Sphaerospora epinepheli N. Sp. (Myxosporea: Sphaerosporidae) Observed in Grouper (Epinephelus malabaricus)

Sphaerospora epinepheli n. sp. is described from grouper, Epinephelus malabaricus , in cage-cultured and wild fish collected from both coastal lines of southern Thailand. Subspherical to spherical spores and mono- or disporous pseudoplasmodia were observed in the lumen of kidney tubules. Pseudoplasmodia were round to elongate, size range 15.6–22.9 μm (length) × 8.4–21.6 μm (width). Spores were 7.8–10.0 μm (length) × 12.3–14.5 μm (thickness), and 7.0–9.5 μm (width) with two spherical polar capsules of equal size measuring 2.9–4.4 μm in diameter and containing polar filaments with six or seven windings. Two uninucleate sporoplasms showed iodine vacuoles. Blood stages, similar to C-blood protozoans observed from freshwater fish in Europe, were found from peripheral blood smears of grouper. Ultrastructural studies of blood stages showed a similar structure to unidentified mobile protozoans from the blood of carp. Electron dense bodies were observed in the cytoplasm of the primary cell blood stages. Infected proximal-tubular epithelial cells showed highly vacuolated cytoplasm and pycnotic nuclei.     

Coccidia from the Tiger Salamander, Ambystoma tigrinum, in Northeastern Colorado and in Northern New Mexico

SYNOPSIS. Oocysts of Eimeria ambystomae Saxe, 1955, Eimeria microcapi sp. n., and Eimeria urodela sp. n. are described from the tiger salamander, Ambystoma tigrinum , collected in Colorado and New Mexico. The oocysts of E. ambystomae are ellipsoid, 29.8 × 17.3 (24–38 × 15–25) μm, and the sporocysts lanceolate, 22.6 × 5.4 (16–27 × 5–7) μm. Oocyst and sporocyst residua are present, but not a polar granule and a micropyle. The oocysts and sporocysts of E. microcapi are ellipsoid, measuring respectively 38.1 × 25.3 (35-41 × 23-26) μm and 18.1 × 7.4 (16-19 × 6–8) μm. Oocyst and sporocyst residua, a micropyle (mean 3 μm), and a distinct micropyle cap (2 μm high) are present, but not a polar granule. The oocysts of E. urodela are spheroid, 22.2 (14-26) μm, and the sporocysts lanceolate, 16.3 × 5.8 (12-19 × 4-7) μm. Oocyst and sporocyst residua are present, but a polar granule and a micropyle are absent.     

Some effects of low pH and calcium on the growth and tissue mineral content of yearling brown trout, Salmo trutta

Yearling brown trout, Salmo trutta , were exposed to low mineral content water (nominal concentrations of 20μmol 1⁻¹ magnesium, 7.7 μmol 1⁻¹ potassium, 44 μmol 1⁻¹ sodium) over a pH range of 4.0–5.2 with ambient calcium concentrations of 2.5–60 μmol 1⁻¹. All fish died at pH 4.0 and 4.2 irrespective of ambient calcium concentration and also at pH 4.4 with only 2–3 μmol 1 ⁻¹ calcium (that is calcium-free water except for that leached from the diet or excreted by the fish). Good growth rates were obtained over the remaining treatments which extended down to pH 4.4 with as little as 7 μmol 1⁻¹ calcium. When starved, weight loss was inversely correlated with pH. Effects on plasma chloride, percentage dry weight and calcium, potassium sodium, and phosphorus contents of skin, muscle and bone tissue were also investigated. These demonstrated pH effects on mineral metabolism in starved fish, but no effects were detected in fed fish.     

Henneguya adherens N. Sp. (Myxozoa, Myxosporea), Parasite of the Amazonian Fish, Acestrorhynchus falcatus

ABSTRACT. A new species of myxosporean from the gill filaments of the freshwater teleost fish, Acestrorhynchus falcatus collected in the Amazon river is described from light and transmission electron microscope observations. The mature spores (total length 32.3 [30.7–35.1] μ) and all developmental stages were found in the same sporogonic plasmodium. The ellipsoidal spore body consists of 2 unequal shell valves adhering together along the suture lines. Each valve, tapering as a caudal projection, forms a long tail (length 20.5 [18.0–21.7] μm). The tail was surrounded by a homogeneous sheath on its length. The polar capsules measuring 3.1 × 1.2 μm contain 3–4 coils of the polar filament. All surfaces of the immature and mature spores were surrounded by a closely adherent homogenous structural sheath, mainly thicker around the tails. The taxonomic affinities of this parasite to other species are discussed.     

Distribution of endosulfan in water, sediment and fish from Warri river, Niger delta, Nigeria

This article presents the first attempt to quantify the levels and distribution pattern of endosulfan, an organochlorine pesticide in surface water, sediment and fish ( Chrysichthys furcatus and Tilapia zilli ). The samples were collected from three stations (Ovwian, Ekakpamre and Ovu) of Warri River in the western Niger Delta of Nigeria in 2006 during the dry and wet seasons (January–August). A total of 96 samples made up of 24 samples each for water, sediment and fish were analysed in this study. The pesticide levels were analysed using high performance liquid chromatography (HPLC model CECIL 1010) to elucidate its distribution in various environmental compartments. The ranges of concentrations of the pesticide in the matrices were: 0.01–9.23 μg/l (water), 0.06–11.98 μg/gdw (sediment), 0.01–15.06 μg/gdw ( Chrysichthys furcatus) and 0.01–1.80 μg/gdw ( Tilapia zilli ). From this result, decreasing order of occurrence of the pesticide is as follows: fish > sediment > water. The concentrations observed in fish ( Chrysichthys furcatus ) were higher than the levels observed in sediment and water suggesting bioaccumulation of the pesticide by the fish. Spatial variations occurred with downstream stations having statistically higher concentrations in all matrices at P  < 0.05. Seasonal variations occurred with higher concentrations in dry season for water and sediment only, while the fish species had higher concentrations in the wet season. The observed values of endosulfan were above the ecological bench marks (0.02 μg/l) recommended by Nigeria Environmental Protection Agency and European Union. They were also relatively higher than those in previous studies on the Nigerian environment, an observation that calls for regular monitoring of the Niger Delta water bodies.     

Induced spawning by LHRHa and pimozide in the Asian catfish Clarias macrocephalus (Gunther)

Experiments were conducted to determine the optimum dose of luteinizing hormone-releasing hormone analogue (LHRHa) and pimozide (PIM) injected simultaneously to yield a high ovulation rate and produce sufficient eggs in the Asian catfish Clarias macrocephalus . In June 1990, injection of 0.05 or 0.10 μg LHRHa/g body weight (BW) + 1 μg PIM/g resulted in 100% ovulation, while only 80% of gravid catfish injected 0.025 μg LHRHa + 1 μg PIM/g ovulated. Most of the eggs stripped from 6 out of 8 control fish were not mature. Fertilization and hatching rates of LHRHa + PIM-induced fish (75–90% and 39–51%, respectively) were higher than those of control fish (36–39% and 0–1% respectively). In August and September 1990, at gravid catfish ovulated after injection of 0.05–0.10 μg LHRHa + 1 μg PIM/g BW. However, only 20% of the fish given 0.025 μg LHRHa/g + 1 μg PIM/g BW in August ovulated. No eggs could be striped from any of the control fish in August and September 1990. Techniques developed in this study, showed a simple and effective way of spawning captive catfish, C. macrocephalus . A simultaneous intramuscular injection of 0.05 μg LHRHa + 1 μg PIM/g and stripping of eggs at 16–20 h post-injection have been tested to yield high ovulation, fertilization and hatching rates.     

Oral administration of cholecystokinin receptor antagonists increase feed intake in rainbow trout

Individual immature rainbow trout consumed 1–2% body weight per day, but significantly more ( P < 0·001) when fed by hand than by demand feeder. When treated with CCK antagonists (L 364, 718; 100 μg kg⁻¹ on day 12 or SR 27, 897; 50 μg kg⁻¹ on day 16), the fish ate significantly more than their mean daily intake on the other days of the experiment. This increase in feed intake was affected by the feeding technique: hand-fed fish increased by 70–80% their feed intake while in demand-fed fish the increase was significantly less (50–60%). However, the increase in feed intake observed on days 12 and 16 was identical for both drugs used.     

Morphological Comparison of Diophrys scutum (Dujardin, 1841) and Diophrys peloetes n. sp. (Hypotrichida, Ciliophora)

SYNOPSIS. Diophrys scutum , collected from four locations on the New Hampshire coast, ranged from 89–195 7mu; in length, 50–105 μ in width, and 68–88 μ in buccal cavity length. The end of the adoral zone of membranelles (AZM) extends 37–59 μ (average = 45.6 μ) posteriorly in a groove on the right side of the body. Dorsally are five rows of stiff cilia. The silverline system (Chatton-Llvoff technique) appears as a fine meshwork, entirely different from that found in Euplotes or Uronychia . There are two elongate macronuclei (Feulgen reaction) and several micronuclei. Diophrys peloetes n. sp., collected from one location in Alligator Harbor. Florida, ranged from 95–134 μ in length, 62–84 μ in width. and 60–80 μ in buccal cavity length. The terminal portion of the AZM extends posteriorly in a groove 30–44 7mu; (average = 36.8 μ) on the right side of the body. Dorsally are eight rows of stiff cilia. The details of the silverline system are similar to those of D. scutum .
There is insufficient difference in ranges of body length, width. and buccal cavity length to use these characters in separating the two species. However, a statistical analysis shows that the length of the portion of the AZM on the right side of the body in D. scutum is significantly different (longer) from that of D. peloetes. Furthermore, these two species differ not only in number of dorsal ciliary rows, but also in the number of cilia per row. The degree of difference in these two species is similar to that between closely related species in other hypotrich genera, and also to that between some varieties of Paramecium aurelia.     

Mercury and chlorinated hydrocarbons in the food chain of Lake Päijänne, Finland

The sediments and various organisms in Lake Päijänne were examined for contaminants. The average mercury content of water plants was 9, of plankton 14, of sediment 114, of zoobenthic predators 83, of fish 332–1510 and of birds 240–13685 μg kg⁻¹ (wet weight). The average PCB content of plants was 3, of plankton 21, of the zoobenthos 44, of fish 36-117 and of birds 219–13490 μg kg⁻¹. The average ΔDDT content of plants was 0.5, of plankton 6, of the zoobenthos 14, of fish 7–42 and of birds 144-8262 μg kg⁻¹. Regional differences in mercury content were most pronounced in sediment and fish. PCB concentration was highest near a town. ΔDDT was quite evenly distributed. Water plant species did not differ from each other, nor did the plankton fractions. The zoobenthic predators contained more chlorinated hydrocarbons than did the herbivores. There were clear differences between most species of fish and the chlorinated hydrocarbon content was highest in vendace. In adult birds levels of all residues were significantly higher than in juveniles.
In most cases PCB content was positively correlated with ΔDDT and in birds PCB, ΔDDT and mercury levels were correlated. DDT residues occurred mostly as DDE, but in vendace the proportion of DDT was high. At most trophic levels, ΔDDT/PCB was 0.15-0.40 but in birds it reached 1–2.     

A Comparison of the Effects of Ethanol and the Competitive Glycine Antagonist 7-Chlorokynurenic Acid on N-Methyl-d-Aspartic Acid-Induced Neurotransmitter Release from Rat Hippocampal Slices

Abstract: N -Methyl- d -aspartate (NMDA; 500 μ M ) stimulated the net release of preloaded tritiated norepinephrine from rat hippocampal slices. Both ethanol and the competitive glycine antagonist 7-chlorokynurenic acid (7-CK) dose-dependently inhibited NMDA-stimulated release without affecting basal, nonstimulated efflux. These inhibitory effects were readily reversed upon washout of the drugs. Over the concentration range tested (25–200 m M ), ethanol inhibited ∼65% of NMDA-stimulated release with an estimated IC₅₀ of ∼70 m M . In contrast, 7-CK fully inhibited release (>95%) at a concentration of 30 μ M with half-maximal inhibition occurring at ∼2 μ M . The combination of 7-CK (1–30 μ M ) and ethanol (25–100 m M ) had an additive inhibitory effect on NMDA-stimulated release but did not alter the inhibitory potency of 7-CK. Calculated IC₅₀values for 7-CK in the presence of 25, 50, or 100 m M ethanol were (mean × SEM; μ M ) 2.33 (0.11), 2.38 (0.23), and 1.99 (0.30), respectively. 7-CK (3 μ M ) inhibited NMDA-stimulated [³H]norepinephrine release by ∼50%. This inhibition was fully attenuated by the addition of the glycine agonistserine with complete reversal occurring at 30 μ M d -serine. Increasing the 7-CK concentration to 10 μ M shifted the d -serine dose-effect curve to the right in a parallel fashion as expected for a competitive antagonist. In contrast, the inhibitory effects of ethanol or the combination of 7-CK (3 μ M ) and ethanol (25 or 50 m M ) were not reversed by the addition of d -serine (0.1–1,000 μ M ). Together, these results suggest that ethanol's inhibition of NMDA-stimulated [³H]norepinephrine release from hippocampal slices is not due to a simple competitive interaction with the glycine site on the NMDA receptor.     

Relationship of plasma steroids to germ cell development and the presence of protamine mRNA in rainbow trout during the induction of spermatogenesis with partially purified salmon gonadotropin

The gonadosomatic index (GSI) of pre–pubertal male rainbow trout, which had been injected biweekly with partially purified salmon gonadotropin (sG–G100, 50 μ mUg kg⁻¹ body weight), increased from 0.05 to 1.85 over 21 weeks from injection, while control GSI remained below 0.05. Plasma testosterone (T) increased from 2 to 11.34 ng ml⁻¹ by week 21 in injected fish, while control level remained below 1.5 ng ml⁻¹. In injected fish plasma 11–ketotestosterone (KT) and 17,20–dihydroxyprogesterone (17α20βP) levels increased from 20.2 to 41.9 and 8.9 to 219.7 ng ml⁻¹ respectively. Plasma T, 11–KT, and 17α20βP were all correlated with the GSI ( P <0–001) in injected fish. The most advanced stage of germ cells present in the control fish were spermatogonia. However, in injected fish spermatozoa were present by week 21. Eggs fertilized at this time with spermatozoa from injected fish achieved a 78% fertilization rate, whereas the testicular homogenate was incapable of fertilizing eggs.     

Time Sequence of Early Events in Fertilization in the Medaka Egg

The time sequence of early events in fertilization was examined in eggs of the medaka Oryzias latipes . The mean time after insemination required for sperm attachment to the egg surface through the micropyle depended on sperm concentrations. It was 3 ± 1 sec with a range from 1 to 6 sec after insemination when concentration of spermatozoa was high (about 2 × 10⁸/ml at 23°–25°C). The mean time from sperm attachment until cessation of its movement on the egg surface was 4 ± 1 sec with a range from 1 to 9 sec. Small cortical alveoli at the animal pole region within 15 μm of the sperm attachment point began to undergo exocytosis 9 ± 0.3 sec (range 5–16 sec) after sperm attachment. The velocity at which the exocytosis wave propagated increased from the earliest initiation point of exocytosis up to the 100 μm area, and became constant at about 12 μm/sec from 100 μm to 500 μm from the sperm attachment point. The present results suggest that at the time of fertilization in the fish egg, exocytosis of small cortical alveoli in the area about 15 μm away from the sperm attachment point occurs simultaneously.     

Susceptibility of Kenyan Wheat Varieties to Head Blight, Fungal Invasion and Deoxynivalenol Accumulation Inoculated with Fusarium graminearum

Fifteen wheat varieties commercially grown in Kenya were tested for their susceptibility to head blight and mycotoxin accumulation after inoculation with Fusarium graminearum in pot experiments. The strains of the pathogen used had been isolated from wheat collected in different growing areas of Kenya. Head blight susceptibility was assessed as the percentage of spikelets bleached and area under disease progress curve; kernel colonization by fungal mycelium was determined as ergosterol content. All varieties were found to be moderately to highly susceptible. However, the varieties differed in head blight susceptibility (29–68% of spikelets bleached; mean 54%), fungal colonization (67–187  μ g/g ergosterol content; mean 111  μ g/g) and the resulting mycotoxin contamination [deoxynivalenol (DON) 5–31  μ g/g; mean 13.5  μ g/g]. Grain weight reductions due to head blight ranged from 23 to 57% (mean 44%). The varieties could be therefore divided into partially resistant and highly susceptible genotypes. The kernels of highly susceptible varieties had higher mycotoxin and ergosterol contents. However, the kernels of some varieties contained more fungal mycelium (ergosterol) without the corresponding high amounts of DON, suggesting that they possess some resistance to DON accumulation. Less susceptible varieties showed resistance to fungal spread, as indicated by a slow disease development and lower content of fungal biomass.     

Life Cycle of Eimeria ferrisi Levine & Ivens, 1965 in the Mouse, Mus musculus

SYNOPSIS. The life cycle of Eimeria ferrisi is described from experimentally infected Mus musculus. The prepatent period was 3 days and the patent period was 3–4 days. The endogenous stages were found only in the cecum and colon. Three generations of schizonts were found. Mature 1st-generation schizonts first seen 24 hr postinoculation (PI) measured 10.9 (7–14) × 10.2 (6–13) μm and had 9.6 (7–14) merozoites. Some 2nd-generation schizonts had uninucleate merozoites and others had multinucleate merozoites. The former were first seen in small numbers 36 hr PI and were most abundant 48 hr PI. They measured 9.6 (5–13) × 7.9 (6–12) μm and had 18 (6–25) merozoites. Schizonts with multinucleate merozoites were seen 72 hr PI. Mature 3rd-generation schizonts were seen 72 hr PI. They measured 14.0 (12–18) × 11-0 (9–13) μm and had 12.5 (5–16) merozoites. Macrogamonts were first seen in 72 hr sections. Each young macrogamont had a large nucleus with a prominent nucleolus. Only one type of cytoplasmic granule appeared to be involved in the formation of the oocyst wall. Mature macrogamonts were 11.0 (5–14) × 10.0 (6–13) μm. Crescent-shaped bodies were observed in the parasitophorous vacuole of trophozoites and young macrogamonts. Early microgamonts were first recognized at 96 hr by the presence of darkly stained and irregularly shaped nuclei. Usually, mature microgametes were arranged in long, narrow whorls at the periphery of the microgamont or in whorls at the surface of 2–5 compartments.     

Light and Electron Microscopy of the Spore of Myxobolus heckelii n. sp. (Myxozoa), Parasite from the Brazilian Fish Centromochlus heckelii (Teleostei, Auchenipteridae)

ABSTRACT. A myxosporean parasitizing the gill filaments of the freshwater teleost fish Centromochlus heckelii collected in the Tocantins River (Lower Amazonian Region, Brazil) is described using light and electron microscopy. This parasite produces spherical to ellipsoidal cyst-like plasmodia up to 250 μm in diameter, with a thick wall strengthened by several stratified juxtaposed crossed collagen layers, whose thickness varies according to the number of the layers. Several compressed fibroblasts are observed among the collagen fibrils. Deposits of spherical dense material are scattered at the internal periphery of the cysts. Plasmodia and different developmental stages, including immature and mature spores, filled the central region of the cysts. The spore body is ellipsoidal in valvar view and biconvex in sutural view. It is formed by two equal-sized and symmetric valves measuring 12.7 μm long (12.2–13.1) ( n =50), 6.6 μm wide (6.3–6.9) ( n =25), and 4.0 μm (3.7–4.4) ( n =20) thick. A thin layer formed by fine and anastomosed microfibrils is observed at the spore surface. Two equal, elongated pyriform polar capsules measure 2.9 μm (2.7–3.3) × 1.7 μm (1.4–2.0) ( n =25), each containing four or five oblique polar filament coils. The binucleated sporoplasm contains numerous spherical sporoplasmosomes, glycogen particles, and a large vacuole with fine granular matrix. Based on the morphological and ultrastructural differences and specificity of the host, we describe this isolate as a new myxosporidian, Myxobolus heckelii n. sp. (Myxozoa, Myxosporea).     

Species of Anguilla as indicators of mercury in the coastal rivers and lakes of Victoria, Australia

Mercury concentrations in the axial muscle tissue of most (243) of the 254 Anguilla australis and most (20) of the 27 A. reinhardtii collected from 30 sites in coastal rivers and lakes in Victoria, Australia, during 1975–78 were well below the Australian statutory health limit (0.5 μg g⁻¹ wet weight). For A. australis the mean mercury concentration was 0.17 μg g⁻¹ (±0.16 s.d. , range 0.01–1.60 μg g⁻¹); for A. reinhardtii the values were 0.37 ± 0.23 μg g⁻¹ (range 0.12–1.10 μg g⁻¹). Statistical analyses showed that variation in mercury concentration due to total length accounted for only 13% of the total variation in A. australis and 2% in A. reinhardtii whereas locality accounted for 54 and 68%, respectively. Both species are thus considered suitable as indicators of mercury pollution.     

Fine Structure of the Envelope and Micropyles in the Eggs of the White Sturgeon, Acipenser transmontanus Richardson

The structure of the egg envelope and micropyles of the white sturgeon was examined with light and electron microscopy. The mature ovum is 3.5–4.0 mm in diameter and is covered by a thick envelope (50 μm) that consists of four distinct layers. The outermost layer, L4, is electron dense and amorphous. Interior to this is layer three (L3), containing numerous pores, or ductules. Layer two (L2) contains screwlike projections that anchor L3 and L4 to the egg. The innermost layer, L1, is closely apposed to the oolemma. Numerous micropyles (average 7) are restricted to a 100–200 μm region at the animal pole and penetrate the egg envelope. The outer opening of these rnicropyles measures 15 μm in diameter. The micropylar canal tapers twice, eventually terminating at the oolemma with an inner opening diameter of 1.2 μm. The micropyles of the white sturgeon egg appear more complex than micropyles in other fish eggs.     

The Sporulated Oocysts of Eimeria illinoisensis n. sp. and of Other Species of Eimeria of the Ox

SYNOPSIS. The sporulated oocysts of 12 species of Eimeria occurring in the ox Bos taurus in the United States are described and differentiated. They are E. alabamensis, E. auburnensis, E. bovis, E. brasiliensis, E. bukidnonensis, E. canadensis, E. cylindrica, E. ellipsoidalis, E. illinoisensis n. sp., E. subspherica, E. wyomingensis and E. zuernii. Two other species, not yet found in North America, which are recognized as valid are E. pellita and E. thianethi. The sporulated oocysts of E. illinoisensis n. sp. are ellipsoidal or slightly ovoid, 24–29 by 19–22 μ with a mean of 26.3 by 20.7 μ; their sporocysts are 13–16 by 6–7 μ with a mean of 15.3 by 6.5 μ. This species was found in 3 cattle from one farm in Illinois.     

Predicting the growth of age-0 yellow perch populations from measures of whole-lake productivity

SUMMARY. 1. The relationship was examined between four measures of lake productivity [total phosphorus (TP) and chlorophyll a (Chl a) concentrations, zooplankton density and biomass] and growth in length and weight of age-0 yellow perch in ten central Alberta lakes.
2. In these lakes, average summer TP and Chl a levels were in the range 11–51 and 1.4–19.5 μg1⁻¹, respectively. The interaction of TP and Chl a could explain 61% and 57% of the variance in total length and wet weight, respectively, of age-0 yellow perch sampled at the end of August.
3. The ability to predict first year fish growth from lake productivity is strongest at low levels of productivity (TP<35μgl⁻¹). In the lakes studied, fish community structure is more complex at high levels of productivity (TP>35μg1⁻¹), and more data on complex community level interactions seem necessary to predict fish growth in these systems.     

Normal ranges of some blood chemistry parameters in adult farmed Atlantic salmon, Salmo salar

Blood samples from healthy adult Atlantic salmon fed an optimal diet in net sea pens were collected at intervals from October to May. Haematological determinations and biochemical serum analyses were carried out on 20 fish in each of seven samples. The ranges of haemato-logical values for sample means were: haematocrit 44–49%, haemoglobin 8.9–10.4 g 100ml⁻¹, red blood cell count 0.85–1.10 × 10¹² l⁻¹, MCV 441–553 × 10⁻¹⁵ 1, MCH 94–106 × 10⁻⁶ g, MCHC 19.4–21.7 g 100ml⁻¹ and leucocrit 0.43–0.96%. The ranges of enzyme activities in serum, for sample means, were: alkaline phosphatase 647–988Ul⁻¹, aspartate aminotrans-ferase 202–351 Ul⁻¹ and alanine aminotransferase 4–8 Ul⁻¹. The ranges of the other parameters analyzed in serum were: total protein 41.6–56.6 gl⁻¹, albumin 18.3–24.3 gl⁻¹, albumin/total protein ratio 39.3–44.0%, creatinine 26–46 μmol, triglycerides 2.53–4.98 μmol and cholesterol 9.3–12.8 μmol. These values are considered to be the normal ranges in healthy fish. Variations due to seasonal changes, and the clinical significance of the selected parameters, are discussed. Data showing the reproducibility of the biochemical analyses in serum are presented.