The effect of starvation on the metabolic rate and ...

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this species is not present in soil (Hubert et al., 1999; ... same conditions, but pieces of bark covered with the green bark alga, Desmococcus vulgaris (syn.
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(4) Activity of digestive enzymes. Amylolytic (EC 3.2.1.1) activity was assayed after 21 day. Whole-body homogenates of 64 living mites were used, with 12 replicates per group. The mites were weighed and homogenised in 2.5 ml of phosphate (Britton-Robinson) buffer (pH 7) and centrifuged (6000 rpm, 7 min). The pH optimum of Galumna elimata amylase was 7 (Šustr, unpublished). The S-Test using specific chromolytic substrates (Institute of Chemistry Bratislava, Slovakia) was used for the amylase assays (see Šustr & Hubert 1999). The catalytic activities of the enzymes were expressed in mg of decomposed substrate per hour per 1 g of fresh oribatid body mass (mg. h-1. g-1). The significance of the difference between experimental groups was tested using nonparametric Kruskal-Wallis test (Statgraphics®). (5) Microanatomical observations. Living individuals from both groups were fixed in modified Bouin-Dubosque-Brasil fluid (Smr , 1989) after 21 and 42 days. The fixed mites were embedded in paraplast, sectioned (thickness 5–7 µm), and stained in Masson's triple stain. RESULTS

The mortality of starved mites was significantly higher than of the control mites after 21 and 42 days (log-linear analysis, F2 = 3223, P < 0.001) and mortality was higher after 42 days than after 21 days (log-linear analysis F2 = 1909, P < 0.001, Tab. 1). Galumna elimata mites in the control group fed intensively on the green bark algae, Desmococcus vulgaris. Juveniles were observed after 42 days in the control group, but not in the starved group. Starvation significantly influenced the mass of individuals after 21 days. The mean fresh mass of fed individuals was 121 µg, dry mass 57 µg, and the water content 64 µg (53 % of the fresh mass). The mean fresh mass of starved mites was lower (113 µg). Their dry mass was 41 µg and the water content about 63 % (Fig.1). Water content was significantly (P = 0.03) higher in the starved mites. The respiration rate was significantly lower in the starved than in fed mites. The respiration rate was 240 µl.g-1.h-1 in the fed individuals and 130 µl.g-1.h-1 (Fig. 2) after 21 days of starvation. Starvation for 21 days did not influence the amylolytic activity. Mass specific amylolytic activity was 5 903 mg-1.g-1.h-1 and 5 762 mg-1.g-1.h-1 in the control and starved group, respectively (Fig. 3). The digestive tract of Galumna elimata (Figs. 4, 5) is similar to the digestive tract of the model species Ceratozetes cisalpinus (see Woodring & Cook, 1962b) and Euzetes globulus (see Hoebel-Mävers, 1967).

Fig. 1. Difference after 21 days in the mass of starved and control Galumna elimata. DW dry mass, Water mass of body water, S starved group, C control group. The numbers in column = number of replicates.

All parts of the gut of the control mites contained algal food boli (Figs. 4, 5, 7). The guts of the starved mites were full of mucoid substances (Fig. 8) or empty (Fig. 6). After 42 days of starvation their tissues appeared reduced (Tab. 2, Fig. 6). Some fluid substances present in the salivary glands (Fig. 19), and mucoid substances in the caeca (Fig. 5) and by the mesenteral cells (Fig. 10). These substances were mixed with the ingested algal cells in fed individuals, or concentrated in the middle of mesenteron in starved individuals. In starved animals, the mucoid substances filled the whole mesenteron (Fig. 8) and more concentrated mucoid droplets formed boli (Fig. 9). The boli were passed through the gut into the rectum (Fig. 18). There were no structural differences in the mesenteral and faecal boli. After 21 days of starvation, individuals with mucoid boli prevailed over individuals lacking mucoid droplets in the mesenteron. Individuals lacking mucoid substances in the mesentron prevailed after 42 days. Generally, mesenteral cells were thicker in the anterior than in the posterior part of the mesenteron of the control

TABLE 1. Mortality of starved and control Galumna elimata Period Number of individuals that: Control group Starved group Analysis of deviance Null Groups Periods

21 days

42 days

survived 99 85

died 2 19

survived 67 59

died 12 45

Df.

Residual deviance

1 1

3,539.7 1,708.8

Df. (res. Dev.) 3 2 1

Deviance 5,396.6 1,856.8 148.1

267

Fig. 2. Difference after 21 days in respiration of starved and control Galumna elimata. M/W mass specific oxygen consumption, S starved group, C control group, the numbers in column = number of replicates.

Fig. 3. A comparison after 21 days of the amylolytic activity in starved and control Galumna elimata. EA mass specific amylolytic activity, S starved group, C control group.

mites. The cells were filled with dark stained granules (Fig. 10). There were green stained microvilli on the apical parts of the cells. The nuclei were relative large and well stained. These cells produced little apocrine secretion compared to the control specimens, there were fever granulae in the cells, and the thickness of mesenteral cells was not reduced after 21 days of starvation (Fig. 11). After 42 days, the mesenteral cells were very thin, and the nuclei still lacked dark stained granulae. No microvilli were observed (Fig. 12). The caecal cells in control individuals exhibited intense apocrine secretion (Fig. 13). The cells were strongly vacuolized and produced mucoid substances and large green stained spherical granulae. The caeca of mites starved for 21 days were similar to those of the control group, only lacking green granulae (Fig. 14). After 42 days of starvation, the cells of caeca were thinner and poorly vacuolized and lacked apocrine secretion (Fig. 18). The cells of the colon (Fig. 8) were similar to those of the model species (see Woodring & Cook, 1962b; Hoebel-Mävers, 1967). The rectum of control individuals had a reduced brush border (Fig. 16), while the rectum of animals starved for 21 days had a well-developed brush border (Fig. 17). The

thickness of the colon and rectal cells, as well as their brush border were reduced after 42 days of starvation (Fig. 18). The proventricular glands (for description see Šustr & Hubert, 1999) were present in the majority of specimens from the control group and in some animals starved for 21 days, but apparently absent after 42 days of starvation. The salivary glands were lobed. Some cells contained a large nucleus and vacuoles. Other cells in the lobes formed reservoirs. There were two kinds (physiological types) of salivary glands in control individuals; namely those without reservoirs with vacuolized cells (Fig. 19) and glands with well developed reservoirs. The second type prevailed in the starved mites (Fig 20). The mesenchymal cells of control specimens contained glycogeneous granulae (Fig. 15). These glycogeneous granulae were not observed in starved mites, except for one gravid female that had been starved for 21 days. Extra-intestinal bacteria were observed in one specimen starved for 21 days (Fig. 21). The male/female ratio tended to 1:1, however females prevailed over males after 42 days of starvation. Testes and seminal vesicles were not different from those of the model species Ceratozetes cisalpinus (see Woodring &

Figs 4–12. 4 – Galumna elimata, control group - sagital section; 5 – horizontal section; 6 – horizontal section of an individual starved for 42 days; 7 – intensive consumption of algae by control individual; 8 – mesenteron of an individual starved for 21 days filled with mucoid substances of; 9 – mucoid bolus in an individual starved for 21 days; 10 – mesenteron of a control individual, the arrows point to mucoid package of food; 11– apocrine secretion in the mesenteron of an individual starved for 21 days; 12 – mesenteron of an individual starved for 42 days. Abbreviations used: c – colon, ca – caecum, cm – cheliceral muscles, cu – cuticle, e – egg, fb – food bolus, m – mesenteron, mc – mesenteral cell, md – mucoid droplets, ms – mucoid substances, mu – muscles, o – oesophagus, ph – pharynx, r – rectum, sg – salivary glands, syn – synganglion. Scales: 0.1 mm … 4–6; 0.5 mm …7–9; 0.025 mm…10–12.

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