Ammonia is an unusual toxicant in that it is produced by, as well as being poisonous to, animals. In aqueous solution ammonia has two species, NH3 and NH4+, total ammonia is the sum of [NH3] + [NH4+] and the pK of this ammonia/ammonium ion reaction is around 9.5. The NH3/NH4+ equilibrium both internally in animals and in ambient water depends on temperature, pressure, ionic strength, and pH; pH is most often of greatest significance to animals. Elevated ammonia levels in the environment are toxic. Temperature has only minor effects on ammonia toxicity expressed as total ammonia in water, and ionic strength of the water can influence ammonia toxicity, but pH has a very marked effect on toxicity. Acid waters ameliorate, whereas alkaline waters exacerbate ammonia toxicity. The threshold concentration of total ammonia ([NH3] + [NH4+]) resulting in unacceptable biological effects in freshwater, promulgated by the EPA (1998), is 3.48 mg N/liter at pH 6.5 and 0.25 mg N/liter at pH 9.0. There is only a relatively small saltwater data set, and a paucity of data on ammonia toxicity in marine environments, particularly chronic toxicity. The national criteria promulgated in the EPA (1989) saltwater document is a criterion continuous concentration (chronic value) of 0.99 mg N/liter total ammonia and a criterion maximum concentration (half the mean acute value) of 6.58 mg N/liter total ammonia, somewhat less than the equivalent freshwater pH 8.0 values of 1.27 and 8.4 mg N/liter total ammonia, respectively. This is consistent with marine species being somewhat more sensitive to ammonia than freshwater species. Toxicity studies are usually carried out on unfed, resting fish in order to facilitate comparison of results. Based on recent studies, however, environmental stresses, including swimming, can have dramatic effects on ammonia toxicity. It is also clear that feeding results in elevated postprandial body ammonia levels. Thus, feeding will probably also exacerbate ammonia toxicity. Fish may be more susceptible to elevated ammonia levels during and following feeding or when swimming. Thus, present ammonia criteria may fail to protect migrating fish and may be inappropriate for fish fed on a regular basis. Most teleost fish are ammonotelic, producing and excreting ammonia by diffusion of NH3 across the gills. They are very susceptible to elevated tissue ammonia levels under adverse conditions. Some fish avoid ammonia toxicity by utilizing several physiologic mechanisms. Suppression of proteolysis and/or amino acid catabolism may be a general mechanism adopted by some fishes during aerial exposure or ammonia loading. Others, like the mudskipper, can undergo partial amino acid catabolism and use amino acids as an energy source, leading to the accumulation of alanine, while active on land. Some fish convert excess ammonia to less toxic compounds including glutamine and other amino acids for storage. A few species have active ornithine—urea cycles and convert ammonia to urea for both storage and excretion. Under conditions of elevated ambient ammonia, the mudskipper P. schlosseri can continue to excrete ammonia by active transport of ammonium ions. There are indications that some fish may be able to manipulate the pH of the body surface to facilitate NH3 volatilization during aerial exposure, or that of the external medium to lower the toxicity of ammonia during ammonia loading. Future investigation of these aspects of “environmental ammonia detoxification” may produce new information on how fish avoid ammonia intoxication.

Ammonia toxicity, tolerance, and excretion

Many fishes are ammonotelic but some species can detoxify ammonia to glutamine or urea. Certain fish species can accumulate high levels of ammonia in the brain or defense against ammonia toxicity by enhancing the effectiveness of ammonia excretion through active NH4+ transport, manipulation of ambient pH, or reduction in ammonia permeability through the branchial and cutaneous epithelia. Recent reports on ammonia toxicity in mammalian brain reveal the importance of permeation of ammonia through the blood-brain barrier and passages of ammonia and water through transporters in the plasmalemma of brain cells. Additionally, brain ammonia toxicity could be related to the passage of glutamine through the mitochondrial membranes into the mitochondrial matrix. On the other hand, recent reports on ammonia excretion in fish confirm the involvement of Rhesus glycoproteins in the branchial and cutaneous epithelia. Therefore, this review focuses on both the earlier literature and the up-to-date information on the problems and mechanisms concerning the permeation of ammonia, as NH3, NH4+ or proton-neutral nitrogenous compounds, across mitochondrial membranes, the blood-brain barrier, the plasmalemma of neurons, and the branchial and cutaneous epithelia of fish. It also addresses how certain fishes with high ammonia tolerance defend against ammonia toxicity through the regulation of the permeation of ammonia and related nitrogenous compounds through various types of membranes. It is hoped that this review would revive the interests in investigations on the passage of ammonia through the mitochondrial membranes and the blood-brain barrier of ammonotelic fishes and fishes with high brain ammonia-tolerance, respectively.

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Ammonia production, excretion, toxicity, and defense in fish: a review.

The branchial epithelium of the mudskipper Periophthalmodon schlosseri is densely packed with mitochondria-rich (MR) cells. This species of mudskipper is also able to eliminate ammonia against large inward gradients and to tolerate extremely high environmental ammonia concentrations. To test whether these branchial MR cells are the sites of active ammonia elimination, we used an immunological approach to localize ion-transport proteins that have been shown pharmacologically to be involved in the elimination of NH(4)(+) (Na(+)/NH(4)(+) exchanger and Na(+)/NH(4)(+)-ATPase). We also investigated the role of carbonic anhydrase and boundary-layer pH effects in ammonia elimination by using the carbonic anhydrase inhibitor acetazolamide and by buffering the bath water with Hepes, respectively. In the branchial epithelium, Na(+)/H(+) exchangers (both NHE2- and NHE3-like isoforms), a cystic fibrosis transmembrane regulator (CFTR)-like anion channel, a vacuolar-type H(+)-ATPase (V-ATPase) and carbonic anhydrase immunoreactivity are associated with the apical crypt region of MR cells. Associated with the MR cell basolateral membrane and tubular system are the Na(+)/K(+)-ATPase and a Na(+)/K(+)/2Cl(-) cotransporter. A proportion of the ammonia eliminated by P. schlosseri involves carbonic anhydrase activity and is not dependent on boundary-layer pH effects. The apical CFTR-like anion channel may be serving as a HCO(3)(-) channel accounting for the acid-base neutral effects observed with net ammonia efflux inhibition.

Immunolocalization of ion-transport proteins to branchial epithelium mitochondria-rich cells in the mudskipper (Periophthalmodon schlosseri)

There are a small number of fish species, both marine and freshwater, that exhibit a truly amphibious habit that includes periods of aerial exposure. The duration of emersion is reflected in the level of physical and physiological adaptation to an amphibious lifestyle. Fish that are only briefly out of water retain predominantly aquatic attributes whereas there are semi-terrestrial species that are highly adapted to prolonged periods in the aerial habitat. Desiccation is the main stressor for amphibious fish and it cannot be prevented by physiological means. Instead, amphibious fish resist excessive water loss by means of cutaneous modification and behavioural response. The more terrestrially adapted fish species can tolerate considerable water loss and may employ evaporation to aid thermoregulation. The amphibious habit is limited to fish species that can respire aerially. Aerial respiration is usually achieved through modification to existing aquatic pathways. Freshwater air-breathers may respire via the skin or gills but some also have specialized branchial diverticula. Marine species utilize a range of adaptations that may include modified gills, specialized buccopharyngeal epithelia, the intestine and the skin. Areas of enhanced respiratory activity are typified by increased vascularization that permits enhanced perfusion during aerial exposure. As with other adaptations the mode of nitrogenous elimination is related to the typical durations of emersion experienced by the fish. Intertidal species exposed on a regular cycle, and which may retain some contact with water, tend to remain ammoniotelic while reducing excretion rates in order to prevent excessive water loss. Amphibious fish that inhabit environments where emersion is less predictable than the intertidal, can store nitrogen during the state of emersion with some conversion to ureotelism or have been shown to tolerate high ammonia levels in the blood. Finally, the more amphibious fish are more adapted to moving on land and seeing in air. Structural modifications to the pectoral, pelvic, dorsal and anal fins, combined with a well-developed musculature permit effective support and movement on land. For vision in air, there is a general trend for fish to possess close-set, moveable, protruberant eyes set high on the head with various physical adaptations to the structure of the eye to allow for accurate vision in both air and water.

Adaptations of amphibious fish for surviving life out of water

Periophthalmodon schlosseri can maintain ammonia excretion rates and low levels of ammonia in its tissues when exposed to 8 and 30 mM NH4Cl, but tissue ammonia levels rise when the fish is exposed ...

The mudskipper, Periophthalmodon schlosseri, actively transports NH 4 + against a concentration gradient

The three mudskippers—Periophthalmus chrysospilos, Boleophthalmus boddaerti, and Periophthalmodon schlosseri—occupy the same macrohabitat in Singapore but have different behaviors. Correlations were made between differences in behavior and morphological adaptations of their gills to tolerate terrestrial exposure.P. schlosseri has branched gill filaments, thick gill rods, and fused secondary lamellae which enable them to better adapt to a terrestrial than an aquatic environment. Of the three mudskippers, P. chrysospilos gills are the shortest. They are also bent and poorly developed for aquatic respiration. B. boddaerti gills consist of numerous long filaments and have the largest gill area of the three mudskippers. These features suggest that B. boddaerti gills function more efficiently as a respiratory organ in water than in air.Gill surfaces of all three mudskippers are highly convoluted to increase surface area.

A comparative study of terrestrial adaptations of the gills in three mudskippers-Periophthalmus chrysospilos, Boleophthalmus boddaerti, and Periophthalmodon schlosseri

D. J. W. Lane

Papers

A comparative study of terrestrial adaptations of the gills in three mudskippers-Periophthalmus chrysospilos, Boleophthalmus boddaerti, and Periophthalmodon schlosseri