On a new species of Pseudocapillaria (Nematoda: Trichuroidea), P. lepidocephali,from the estuarine fish Lepidocephalus (Lepidocephalichthys) guntea (Hamilton)

Pseudocapillaria lepidocephali n. sp., parasitic in the intestine of the loach Lepidocephalus (Lepidocephalichthys) guntea from the Hooghly estuary at Kalyani, West Bengal, India, is described. The parasite is characterised mainly by its small body size, the structure of the male caudal end (the presence of mediumsized ventro-lateral lobes and the absence of a dorsal membrane), the presence of a non-spinous spicular sheath, the structure of the stichosome (26–34 stichocytes present), and the size and structure of the eggs. This is the first species of the genus Pseudocapillaria described from India and also from the genus Lepidocephalus.     

A bioenergetics model for an air-breathing fish, Channa striatus

Food consumption, standard metabolism, and growth of juvenile snakehead, Channa striatus, a cannibalistic and air-breathing fish were measured at 24–26 °C under controlled laboratory condition. Snakehead weighing 3.2–29.5 g were evaluated, and were fed smaller snakehead. Based on our observations, we determined bioenergetics relationships between specific food consumption, metabolic rates, and body weight. These values, along with other published parameter values allowed us to construct a bioenergetics model for snakehead. We then verified our model with growth and food consumption measurements from an independent feeding trial. Predicted fish growth closely matched observed growth. Our model underestimated cumulative food consumption when a constant activity value was used, but consumption estimates improved when we used non-constant activity values (1-5 times of standard metabolism). Predicted fish maintenance ration was 1.7% of body weight per day. Food conversion efficiency was greatest (0.59) when fed 2% body weight daily, but declined when daily consumption exceeded 6% body weight. This model provides a useful approach for assessing food requirements of snakehead under controlled condition. This revised version was published online in July 2006 with corrections to the Cover Date.     

Seasonal variations in the blood constituents of an air-breathing fish, Channa punctatus Bloch

A year long study based on monthly observations on the haematology of female Channa punctatus with respect to haemoglobin, haematocrit values, total erythrocyte and leucocyte numbers together with a differential enumeration of various leucocytes viz. thrombocytes small and large lymphocytes, monocytes, neutrophils, eosinophils and basophils is recorded. The study showed that the total erythrocytic number, haemoglobin and haematocrit values decrease during the breeding time i.e. July to September. The thrombocyte number significantly increased while the neutrophilic number showed a decrease. The physiological significance of these changes is discussed with reference to the available literature.     

Retinopetal neuronal system in the brain of an air-breathing teleost fish,Channa punctata

Summary Retinopetal neurons were visualised in the telencephalon and diencephalon of an air-breathing teleost fish, Channa punctata, following administration of cobaltous lysine to the optic nerve. The labelled perikarya (n=45–50) were always located on the side contralateral to the optic nerve that had received the neuronal tracer. The rostral-most back-filled cell bodies were located in the nucleus olfactoretinalis at the junction between the olfactory bulb and the telencephalon. In the area ventralis telencephali, two groups of telencephaloretinopetal neurons were identified near the ventral margin of the telencephalon. The rostral hypothalamus exhibited retrogradely labelled cells in three discrete areas of the lateral preoptic area, which was bordered medially by the nucleus praeopticus periventricularis and nucleus praeopticus, and laterally by the lateral forebrain bundle. In addition to a dorsal and a ventral group, a third population of neurons was located ventral to the lateral forebrain bundle adjacent to the optic tract. The dorsal group of neurons exhibited extensive collaterals; a few extended laterally towards the lateral forebrain bundle, whereas others ran into the dorsocentral area of the area dorsalis telencephali. A few processes extended via the anterior commissure into the telencephalon ipsilateral to the optic nerve that had been exposed to cobaltous lysine. However, the ventral cell group did not possess collaterals. In the diencephalon, retinopetal cells were visualised in the nucleus opticus dorsolateralis located in the pretectal area; these were the largest retinopetal perikarya of the brain. The caudal-most nucleus that possessed labelled somata was the retinothalamic nucleus; it contained the largest number of retinopetal cells. The limited number of widely distributed neurons in the forebrain, some with extensive collaterals, might participate in functional integration of different brain areas involved in feeding, which in this species is influenced largely by taste, not solely by vision.     

Responses of aerial ventilation to hypoxia and hypercapnia inChanna argus,an air-breathing fish

Summary The snake-head fish (Channa argus) is an obligate air-breather inhabiting fresh waters in the temperate zone of East Asia.Ventilation of the air-breathing organ and aerial gas exchange were measured in 1 to 2 kg specimens at 15 and 25°C. Additionally, the ventilatory responses to hypoxia and hypercapnia were studied. Aerial ventilation increased from 1.1 to 2.9 mlbtps·kg^–1·min^–1 when temperature rose from 15 to 25°C. Concomitantly, O₂-uptake through airbreathing increased from 0.1 mlstpd·kg^–1·min^–1 (15°C) to 0.28 mlstpd·kg^–1·min^–1 (25°C), whereas aerial gas exchange was less important for CO₂-climination as evident from low gas exchange ratios (0.16 at 15°C, 0.29 at 25°C).Ventilation increases only slightly in response to inspiration of hypercapnic gas mixtures or to hypoxic conditions in water. By contrast, inspiration of hypoxic gas mixtures caused marked increases of ventilation in particular at the higher temperature.Aerial ventilation inChanna is low compared to values for ectothermic pulmonary breathers. However, its ventilatory responses to hypoxia strikingly resemble those of reptiles: The most marked ventilatory response to hypoxia occurs at the higher temperature where the demands for O₂ are greatest.     

Cell types in the peripheral blood of an air-breathing fish Channa punctatus

Different cell types found in the peripheral blood of Channa punctatus , an air-breathing freshwater fish have been characterized and identified using morphological, morphometric, cytochemical and autoradiographic techniques. Some of the cytochemical methods used, particularly that of differential staining of haemoglobin have been developed and used for the first time in fishes.     

Toxicity of mercury: A comparative study in air-breathing and non air-breathing fish

Annual primary production was estimated from two years data. The results indicate a high primary productivity and a short turn over time for nitrogen.

     

Haematological and haematopoietic responses to starvation in an air-breathing fish Channapunctatus Bloch

Changes in haematological values (RBC numbers, haemoglobin content, haematocrit value, MCV, MCH, MCHC, TLC and DLC) based on weekly samples from a group of starved fish were investigated. After 8 weeks of starvation, the effects of restoration to a normal diet was evaluated. Parallel studies on haematopoietic tissues were also made. Changes in some biochemical values such as blood glucose, liver and muscle glycogen were also examined to correlate biochemical effects with those of haematological changes. Erythrocytes, thrombocytes and neutrophils were found to be most sensitive to starvation. The initial response to deprivation of food was an increase in RBCs and related values and in total leukocyte population. However, from week 5 onwards a sharp decline in these cell populations was noted. The leukocytes and thrombocytes showed a change parallel to RBC and the total leukocyte counts. However, neutrophils were observed to show a consistent increase throughout the starvation period. A blood glucose level below SOmglOOmh¹ appeared critical in relation to blood cell population. Haematopoietic studies revealed that reticulocytes and mesomyelocytes were unable to keep pace with the changing peripheral blood picture. Other stages in development responded to the changes in the peripheral blood.     

Haematological and haematopoietic response to acid stress in an air-breathing freshwater fish, Channa punctatus Bloch

Response of Channa punctatus to acidic water was studied by exposing fishes to pH 3.5, 4.5, 5.5 and 6.5 for 6 weeks. Growth and mortality data indicated increasing stress as the acid level in the ambient water increased. While no mortality was recorded at pH 6.5, a distinct loss of weight compared to continuous gain in body weight in control fish indicated stress. As the pH level decreased, the rate of loss in body weight increased accompanied by mortality which rose to as high as 60% within 3 weeks in fishes exposed to water at pH 3.5.
Haematological investigations confirmed the general stress indicated by growth and mortality data. Thus. RBC and related values indicated overall polycythemia. However, eosinophils, basophils. and large and small lymphocytes showed a distinct fall in number as compared to the control.
Correlated haematopoietic studies revealed that both the initial and penultimate stages in RBC and neutrophil development recorded an increase parallel to that observed in peripheral blood, but intermediate stages, probably because they were unable to keep pace with the fast turnover, showed a relative decrease.
Biochemical investigations showed an increase not only in blood glucose level but also in liver glycogen content. However, there was a significant decrease in muscle glycogen reserves.
The significance of these changes is discussed.     

Haematological, haematopoietic and biochemical responses to thermal stress in an air-breathing freshwater fish, Channa punctatus Bloch

Results of experiments on fishes maintained at three different temperatures (25, 30 and 35° C), compared with a control group maintained at 14° C, are presented. Fishes showed obvious signs of stress at 30 and 35° C, as indicated by loss of weight and increase in mortality rate. On the other hand, those at 14 and 25° C gained weight steadily and showed normal growth.
Biochemical studies confirm the stress symptoms, as there was a steady, statistically significant fall in blood glucose level and depletion of glycogen reserves in liver and muscle within the first week at 35° C and by the fourth week at 30° C.
These results were further confirmed by haematological investigations. All the parameters tested indicated deviation from the normal healthy conditions seen at 14 and 25° C. The deviations in most of the parameters at 35° C were statistically significant from about the second week, and at 30° C by the fourth week. Erythrocytic polycythemia accompanied by an increase in haemoglobin content and haematocrit values were indicative of thermal stress. Leucopenia was mainly contributed by neutrophil decrease, but a relative increase in thrombocytes was recorded.
Correlated erythropoietic studies indicated an increase both in the initial (small lymphoid haemoblast) and penultimate (mature reticulocyte) stages in red cell development, although the intermediate stages decreased in number. Similar trends in the different stages of neutrophil development were observed. The significance of these changes in the context of homeostatic phenomena is discussed.     

Changes in the blood parameters of an air-breathing fish during different respiratory conditions.

Some of the blood parameters recorded in an air-breathing eel, Amphipnous cuchia under normal respiratory condition during non-breeding period (September-April) are haemoglobin (Hb) concentration 19.26%, haematocrit value 56.16%, RBC number 1.71 million/mm3, RBC size 18.86 X 9.70 mum, mean corpuscular haemoglobin (MCH) 113.4 ng, mean corpuscular haemoglobin concentration (MCHC) 34.2%, blood sugar 77 mg% and ascorbic acid 0.435 mg%. The higher concentration of haemoglobin (19.26%) appears to be related to its obligatory air breathing habit and habitat in a water of low oxygen content. Though a definite trend of increase in the haemoglobin and haematocrit concentration with an increase in the body weight of the fish was lacking, variations were clearly marked related to intrinsic activity of the fish connected with different respiratory conditions. Asphyxiation in a submerged but continuous flow of water (liter/h) for 5 1/2 h resulted in an increase in the above-mentioned parameters to an appreciable extent. These increases were 0.23 million/mm3 in the number of erythrocytes, 6.16% in haemoglobin concentration, 10% in haematocrit value, 20% in blood sugar and 35% in ascorbic acid content. The mean corpuscular haemoglobin showed a decline of 6.2%. Exclusive aerial breathing for 5 1/2 h also caused 7.4% increase in haemoglobin concentration, 9.4% in haematocrit value, 0.14 million/mm3 in RBC number, 20% in blood sugar level, 9% in ascorbic acid content but almost no change in mean corpuscular haemoglobin. The average surface area for diffusion of gases appeared to have reduced by 6.8 mum2 per RBC.     

The source of lamellar mitochondria-rich cells in the air-breathing fish, Trichogaster leeri

Pavement cells and the mitochondria-rich cells (MRCs) are two of the main types of cells in fish gill epithelia. The pavement cells are generally responsible for gas exchange and MRCs for ion regulation. MRCs are found especially in the trailing edge and the interlamellar region of gill filament. In some species, MRCs are also observed in the gill lamellae. A previous study reported the likelihood of having lamellar MRCs in air-breathing fishes. Nevertheless, the source of lamellar MRCs is unclear. We used the air-breathing fish, Trichogaster leeri, to investigate the source of proliferated cells on the lamellae when 5-bromo-2-deoxyuridine (BrdU) was injected at different times before fish were sampled from deionized water. There were two major findings in this study. First, undifferentiated cells were found in the lamellae, as well as in the filaments. And, within 12-24 hr, a proliferated cell, identified as BrdU cell, could differentiate to an MRC in the gill lamellae. Second, the filaments and the lamellae in T. leeri responded to ionic stress differently but the proportion of the proliferated MRCs to the BrdU cells remained constant. Our results suggested that the lamellar MRCs were mainly differentiated from the cells that proliferated earlier from the lamellae.     

Sublethal malathion induced changes in the ovary of an air-breathing fish,Heteropneustes fossilis: a histological study

This paper describes effects of a sublethal (1.2 mg 1^–1) organophosphate, malathion, on the ovary of an air breathing catfish, Heteropneustes fossilis. The study focuses on microscopic changes that occur on ovigerous lamellae, oocytes at different stages of development and the nucleus of the immature oocyte. Also, change in estrogen levels in blood serum is investigated. Clumping of cytoplasm appears after 24 h of exposure to malathion. Clumping intensified after 48 h. Degeneration in the follicular cells was also observed. After 72 h exposure the number of nucleoli increased, nuclear materials shrunk, oocytes became adhered. With 96 h of exposure, nuclear materials of all the oocytes shrunk to a smaller clump. The oocytes fused together, and follicular epithelium became loose and ruptured. A few atretic oocytes were visible. Radioimmunoassay of the estrogen level in blood serum after 72 h of exposure of malathion showed a reduction in the level. This study showed that the histopathological condition of the gonad is reflected in malfunctioning of the endocrine system and hormonal disbalance.     

Effects of feeding and starvation on growth and swimming activity in an obligatory air-breathing fish

Reared in (tubular) aquaria containing different depths of water, Ophiocephalus striatus (0.7 g, 4.5 cm body length), an obligatory air-breathing tropical fish, swam long or short distances to enable themselves to exchange atmospheric air. In each tested depth (2.5, 5.0, 15.5, 31.0 and 40.0 cm) series, one group was starved, while the other was fed ad libitum twice a day on fish muscle. In the shallowest water (2.5 cm depth), the feeding group surfaced 1,294 times, travelling 64.7 m at an energy cost of 20.4 mg dry fish substance/g live fish/day, against those exposed to the deepest water (40 cm depth), which expended 35.8 mg/g/day, swimming 1,503.4 m on 1,879 visits to the surface. The starving group surfaced only 482 times, travelling 24.1 m at an expense of 5.8 mg/g/day in the shallowest water, while those at 40 cm depth surfaced 504 times, swimming 403.2 m at an energy cost of 7.4 mg/g/day. Owing to the sustained swimming activity and the consequent fatigue, the test individuals belonging to both groups in all the tested series hang to the surface for a definite interval, repaying the O₂ debt. Observations were also made to assess the duration of hanging to precisely estimate the distance travelled. Irrespective of changes in depths of water, the duration of hanging to surface was only 3.0 hr/day for the feeding groups, while it was as much as 15.5 hr/day for the starving groups. The maximum sustained metabolic level of O.striatus reared in 40 cm depth was equivalent to 1.23 ml O₂/g/hr, which is about 2 times higher than the value reported for the active metabolism of swimming Oncorhynchus nerka at 15°C in Brett's (1964) respirometer. O.striatus reared in 2.5 cm depth fed 32.0 mg and converted 6.7 mg dry food/g live fish/day, while those exposed to the deepest water fed 49.1 mg, but converted only 5.5 mg/g/day. Culturing obligatory air-breathing fishes in shallow waters will be advantageous.     

Vasculature of the head and respiratory organs in an obligate air-breathing fish,the swamp eel Monopterus (=Amphipnous) cuchia

Methyl methacrylate vascular corrosion replicas were used to examine the macrocirculation in the head region and the microcirculation of respiratory vessels in the air-breathing swamp eel Monopterus cuchia. Fixed respiratory tissue was also examined by SEM to verify capillary orientation. The respiratory and systemic circulations are only partially separated, presumably resulting in supply of mixed oxygenated and venous blood to the tissues. A long ventral aorta gives rise directly to the coronary and hypobranchial arteries. Two large shunt vessels connect the ventral aorta to the dorsal aorta, whereas the remaining ventral aortic flow goes to the respiratory islets and gills. Only two pairs of vestigial gill arches remain, equivalent to the second and third arches, yet five pairs of aortic arches were identified. Most aortic arches supply the respiratory islets. Respiratory islet capillaries are tightly coiled spirals with only a fraction of their total length in contact with the respiratory epithelium. Valve-like endothelial cells delimit the capillary spirals and are unlike endothelial cells in other vertebrates. The gills are highly modified in that the lamellae are reduced to a single-channel capillary with a characteristic three-dimensional zig-zag pathway. There are no arterio-arterial lamellar shunts, although the afferent branchial artery supplying the gill arches also supplies respiratory islets distally. A modified interlamellar filamental vasculature is present in gill tissue but absent or greatly reduced in the respiratory islets. The macro- and micro-circulatory systems of M. cuchia have been considerably modified presumably to accommodate aerial respiration. Some of these modifications involve retention of primitive vessel types, whereas others, especially in the microcirculation, incorporate new architectural designs some of whose functions are not readily apparent.     

Physico-chemical characterisation and molecular organisation of the collagen from the skin of an air-breathing fish (Ophiocephalus striatus)

Collagen has been prepared from the skin of an air-breathing Indian fish(Ophiocephalus striatus) by extraction with cold 0.5M acetic acid and purification by alternate precipitation with NaCl and dialysis against 0.02M Na₂HPO₄. The purified collagen was characterised with respect to physico-chemical properties, amino acid composition and chromatography of the denatured collagen. The fish collagen has a higher shrinkage temperature and denaturation temperature compared to that of the allied teleosts living in exclusively aquatic medium. These differences could possibly be reflections of the response to the rigours of the environment. As found for other vertebrate collagens, the fish collagen contains two kinds of single chains the α₁ and α₂ chains as revealed by sodium dodecyl sulphate-polyacrylamidegel electrophoresis and carboxymethylcellulose chromatography.     

Nucleoprotein cytochemistry during oogenesis in a unisexual fish, Poecilia formosa

Summary Cytochemical methods and electron microscopy were used to study changes in the chemical composition of nuclear, nucleolar and perinuclear bodies during the early stages of oocyte development inPoecilia formosa, an apomictic species of fish that produces only female offspring. Prominent accumulations of ribonucleoprotein (RNP) occur in nucleoli and appear on either side of the nuclear envelope during diplotene. In certain planes of section, RNP material seems to be in transit across this interface.En bloc acid extractions or RNAse treatment abolished basophilia and markedly reduced the electron density of both nucleoli and cytoplasmic nucleolar-like bodies. DNA-specific fluorescent probes such as mithramycin failed to reveal nucleolar cores in poeciliid oocytes, although the same procedures showed unequivocal localization of GC-rich DNA cores within multiple nucleoli of diplotene oocytes fromXenopus laevis or the rainbow trout,Salmo gairdneri. Also, cytological hybridization studies, utilizing [³H]rRNA as a probe for nucleolar oocytes. Feulgen-stained pachytene oocytes ofP. formosa have twice the number of chromosome strands seen in similar stages of oocytes from two, related bisexual species,P. mexicana andP. latipinna. Although the bivalent nature of these chromosomes could not be resolved with the light microscope, configurations resembling, but not identical to, synaptonemal complexes were identified by electron microscopy.     

The effect of acidity on gill variations in the aquatic air-breathing fish,Trichogaster lalius

Climate change affects organisms that inhabit not only in aerial but also in aquatic environments by making water more hypoxic and acidic. In the past, we evaluated morphological and functional variations in the gills of 12 species of aquatic air-breathing fishes. The aim of the present study is to examine the degree of gill modification in the aquatic air-breathing fish, Trichogaster lalius, in response to acidic stress. This provides a link between the ecological and physiological studies. We evaluated the changes in morphology and function of the gills, labyrinth organ, and kidney when the fish were subjected to acidic water and deionized water (DW). In the first experiment, fish were sampled at 1, 2, 4, and 7 days after acidic treatment. Apparent morphological modification was observed on day 4 and recovery was noted on day 7. Protein expression and enzyme activity of vacuolar-type H⁺-ATPase (VHA) and the protein expression of the proliferating cell nuclear antigen (PCNA) of the 1st and 4th gill arches both increased in the 4-day and 7-day acidic groups while the enzyme activity of Na⁺/K⁺-ATPase (NKA) decreased. In the second experiment, fish were tested for changes in the 1st and 4th gill arches and kidney after exposure to DW and acidic water for 4 days. The gill structure of the fish in the DW was not different from that of the control group (fresh water). The protein expression and enzyme activity of the VHA of the 1st and 4th gill arches increased in both the DW and acidic groups for 4 days. We found a decrease in the protein expression of NKA in the kidney and in the enzyme activity of NKA in the 1st and 4th gill arches in the DW and acidic groups. From these results, we suggest that T. lalius exhibited significantly different ionic regulation and acid-base regulatory abilities in the DW and acidic groups in the 1st and 4th gill arches and kidney. The responses of the gills in T. lalius were different from those fish that show apparent morphological variations between the 1st and 4th gill arches.