reared in inland saline water

A 60 days feeding trial was conducted to illustrate the effect of graded levels of protein on the growth and metabolic enzymes of grey mullet (Mugil cephalus L.) fingerlings reared in inland saline water (ISW). Six isoenergetic (16 MJ/kg) and isolipidic (60 g/kg) diets containing 240, 260, 280, 300, 320 and 340 g crude protein (CP)/kg diet were formulated and fed to triplicate. Weight gain %, specific growth rate, protein utilizing efficiency, feed efficiency and RNA:DNA ratio were significantly higher (p < .05) in 320 and 300 g CP/kg diets. Fish fed with 240 g CP/kg diet showed significantly higher (p < .05) feed intake, whole‐body lipid content, hepatosomatic index value and liver glycogen content. Transaminase enzymes and malate dehydrogenase activities were elevated in fish fed 340 g CP/kg diet. Protease activity increased with increasing dietary CP level, but amylase activities showed an inverse relationship. No significant (p > .05) variations were observed for lactate dehydrogenase, oxidative stress enzymes, blood parameters and serum osmolality among all the treatment groups, but red blood cell count increases with increasing dietary CP levels. Based on the results, feeding dietary protein level of 300 g CP/kg is economically viable for rearing of grey mullet in ISW.     

Identification and pathogenicity study of emerging fish pathogens Acinetobacter junii and Acinetobacter pittii recovered from a disease outbreak in Labeo catla (Hamilton, 1822) and Hypophthalmichthys molitrix (Valenciennes, 1844) of freshwater wetland in West Bengal,India

The disease outbreaks in aquaculture system of wetlands are the major cause of fish mortality. Among various bacterial septicaemic diseases, fish mortality caused by Acinetobacter spp. is recently reported in different fish species. Fish disease outbreak was investigated in a wetland of West Bengal, India to identify the aetiological factors involved. The moribund fish were examined and subjected to bacterial isolation. Two bacterial causative agents were identified as Acinetobacter junii and Acinetobacter pittii by biochemical characterization and 16S rRNA gene amplification. Both the isolates were oxidase‐negative, nitrate‐negative, catalase‐positive and indole‐negative. The molecular identification using 16S rRNA gene sequencing and phylogenetic tree analysis further confirmed the two Acinetobacter spp. with 97%–99% similarity. The antibiotic resistance patterns of these two bacteria revealed that both of them were resistant to β‐lactam, cefalexin, cephalothin, amoxyclav, cefuroxime, cefadroxil, clindamycin, vancomycin and penicillin. In addition, A. pittii was also resistant to other antibiotics of cephams group such as ceftazidime and cefotaxime. In the challenge experiment, both A. junii and A. pittii were found to be pathogenic with LD₅₀ of 1.24 × 10⁵ and 1.88 × 10⁷ cfu/fish respectively. Histopathological examination of gill, liver and kidney revealed prominent changes supporting bacterial septicaemia. The investigation reports for the first time on concurrent infection by A. junii and multidrug‐resistant (MDR)‐A. pittii as emerging fish pathogens to cause severe mortality in Labeo catla and Hypophthalmichthys molitrix in a freshwater wetland.     

Fishmeal alternative from renewable CO2 for rainbow trout feed

The rapid global growth in fish farming and limited supply of fish meal (FM) has consequently reduced FM inclusion levels in compound feeds leading to a higher reliance on alternative protein sources. Sasya is a single cell protein (SCP) product that has a similar amino acid profile as FM. The aim of this study was to evaluate the effects of replacing FM with SCP on in vivo digestibility, growth, feed efficiency, whole‐body proximate/amino acid composition and gene expression levels of various hepatic enzymes in rainbow trout (Oncorhynchus mykiss). Three isonitrogenous (470 g/kg crude protein) and isolipidic (18 g/kg crude lipid) diets were formulated as follows: Diet 1: control (30% FM); Diet 2:24% FM + 6% SCP and Diet 3:18% FM + 12% SCP. Each diet was hand‐fed to triplicate tanks containing 30 rainbow trout fingerlings (4.99 ± 0.20 g) for 9 weeks. Apparent digestibility coefficients of SCP for dry matter, crude protein, lipid and energy were 60, 80, 93 and 74% respectively. Growth performance (final weight: 69–71 g), feed conversion ratios (0.91–0.94) as well as whole‐body protein and amino acid composition were unaffected by diets. However, Diet 3 significantly increased whole‐body crude fat and energy. Fish fed the SCP‐based diets had significantly higher expression for carnitine palmitoyltransferase‐1b (CPT1b), fatty acid delta 6 desaturase (FADS6) and fatty acid elongase 5 compared to the control. Overall, the quality of the SCP was similar as FM. Therefore, this product could enlarge the portfolio of alternative protein sources that can be used in fish diets and thus open a new market opportunity for use of a new feed resource in the feed industry.