THE FUNCTIONAL ANATOMY OF THE DIGESTIVE TRACT OF A SHRIMP METAPENAEUS BENNETTAE RACEK & DALL* (CRUSTACEA : DECAPODA : PENAEIDAE)

[Manuscript received October 17, 19661 Summary

The anatomy of the proventriculus, digestive gland, midgut and its diverticula, and the rectum is described. In structure and function the proventriculus is similar to that of anumber of other Decapoda. Two distinct cell types occur in the digestive gland, a secretory type, and a mucopolysaccharide-containing type, whose function is not clear. The digestive gland has no intrinsic muscles, and depends on extrinsic muscles, and possibly ingested water, for filling and emptying. The midgut extends to the sixth abdominal somite and faecal material is contained in a peritrophic membrane. Evidence for secretory functions of the midgut and anterior and posterior diverticula is discussed. The rectal lining is formed into six longitudinal pads which are used to expel long sections of peritrophic membrane containing faeces. Methods of feeding are described. Permeability of the anterior proventriculus to 22Na and [14C] glucose was measured; 22Na approached equilibrium in.6-7 hr, but [14C]glucose passed through at about one-seventh this rate, indicating that direct glucose uptake from the proventriculus would be negligible. Food, labelled with particulate llOAg,was found to begin leaving the proventriculus almost as soon as it was filled, but complete emptying took 6-12 hr. Defecation was at a peak 5-8 hr after food ingestion, but continued up to 20 hr. The rectum appears to have the additional function of pumping water into the gut via the anus.

While detailed studies of the functional anatomy of the digestive tract of a number of Decapoda Reptantia have been carried out, e.g. Astacus (Huxley 1884; Balss 1927), Cancer (Pearson 1908), Nephrops norvegicus (Yonge 1924), Galathea (Pike 1947), knowledge of Natantia is meagre. Patwardan ( 1 9 3 5 ~ ~ 1935b) described the structure and mechanism of the gastric mills of various shrimp, and Pillai (1960) studied the whole digestive system of Caridina laevis. Although there have been some physiological investigations of Penaeidae (MacFarland and Lee 1963; Dall 1964, 1965a-1965d), none has dealt specifically with the digestive tract. Dall(1965c) treated some aspects of carbohydrate content of the digestive gland and also (1965d) presented evidence for excretion of calcium via the anus. Other studies have been confined to purely morphological features. Kubo (1949) and Dall (1957) used the "stomodoeal apparatus" for taxonomic features, and although Young (1959) published a monograph on the morphology of Penaeus setiferus, most of it concerns musculature and the section on the gut is brief.

* Formerly M. mastersii (Haswell) (see Racek and Dall 1965). t Department of Zoology, University of Queensland, Brisbane;
of Guelph, Ontario, Canada.

present address: University

Aust. J. Zool., 1967, 15, 699-714

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In the present investigation attention has been confined to morphology and histology of the gut, and the manner in which the food is treated by the respective regions. No study of digestion and absorption, apart from proventriculus permeability, has been made. Nevertheless, from the diverse nature of food eaten, it would seem that, as in Nephrops, a full complement of the usual enzymes is probably present (Yonge 1924). The musculature and armature of the proventriculus in Penaeidae have been described in detail by Kubo (1949) and Young (1959), and only the histology and overall functions are dealt with here. The mouthparts of Metapenaeus are similar to those of most other genera within the family, and are not described here (see Kubo 1949 (various genera); Young 1959 (Penaeus setiferus)).

Animals were collected by trawling in Moreton Bay, Queensland. All were adults of 14-20-mm carapace length (Dall 1958), and only actively feeding intermoult shrimp were used. Holding tanks were maintained at 20-25OC, with salinity 35%,. Food was normally given two or three times per week, but no food was given for 2-4 days prior to experiments. All experiments were carried out at 20C, with water salinity 35%,, unless otherwise stated. Tissues were always fixed directly from living animals. General fixatives used were formol-saline, Heidenhain's Susa, Zenker (Pantin 1946). For cytological detail of digestive gland, midgut and its diverticula, an iso-osmotic, osmium tetroxide, buffered fixative was used consisting of 5 ml barbital buffer; 16 ml filtered sea water of salinity 35%,; 5 ml 0 . IN HCl; and 0.2 g 0 s 0 4 ; final pH of solution 7.4; fixation temperature 4C (modified from Pease 1960). Tissues were fixed for 20-30 min. Formol-saline at 5OC gave best results for the digestive gland, which is notoriously difficult to fix before autolysis begins, which may cause artifacts giving the appearance of sloughed-off cells. Good fixation was obtained with the osmium tetroxide fixative only with pieces of tissue less than 1 mm thick. All tissues were embedded in paraffin and cut at 7-10 ,u. Stains employed were Mayer's haemalum, Mallory's (Pantin 1946); histochemical methods used were Feulgen reaction for DNA, periodic acidSchiff (P.A.S.), and toluidine blue for polysaccharides (Pearse 1961). The Radiochemical Centre, Amersham, supplied Z2NaCl and [W]glucose as stock items, and llOAg was obtained from the Australian Atomic Energy Commission. Specific activity of the latter was about 1 mc per 400 mg AgN03. Finely divided silver was produced by reducing a dilute solution with hydroquinone and thoroughly washing the precipitate. Particle size ranged up to 10 p, but most particles were 1-5 p. Counting was carried out with an EKCO 645A ratemeter with scintillation head, or with an EKCO 664A scintillation counter and N530F autoscaler.

(a) Anatomy of the Digestive Tract Figure 1 shows the general organization of the gut. A short oesophagus opens vertically into a large anterior chamber, followed by a smaller posterior chamber

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partly surrounded by the digestive gland. The cuticular lining of the foregut ends at the junction of the posterior chamber and midgut. The tubular midgut extends from the thorax to the anterior region of the sixth abdominal somite, and like the anterior and posterior diverticula, is lined by columnar epithelium. Proctodoeal cuticle lines the rectum up to its junction with the midgut and posterior diverticulum.

Fig. 1.-Lateral dissection of the complete digestive tract. AC, Anterior chamber of proventriculus; anterior diverticula; pco, opening of posterior chamber of proventriculus into digestive gland; MG, midgut; PD,posterior diverticulum; R, rectum; A, anus; T, telson; MO, mouth; o, oesophagus; a-a, 6-b, c-c, d-d, e-e, represent the planes of the transverse sections shown in Figures 2, 4, 5, 8, and 7, respectively.
AD,

(i) Proventriculus (Figs. 2-5) The terminology of this structure is in an unsatisfactory state. Although Pearson (1908) objected to the use of "cardiac" and "pyloric" for anterior and posterior chambers respectively, this misleading nomenclature has persisted. The use of the term "stomach" for the anterior chamber is also to be condemned; the stomach functions more as a crop. Kubo (1949) referred to the complete foregut structure as the "stomodoeal apparatus", but the less clumsy "proventriculus" has been more generally accepted. As indicated in Figure 1, the chamber into which the oesophagus opens, and which has a function analogous to the crop of many insects, is referred to here as the "anterior chamber" (Fig. 2). The posterior opening of this chamber is closed by the complex armature of the gastric mill (Kubo 1949; Dall 1957). Triturated food passes into the dorsal compartment of the posterior chamber (Fig. 4). Only the finest particles are able to pass via the complex setose "presses" of the ventral compartment to the openings of the digestive gland (Fig. 5). Larger particles which are excluded are apparently free to pass directly from the dorsal compartment to the midgut. The lumen of the oesophagus is occluded by an anterior and two lateral folds which run for the greater part of its length. The oesophagus is constricted by contraction of a band of circular muscles round the oesophagus pressing the folds

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together. Since the folds are filled with loose connective tissue their arrangement permits maximal distension of the oesophagus during food ingestion. Such an arrangement is necessary since the oesophagus is lined by relatively inelastic cuticle. The cuticle (Fig. 3) is about 10 p thick and the underlying epidermis consists of densely staining columnar cells.

Fig. 2.-Transverse section of the anterior chamber of the proventriculus through the circular muscle layer; AC, anterior oesophagus (Fig. 1, section a-a). M,Muscles; CM, chamber; OES, lumen of oesophagus; CON,circumoesophageal connectives; MD, mandible; EP, epistome, containing glands.

Longitudinal folds also run along the sides of the anterior chamber of the proventriculus permitting expansion of the chamber if it is filled with food. Two pairs of longitudinal ridges in the floor of the chamber form channels which lead back to the digestive gland openings in the posterior chamber. The most median pair of ridges form a single small median channel in the anterior part of the chamber, but posteriorly these ridges become confluent and the channel is obliterated. The lateral ridges adjacent to the median ridges form open channels anteriorly, but project inwards posteriorly so as to enclose the median ridges, and become continuous with the ridges dividing the posterior chamber into dorsal and ventral compartments.

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These longitudinal ventral channels running forward from the digestive gland probably carry enzymes to the food in the anterior chamber (Yonge 1924; Pike 1947). (4 (b)

Fig. 3.-(a) Lining of oesophagus. (b) Lining of anterior chamber of proventriculus. E, Epicuticle; P, procuticle; ED, epidermis; c, connective tissue; M, muscle.

The cuticle of the anterior chamber (Fig. 3) is about 5 p, the epicuticle being 1-2 p thick. As in the oesophagus, the epidermis is a layer of densely staining columnar cells, and contrasts with the attenuated epidermis of the external body cuticle during the intermoult period (Dall 19653). This is further discussed in Section III(b) below.

Fig. 4.-Transverse section through the posterior chamber of the proventriculus (Fig. 1, section b-b). DC, Dorsal compartment; c, connective tissue; M, muscle; vc, ventral compartment; LG, longitudinal groove between rows of setae.

In the posterior chamber the lateral ridges with their dense setae permit only the finest particles to pass into the ventral compartment. The large ventral median

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ridge of the latter bears longitudinal rows of dense setae with grooves between them where the fine food particles collect, and are then passed backwards to the openings of the digestive gland (Fig. 5). Normally the dorsal compartment is more or less Bled with food particles of various sizes, but in the ventral compartment food is restricted to the longitudinal grooves.

Fig. 5.-Transverse section through the openings of the digestive gland into the posterior chamber of the proventriculus (Fig. 1, section c-c). LM, Lappets which extend backwards into the midgut; DC, dorsal compartment; vc, ventral compartment; OD, opening into digestive gland; s, secondary channel into which digestive gland tubules open; TO, tubule opening adjacent to proventriculus openings.

(ii) Digestive Gland (Figs. 1 and 6) The general structure of this organ agrees with the descriptions of Yonge (1924), van Wee1 (1955), and Pillai (1960). The bulk of the tissue comprises simple tubules whose walls consist of a single layer of secretory epithelium. Only a few scattered cells and small blood vessels occur between individual tubules. Some of the tubules open directly into the region adjacent to the openings of the proventriculus, but others are grouped round secondary channels leading from this region (Fig. 5). The distal ends of the more dorsal tubules meet above the posterior proventriculus,

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while the more ventral tubules meet below it. The tubules are connected by sparse connective tissue, and the whole organ is invested in a yellow-brown membrane with guanine-like deposits scattered over its surface. Thus the digestive gland completely surrounds the posterior proventriculus. The apex of each tubule consists of a cuboidal epithelium of closely packed cells which stain intensely and which exhibit metachromasy with P.A.S. and toluidine blue. Immediately below this area is a zone of transitional cells with similar staining characteristics, but which are columnar. The remainder of the tubule consists of columnar cells with extensive cytoplasm. The upper part of this region is shown in Figure 6. Two major cell types appear to be present. The most abundant form vacuoles (Fig. 6, vc) which become progressively larger, and ultimately fill and distend the cell. Detached cells were not observed in properly fixed tissues, and secretion therefore appears to be merocrine. The other cell type (Fig. 6, MC) has a dense cytoplasm and large nucleus, and exhibits metachromasy with P.A.S. and toluidine blue. These cells apparently contain a large amount of mucopolysaccharide (Dall 1965~).No cells were observed showing graded intensities of metachromasy, nor was there any other evidence to support van Weel's (1955) and Pillai's (1960) concept that these are merely a stage in a secretory cycle. Miyawake, Matsuzaki, and Sasaki (1961) report mucopolysaccharide-containing cells which also contain calcium, the amount of the latter fluctuating with the moult cycle. Thus there appears evidence for two distinct cell categories, a secretory and a "storage" type, as described by earlier authors (see Vonk 1960). Davis and Burnett (1964) present evidence that in the crayfish all tubule cells arise from the embryonic cells of the apex and go through absorptive, then secretory and fibrillar stages, and finally atrophy. The apical region of the tubule is adjacent to the haemocoel, and absorption from this region seems likely. However, holocrine secretion has been described in crayfish (Hirsch and Jacobs 1929, 1930), and atrophy of cells may not be a general feature of Decapoda. No such cells could be distinguished in Metapenaeus. In addition, the digestive gland is generally believed to play an important role in storage in preparation for moulting (Drach 1939; Renaud 1949). Apart from a few scattered interstitial cells, tubule cells comprise almost all of the digestive gland, and any such storage functions must has reside in these cells. However, Dall (1965~) shown that the "storage" cells are abundant immediately after moulting and suggested that they may secrete mucins involved in digestion. Hence the role of particular cells in storage of substances utilized for moulting is not clear. (iii) Midgut (Figs. 7 and 8) Though morphologically part of the midgut, the digestive gland is functionally closely associated with the proventriculus and has therefore been considered separately. In crayfish and some other Decapoda the proctodoeal cuticle extends almost to the openings of the digestive gland and the digestive gland is virtually the only portion of the digestive tract obviously derived from the embryonic midgut (Balss 1927). The latter author remarks that the midgut varies considerably in length throughout the Malacostraca. Yonge (1924) found that the midgut of Nephvops extended for the greater part of the length of the abdomen, and in Metapenaeus the midgut reaches to the sixth abdominal somite.

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Fig. 6.-Transverse section of a digestive gland tubule, below the transitional region (see text). vs, Secretory cells forming vacuoles of digestive enyzmes (these vacuoles ultimately coalesce and distend the cell); MC, cells containing mucopolysaccharide; BB, "brush-border"; AT, adjacent tubules separated only by a thin layer of connective tissue.

Fig. 7.-Transverse section through the anterior midgut (Fig. 1, section e-e). PM, Peritrophic membrane containing faeces; CE, columnar epithelium; LML, longitudinal muscle layer; a circular muscle layer lies immediately inside this; a thin connective tissue covers the outer muscle layer.

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The midgut is lined throughout its length by dense columnar epithelium which is surrounded by muscular and connective tissue (Fig. 7). No "vermiform bodies", described as occurring in the posterior midgut of Caridina (Pillai 1960), could be found. A peritrophic membrane is secreted by the anterior midgut. The epithelium extends into paired anterior diverticula (Fig. 8) and a single posterior diverticulum (Fig. 9). These diverticula open at the terminations of the stomodoeal and of the proctodoeal cuticle, respectively, and appear to be simple extensions of the midgut.

Fig. 8.-Transverse section through the openings of the anterior diverticula of the midgut (Fig. 1, section d-d). AL, Lumen of a diverticulum (the two lumina fuse above and below the gut); CUT, cuticle which lines the canal; CAN, canal leading from the foregut (G) to the lumen of the diverticulum; CE, columnar epithelium; LD, lappet which occludes the opening to the diverticulum. (The cuticular lining of the gut stops just behind the openings to the diverticula.)

The openings of the anterior diverticula are covered by a pair of lappets extending backwards from the proventriculus. Food particles were never found in the diverticula in sections from many animals, although food was always present in the midgut, and the lappets with their fringing setae therefore appear to function as valves. Particulate matter was also invariably absent from the posterior diverticulum, but this is accounted for by the presence of the peritrophic membrane in this region. Epithelial cells of the diverticula are taller and more closely packed, and stain more intensely than those of the midgut, but otherwise the epithelia are similar. There is every appearance that the epithelium of the diverticula is very active, and it is unlikely that the structures are vestigial. It is possible that the diverticula have a

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secretory function, and may play a part in excretion of calcium and other salts via the anus (Dall 1965d, and unpublished data). However, the midgut lining also has the appearance of a secretory epithelium. In both diverticula and midgut, vesicles form at the bases of the epithelial cells, and seem to be extruded into the lumen. The vesicles contain refractile, non-birefringent, granular, acidophilic material, which does not stain metachromatically with P.A.S. or with toluidine blue; these structures appear to be similar to Pillai's (1960) "cytoplasmic bodies". No definite function as yet can be assigned to these vesicles, but evidence is growing to support the longestablished view that the midgut has an excretory function (see Parry 1960).

Fig. 9.-Lateral dissection of the sixth abdominal somite, showing posterior midgut (MP)posterior diverticulum (D) rectum (R), and anus (A). T, Telson; RP, one of the six longitudinal rectal pads (four are shown); NC, nerve cord; u, uropod.

(iv) Rectum (Fig. 9) At the junction of the midgut and rectum the epithelium of the former ends abruptly, and the six large pad-like rectal ridges begin. These ridges are filled with a spongy reticular tissue which did not take up any of the stains used. Posteriorly the ridges taper down to longitudinal muscle bands. Circular muscles surround the rectum, and their contraction presses the pads together and so may close the rectum. The role of the pads and their longitudinal muscles is discussed below.
(b) Functions o the Digestive Tract in Feeding f

(i) Food Ingestion The animal normally appears to browse selectively on surface particles of organic detritus. The three pairs of chelate appendages bear tufts of setae which apparently have both taste and tactile functions. For example, when the animal encounters a larger piece of food, the chelae rapidly explore the surface before the food mass is seized. During browsing, the chelae appear to search systematically over the surface of the substratum, and presumably gather clusters of micro-organ-

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isms and other edible matter, which are then conveyed to the mouthparts. That this feeding is selective is indicated by the very small quantity of inorganic particles in the gut. Larger food masses are eaten while being held between the external maxillipeds. If the food is tough (e.g. muscular tissue) the maxillipeds assist in detaching pieces by pushing the food mass away while a portion is grasped by the mandibles. Feeding ceases when the anterior chamber of the proventriculus is filled, although large food pieces may be held by the maxillipeds until feeding is resumed. A starved animal is able to fill the anterior proventriculus with soft food (e.g. cooked liver-cereal homogenate) in less than a minute. (ii) Permeability of the Anterior Proventriculus Yonge (1936) found that the proventriculus of Nephrops was permeable to glucose. Dall (1965~) observed that [14C]glucose introduced into the anterior proventriculus was metabolized very rapidly. The thin yticle of this structure with its apparently active epithelium, noted above, raises the question whether there may be significant absorption through the proventricular wall. Proventriculi were dissected out of freshly killed animals, large muscles were removed, and the organ was flushed out with iso-osmotic sea water containing 22Na or [14C]glucose. A ligature was then tied just behind the gastric mill, the anterior chamber filled to capacity with labelled water, and the oesophagus ligatured. The preparation was washed for 5 min in three changes of iso-osmotic unlabelled sea water, and then suspended by a thread in similar water and stirred continuously. For 22Na counting the proventriculus was removed, washed once and counted in a well-type sodium iodide scintillation crystal. 14C was counted by transferring an aliquot to a planchet, drying and counting with a methacrylate fluor. Results are shown in Figure 10 where mean values of three experiments with each isotope are shown. 22Navaried at 5 hr from 75 to 90 %, while [X!] glucose varied from 6 to 12 % at 2 hr. The latter experiments were not continued beyond 2 hr to avoid possible effects from bacterial decomposition. Permeability to sodium is high, but, results indicate that permeability to glucose is insufficient to permit a significant amount of digested carbohydrate to be absorbed in this way. (iii) Rate of Food-mass Movement Animals were fed cooked, homogenized liver-cereal into which finely divided llOAg particles had been ground, the mixture being rolled into pellets. Shrimp were washed after feeding, placed in plastic gauze tubes, and regions of the gut counted by positioning the animal over an 8-mm gap between two 50-mm thick lead bricks, with a scintillation counter below the gap. This method obviously does not permit discrimination between adjacent regions, but it was found that passage of food through anterior proventriculus, digestive gland region, anterior and posterior midgut could be detected. Also, there is no measure of digestion since only inert particles are being traced. In all, three batches of three animals each were measwed. In one batch the shrimp were placed in the tubes only for counting, whereas in the other two they were left undisturbed for the duration of the experiment. No appreciable differences could be detected.

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Food begins to leave the anterior proventriculus almost as soon as it is flled, by 2-3 hr there is a marked decline, and by 6-12 hr the chamber is usually empty. The digestive gland area reaches a maximum value 0.5-1 hr after feeding and declines slowly to zero over a 12-20-hr period. In some cases there was an abrupt drop in radioactivity indicating that there is a mechanism for emptying the blind tubules of the digestive gland of indigestible particles. In starved animals some defecation may occur from 1 hr after feeding, but usually reaches a peak at 5-8 hr. However, the gut is not completely emptied of llOAg until 12-20 hr after feeding. A large part of digestion would therefore appear to be complete by about 8 hr, but the total process may take longer. The upper value of 20 hr is probably maximal and could represent only the final emptying of indigestible particles.

TIME (HR)

Fig. 10.-Diffusion of 22Na (@) and [W]glucose (+) through the anterior proventriculus, shown as percentage of radio-isotope in proventriculus at zero time.

(iv) Mechanims of Food-mass Movement Musculature and operation of the proventriculus is described by Young (1959) for Penaeus setiferus, and Metapenaeus bennettae appears to be similar in all essential features. Filling and emptying of the digestive gland must be achieved by the action of the proventricular muscles and the muscles surrounding the gland, since it has virtually no intrinsic muscle fibres, nor are there any muscle attachments on the surface. The complex movements of the proventriculus may be seen through the dorsal carapace, and the proventriculus is pushed rhythmically into the digestive gland. Very fine food particles may be observed in the proximal portions of the tubules, and these could be forced in by the action of the "presses" of the posterior proventriculus.

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Larger particles excluded from the digestive gland and residue from the latter are guided well back into the peritrophic membrane of the midgut by the backwardprojecting lappets of the proventriculus. These form a U-shaped channel much as in Nephrops (Yonge 1924). Strong peristaltic contractions of the isolated midgut may be observed in vitro. These contractions probably assist in moving the peritrophic membrane along in vivo. The rectal pads appear to play an essential part in defecation. During this process, the rectum contracts a number of times in rapid succession, and a length of intact peritrophic membrane containing faeces, approximately the length of the midgut, is pushed out. The rectal pads apparently serve to grip the peritrophic membrane and as the rectum contracts so a section is extruded. Thus the animal produces characteristic long faecal pellets. IV. DISCUSSION The proventriculus of Metapenaeus appears to function in essentially the same way as that of Nephrops (Yonge 1924), Astacus (Balss 1927), and GaIathea (Pike 1947). Yonge (1936) found that the proventriculus of Nephrops was fairly permeable to sodium chloride, but that glucose penetrated "extremely slowly". Results with Metapenaeus show that sodium equilibrium would be reached in 6-7 hr. Dall (1965d) found that abdominal cuticle was permeable to 45Ca; the high permeability of the proventricular cuticle to 22Na is therefore to be expected. Glucose appears to diffuse at about one-seventh the rate of 22Na,and although this is an appreciable rate, glucose absorption is probably a negligible factor in normal absorption of digested food. Dall (1965~)found that [14C]glucose introduced into the proventriculus was rapidly metabolized, and suggested that the glucose may have been absorbed directly from the proventriculus. It is more likely that the glucose was passed rapidly to the digestive gland and absorbed there, since, in the experiments described above, labelled food was was detected in the digestive gland a few minutes after feeding. As noted above, filling and emptying of the digestive gland must be due to the action of the proventriculus and surrounding thoracic muscles. Yonge (1924) described circular and longitudinal muscle fibres between the digestive gland tubules in Nephrops, but Pearson (1908), Pike (1947), and Pillai (1960) indicated that sparse connective tissue and blood vessels were the only interstitial tissues in the animals they examined. Sections of Metapenaeus stained with Mallory's stain revealed a few fibrous strands between the tubules and these gave a typical collagen blue-staining reaction. Occasional nuclei were scattered along the fibres. Phase-contrast examination did not reveal muscle fibres. Nephrops appears to be unique in the possession of intrinsic digestive gland muscle fibres, and other species so far described appear to need some extrinsic mechanism for filling and emptying the gland. Fox (1952) recorded oral drinking in a number of Decapoda, and a large water intake has been found to occur in Metapenaeus (Dall, unpublished data). This water, plus muscular action, could play an important role in filling and emptying the digestive gland. In addition to the digestive gland, gut diverticula occur in a number of Decapoda (Calman 1908; Pearson 1908; Balss 1927 ; Young 1959; Pillai l960), but so far no definite function has been revealed. They are absent in the freshwater Astacus (Balss

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1927), and functions associated with regulation in the marine environment might be postulated, were it not for the fact that they are absent in Galathea (Pike 1947). The diverticula are particularly well developed in Cancer (Pearson 1908), and the present author has found a similar development in several species of Portunidae, Grapsidae, and Ocypodidae, and it is likely that the Brachyura are all similar in this respect. No diverticula could be detected in the brackish-water shrimp Macrobrachiurn equidens (Palaemonidae), but another caridean freshwater shrimp Caridina laevis (Atyidae) has three (Pillai 1960). Young (1959) described midgut diverticula in Penaeus setiferus, and the writer has found them in Penaeus esculentus and P. plebejus as well as in Metapenaeus bennettae. A number of penaeid species are euryhaline (MacFarland and Lee 1963; Dall 1964), and Metapenaeus bennettae extends into regions where Macrobrachiurn occurs. It does not therefore appear likely that, in shrimp at least, the midgut diverticula are organs associated with osmotic regulation. Secretion of digestive enzymes is possibly a function, but this would be unlikely in the Brachyura, for in these the gut is lined with chitin almost from anus to digestive gland openings. A possible role in salt excretion has been mentioned above and will be the subject of a future paper. The midgut appears to be similar to that of the Norway lobster, Nephrops, (Yonge 1924), rather than to that of the shrimp Caridina, (Pillai 1960). A peritrophic membrane has been recorded in various Caridea (Forster 1953), and is probably characteristic of those Malacostraca with a long midgut, where it is a functional necessity. Rectal pads have been described in Caridina (Pillai 1960), but unlike Metapenaeus where the pads are quite smooth, spinules are present. Macrobrachium equidens has rectal pads similar to those of Metapenaeus. As well as its role in expelling faeces in the peritrophic membrane, the rather elaborate rectum also appears to have an important function in anal swallowing of water. Fox (1952) described this in a number of Crustacea, Pillai (1960) made some detailed observations of rectal drinking in Caridina, and Dall (1965d) found evidence of uptake of calcium via the anus. Rectal "pumping" was observed in both Metapenaeus and Macrobrachiurn when no peritrophic membrane was being extruded and water intake appeared to be taking place. To what extent this serves to assist in defecation and gut distension as proposed by Fox (1952), or whether it serves some other function, remains to be determined.

The author's thanks are due to Miss C. Dunning for her assistance in preparing many of the slides used in this work, which was made possible by a grant from the Commonwealth Research Grants Committee. VI. REFERENCES
BALSS,H. (1927).-In "Handbuch der Zoologic". (Ed. Dr. Willy Kukenthal.) Vol. 3, Pt. 1. (Walter de Gruyter and Co. : Berlin.) CALMAN, T. (1909).-In "A Treatise on Zoology". (Ed. Sir Ray Lankester.) Pt. 7, AppendiW. culata; Fascicle 3, Crustacea. (Black: London.)

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