Evaluating morphometric and metabolic markers of body condition in a small cetacean,the harbor porpoise (Phocoena phocoena)

Mammalian body condition is an important individual fitness metric as it affects both survival and reproductive success. The ability to accurately measure condition has key implications for predicting individual and population health, and therefore monitoring the population‐level effects of changing environments. No consensus currently exists on the best measure to quantitatively estimate body condition in many species, including cetaceans. Here, two measures of body condition were investigated in the harbor porpoise (Phocoena phocoena). First, the most informative morphometric body condition index was identified. The mass/length² ratio was the most appropriate morphometric index of 10 indices tested, explaining 50% of the variation in condition in stranded, male porpoises with different causes of death and across age classes (n = 291). Mass/length² was then used to evaluate a second measure, blubber cortisol concentration, as a metabolic condition marker. Cortisol is the main glucocorticoid hormone involved in the regulation of lipolysis and overall energy balance in mammals, and concentrations could provide information on physiological state. Blubber cortisol concentrations did not significantly vary around the girth (n = 20), but there was significant vertical stratification through the blubber depth with highest concentrations in the innermost layer. Concentrations in the dorsal, outermost layer were representative of concentrations through the full blubber depth, showed variation by sex and age class, and were negatively correlated with mass/length². Using this species as a model for live cetaceans from which standard morphometric measurements cannot be taken, but from which blubber biopsy samples are routinely collected, cortisol concentrations in the dorsal, outermost blubber layer could potentially be used as a biomarker of condition in free‐ranging animals.     

The Role of Canine Distemper Virus and Persistent Organic Pollutants in Mortality Patterns of Caspian Seals (Pusa caspica)

Persistent organic pollutants are a concern for species occupying high trophic levels since they can cause immunosuppression and impair reproduction. Mass mortalities due to canine distemper virus (CDV) occurred in Caspian seals (Pusa caspica), in spring of 1997, 2000 and 2001, but the potential role of organochlorine exposure in these epizootics remains undetermined. Here we integrate Caspian seal mortality data spanning 1971–2008, with data on age, body condition, pathology and blubber organochlorine concentration for carcases stranded between 1997 and 2002. We test the hypothesis that summed PCB and DDT concentrations contributed to CDV associated mortality during epizootics. We show that age is the primary factor explaining variation in blubber organochlorine concentrations, and that organochlorine burden, age, sex, and body condition do not account for CDV infection status (positive/negative) of animals dying in epizootics. Most animals (57%, n = 67) had PCB concentrations below proposed thresholds for toxic effects in marine mammals (17 µg/g lipid weight), and only 3 of 67 animals had predicted TEQ values exceeding levels seen to be associated with immune suppression in harbour seals (200 pg/g lipid weight). Mean organonchlorine levels were higher in CDV-negative animals indicating that organochlorines did not contribute significantly to CDV mortality in epizootics. Mortality monitoring in Azerbaijan 1971–2008 revealed bi-annual stranding peaks in late spring, following the annual moult and during autumn migrations northwards. Mortality peaks comparable to epizootic years were also recorded in the 1970s–1980s, consistent with previous undocumented CDV outbreaks. Gompertz growth curves show that Caspian seals achieve an asymptotic standard body length of 126–129 cm (n = 111). Males may continue to grow slowly throughout life. Mortality during epizootics may exceed the potential biological removal level (PBR) for the population, but the low frequency of epizootics suggest they are of secondary importance compared to anthropogenic sources of mortality such as fishing by-catch.     

Effects of Age,Adipose Percent,and Reproduction on PCB Concentrations and Profiles in an Extreme Fasting North Pacific Marine Mammal

Persistent organic pollutants, including polychlorinated biphenyls (PCBs), are widely distributed and detectable far from anthropogenic sources. Northern elephant seals (Mirounga angustirostris) biannually travel thousands of kilometers to forage in coastal and open-ocean regions of the northeast Pacific Ocean and then return to land where they fast while breeding and molting. Our study examined potential effects of age, adipose percent, and the difference between the breeding and molting fasts on PCB concentrations and congener profiles in blubber and serum of northern elephant seal females. Between 2005 and 2007, we sampled blubber and blood from 58 seals before and after a foraging trip, which were then analyzed for PCBs. Age did not significantly affect total PCB concentrations; however, the proportion of PCB congeners with different numbers of chlorine atoms was significantly affected by age, especially in the outer blubber. Younger adult females had a significantly greater proportion of low-chlorinated PCBs (tri-, tetra-, and penta-CBs) than older females, with the opposite trend observed for hepta-CBs, indicating that an age-associated process such as parity (birth) may significantly affect congener profiles. The percent of adipose tissue had a significant relationship with inner blubber PCB concentrations, with the highest mean concentrations observed at the end of the molting fast. These results highlight the importance of sampling across the entire blubber layer when assessing contaminant levels in phocid seals and taking into account the adipose stores and reproductive status of an animal when conducting contaminant research.     

DETERMINING PREGNANCY FROM BLUBBER IN THREE SPECIES OF DELPHINIDS

We quantified progesterone in 110 blubber samples from dolphins of known reproductive status in order to test the accuracy of a method to determine pregnancy status in wild cetaceans. The samples were collected from fishery‐bycaught delphinids of three species (Delphinus delphis, Lissodelphis borealis, and Lagenorhynchus obliquidens). We ascertained that blubber progesterone concentrations could clearly distinguish pregnant D. delphis (range 132–415 ng/g, mean 261 ng/g) from non‐pregnant mature and immature ones (range 0.92–48.2 ng/g, mean 15.2 ng/g). We found similar dramatic differences in L. borealis and L. obliquidens. These results were insensitive to various blubber sampling depths and anatomical sampling locations on the body, suggesting relative homogeneity of progesterone levels throughout the blubber. However, no trend was found in blubber progesterone concentration with fetal length, indicating that although blubber progesterone appears to distinguish pregnancy status, it is unlikely to differentiate pregnancy stage. Based on the findings presented here we suggest that this method, when coupled with projectile biopsy procedures, can be used to assess the pregnancy status of free‐ranging cetaceans and thus provide a new tool to determine pregnancy rates of wild populations.     

Blubber development in bottlenose dolphins (Tursiops truncatus)

Blubber, the lipid-rich hypodermis of cetaceans, functions in thermoregulation, buoyancy control, streamlining, metabolic energy storage, and locomotion. This study investigated the development of this specialized hypodermis in bottlenose dolphins (Tursiops truncatus) across an ontogenetic series, including fetuses, neonates, juveniles, subadults, and adults. Blubber samples were collected at the level of the mid-thorax, from robust specimens (n = 25) that stranded along the coasts of North Carolina and Virginia. Blubber was dissected from the carcass and its mass, and the depth and lipid content at the sample site, were measured. Samples were prepared using standard histological methods, viewed by light microscopy, and digital images of blubber captured. Images were analyzed through the depth of the blubber for morphological and structural features including adipocyte size, shape, and numbers, and extracellular, structural fiber densities. From fetus to adult, blubber mass and depth increased proportionally with body mass and length. Blubber lipid content increased dramatically with increasing fetal length. Adult and juvenile blubber had significantly higher blubber lipid content than fetuses, and this increase was reflected in mean adipocyte size, which increased significantly across all robust life history categories. In juvenile, subadult, and adult dolphins, this increase in cell size was not uniform across the depth of the blubber, with the largest increases observed in the middle and deep blubber regions. Through-depth counts of adipocytes were similar in all life history categories. These results suggest that blubber depth is increased during postnatal growth by increasing cell size rather than cell number. In emaciated adults (n = 2), lipid mobilization, as evidenced by a decrease in adipocyte size, was localized to the middle and deep blubber region. Thus, in terms of both lipid accumulation and depletion, the middle and deep blubber appear to be the most metabolically dynamic. The superficial blubber likely serves a structural role important in streamlining the animal. This study demonstrates that blubber is not a homogeneous tissue through its depth, and that it displays life history-dependent changes in its morphology and lipid content.     

Hepatic stellate cell lipid droplets: A specialized lipid droplet for retinoid storage

The majority of retinoid (vitamin A and its metabolites) present in the body of a healthy vertebrate is contained within lipid droplets present in the cytoplasm of hepatic stellate cells (HSCs). Two types of lipid droplets have been identified through histological analysis of HSCs within the liver: smaller droplets bounded by a unit membrane and larger membrane-free droplets. Dietary retinoid intake but not triglyceride intake markedly influences the number and size of HSC lipid droplets. The lipids present in rat HSC lipid droplets include retinyl ester, triglyceride, cholesteryl ester, cholesterol, phospholipids and free fatty acids. Retinyl ester and triglyceride are present at similar concentrations, and together these two classes of lipid account for approximately three-quarters of the total lipid in HSC lipid droplets. Both adipocyte-differentiation related protein and TIP47 have been identified by immunohistochemical analysis to be present in HSC lipid droplets. Lecithin:retinol acyltransferase (LRAT), an enzyme responsible for all retinyl ester synthesis within the liver, is required for HSC lipid droplet formation, since Lrat-deficient mice completely lack HSC lipid droplets. When HSCs become activated in response to hepatic injury, the lipid droplets and their retinoid contents are rapidly lost. Although loss of HSC lipid droplets is a hallmark of developing liver disease, it is not known whether this contributes to disease development or occurs simply as a consequence of disease progression. Collectively, the available information suggests that HSC lipid droplets are specialized organelles for hepatic retinoid storage and that loss of HSC lipid droplets may contribute to the development of hepatic disease.     

Variation in the fatty acid composition of blubber in Cape fur seals (<Emphasis Type="Italic">Arctocephalus pusillus pusillus</Emphasis>) and the implications for dietary interpretation

Analysis of the fatty acid (FA) composition of blubber is a valuable tool in interpreting the diet of marine mammals. This technique is based on the principle that particular FA present in prey can be incorporated largely untransformed into predator adipose tissue stores, thereby providing biochemical signatures with which to identify prey species. Several studies of phocid seals and cetaceans have documented vertical stratification in the FA composition of blubber such that inferences about diet may vary greatly depending on the layer of the blubber that is analysed. It is not known whether blubber in otariid seals (fur seals and sea lions) also displays vertical stratification in FA composition. Furthermore, it is not known whether the FA composition of blubber is uniform in these species. In the present study, the vertical and regional variation in FA composition of blubber was investigated in seven adult female Cape fur seals (Arctocephalus pusillus pusillus). The proportion of monounsaturated fatty acids (MUFA) was greater in the outer (43.6±1.3%) than inner portion (40.9±1.2%; t₂₀=5.59, P<0.001) whereas the proportions were greater in the inner than outer portions for saturated fatty acids (23.6±0.5% and 21.9±0.6%, respectively, t₂₀ = 5.31, P<0.001) and polyunsaturated fatty acids (PUFA, 35.5±0.7% and 34.5±0.7%, respectively, t₂₀ = 3.81, P < 0.001). There was an inverse relationship between MUFA and PUFA in the blubber, independent of sampling location. In addition, with the exception of the inner portion from non-lactating females, blubber from the mammary area had the highest proportions of 18:19c and total MUFA, followed by blubber from the rump and neck, suggesting that the deposition and mobilisation of blubber lipids may not be uniform around the body in otariid seals. These results support the need for blubber tissue to be sampled from the same site on animals, and to the full depth of the blubber layer, to minimise variation in FA profiles that could occur if different sites and depths were sampled. Such standardisation of sampling will further aid in interpreting diet in otariid seals using the FA Signature Analysis approach.     

How long is a piece of Strix? Methods in measuring and measuring the measurers

An experiment to quantify intra- and interobserver error in anatomical measurements found that interobserver measurements can vary by over 14% of mean specimen length; disparity in measurement increases logarithmically with the number of contributors; instructions did not reduce variation or measurement disparity; scale of the specimen influenced the precision of measurement (relative error increasing with specimen size); different methods of taking a measurement yielded different results, although they did not differ in terms of precision, and topographical complexity of the elements being considered may potentially influence error (error increasing with complexity). These results highlight concerns about introduction of noise and potential bias that should be taken into account when compiling composite datasets and meta-analyses.     

Relation between serum lipoperoxide concentrations and iron or copper status over one year in Cuban adult men

The aims of this study were to determine the relations between iron and copper status and lipid peroxidation at different periods over one year in low-income and low-energy intake healthy subjects. The study was conducted in 199 middle-aged healthy Cuban men from March 1995 to February 1996. Iron status was assessed by the determination of serum ferritin, transferrin saturation, whole blood hemoglobin and iron intakes. Copper status was evaluated by the determination of serum copper and copper intakes. Serum thiobarbituric acid substances (TBARS) determination was used as an index of lipid peroxidation. Rank correlations were observed between serum TBARS concentrations and iron or copper status indices at different periods. In period 3 (end of the rainy season), serum TBARS and ferritin concentrations were maximum whereas blood hemoglobin levels and iron intake were minimum. Serum TBARS concentrations were significantly higher than the reference values of the laboratory whereas, iron and copper status were within the reference ranges. These results suggested that iron and copper status may be associated with lipid peroxidation in subjects without metal overloads and that variations over the year needed to be taken in account.     

Vitamin A and E tissue distribution with comparisons to organochlorine concentrations in the serum, blubber and liver of the bowhead whale (Balaena mysticetus)

Vitamin A and E concentrations were determined in liver (n = 51), blubber (n = 23) and serum (n = 53) of subsistence-hunted bowhead whales (Balaena mysticetus), between 1998 and 2001. Retinol and alpha-tocopherol were the major forms of vitamins A and E detected, respectively. Liver contained the highest mean concentrations of vitamin A, followed by epidermis, blubber, and serum. Liver also contained the highest mean concentration of vitamin E, followed by serum, epidermis, and blubber. Stratification of retinol and tocopherol was examined throughout the blubber cores collected. Retinol concentrations were significantly higher in the epidermis than in the deeper blubber layers. Tocopherol concentrations were similar for epidermis and the intermediate layer of blubber. Both the epidermis and the intermediate layer of blubber had significantly higher tocopherol concentrations than the innermost and outermost blubber layers. Vitamin A and E concentrations were investigated with respect to gender and reproductive status of females (males, non-pregnant females, pregnant females), age groups and season of harvest. Certain persistent organic contaminants are known to have a negative effect on retinol concentration in serum of pinnipeds and cetaceans. Bowhead whales have relatively low concentrations of organochlorines (OCs) in comparison to other mysticete species. The relationships between serum, liver and blubber retinol and serum and blubber OC concentrations were examined with no significant correlations noted.     

Body regional distribution and stratification of fatty acids in the blubber of New Zealand sea lions: implications for diet predictions

Fatty acids (FAs) from blubber are often analysed to assess the diet of marine mammals. However, distribution of blubber FAs is not necessarily uniform along the body. It is therefore important to understand the deposition of dietary fat to be able to estimate the diet. We analysed the FA compositions of the thoracic ventral (T region) blubber of 28 New Zealand (NZ) sea lions Phocarctos hookeri by-caught by the southern arrow squid Nototodarus sloani fishery. Each blubber sample was divided into an inner and an outer layer. For 16 of these 28 animals, the pelvic dorsal (P) region was also sampled. The influence of body region and layer was statistically tested on the distribution of blubber FAs. We found minimal differences between the P and T regions (3 out of 29 FAs). The outer blubber layer was more concentrated in short-chain monounsaturated FAs, and less concentrated in saturated FAs, but the degree of stratification was small. Diet predictions from quantitative FA signature analysis (QFASA) applied on different body regions were similar. When applied to different blubber layers, QFASA gave some variation in the contribution of rattails (~25 % in outer blubber vs. ~12 % in inner blubber). Nonetheless, diet predicted from both layers was dominated by similar prey species: octopus, hoki and rattails. Hoki and rattails shared a similar ecological niche. Therefore, feeding ecology of NZ sea lions inferred from the inner or the outer blubber would lead to the same conclusions. In the case of NZ sea lions, the outer layer of blubber, if the only sample accessible, could be a useful tissue for diet inference from FAs.     

Pregnancy rate and biomarker validations from the blubber of eastern North Pacific blue whales

Hormonal biomarkers are useful indicators of mammalian reproductive and metabolic states. The present study validated and applied the use of progesterone and cortisol blubber assays for studies of blue whales from the Gulf of California, Mexico. In a validation study for pregnancy detection, blubber progesterone concentrations were correlated with pregnancy status for four female blue whales: three resighted with a calf the year following sampling and the fourth stranded with a fetus. The progesterone concentrations were significantly higher than those measured in juvenile whales (n = 3). In the application study, blubber samples from blue whales (51 noncalf females, 2 female calves, 48 noncalf males, and 1 male calf) with known sighting histories were analyzed. Putative pregnant females had elevated progesterone concentrations. Cortisol concentrations did not differ between male and female blue whales, or among females in different reproductive classes. After correcting for uncertain ages, hence maturity status, the pregnancy rate of noncalf females was 33.4% (95% CI 32.2%–34.3%). Although interpretation of hormone biomarkers must consider all physiological states that may influence progesterone concentrations, these results demonstrate the utility of pairing hormone biomarkers with sighting histories to help assess environmental or anthropogenic impacts on reproduction in blue whales.     

Body fat and condition in sperm whales,Physeter macrocephalus,from southern Australian waters

Blubber thickness (n=102) and lipid content (n=37) were measured in sperm whales from three mass stranding events on the west and north-west coasts of Tasmania, Australia in February 1998. Blubber thickness was highly variable, ranging from 43.0 to 168.0 mm (mean 98.4+/-18.4 mm) while lipid fat content, also highly variable, ranged from 16.19 to 89.34% (mean 49.2+/-17.9%). Blubber thickness was significantly and positively related to total length, but a blubber thickness index based on the residuals of this relationship was not related to age, sex or reproductive condition. No relationship was found between blubber thickness index and blubber lipid content, indicating that blubber thickness may not provide a comprehensive indication of body fat condition in sperm whales when only measured at a single site. Blubber lipid content was not related to total length, age or sex. Blubber lipid content was stratified vertically throughout the blubber layer, suggesting that the inner blubber layer may be a more active site for lipid deposition and mobilisation, while the outer blubber layer may serve in a structural or thermoregulatory role. The social structure and foraging ecology of this species may serve to minimise the need to rely on stored energy reserves to meet reproductive energy requirements. In addition, the broader role of blubber for structural, buoyancy and insulative functions coupled with high individual variability may cause a lack of obvious relationships between these variables and body size, age, sex and reproductive state in this species.     

Increased blubber cortisol in ice-entrapped beluga whales (<Emphasis Type="Italic">Delphinapterus</Emphasis><Emphasis Type="Italic">leucas</Emphasis>)

Entrapments of whales in sea ice occur occasionally in the Arctic and often last several weeks or months, resulting in emaciation or death of whales. These events provide a unique opportunity for investigating the physiological response to a prolonged or chronic stress in an otherwise healthy population of marine mammals. By measuring cortisol in blubber, a peripheral tissue, we expect to see a reflection of long-term or chronic stress rather than short-term or acute stress. Adipose tissue should be less subject to rapid changes compared to blood cortisol, reflecting stressors experienced over a longer period of time, and should not be affected by potential stress associated with sampling. We measured blubber cortisol of 29 beluga whales (Delphinapterus leucas) entrapped in November 2006 in Husky Lakes basin and 26 whales from the same population (Eastern Beaufort Sea) during regular seasonal harvests in July of 2006 and 2007. Mean cortisol concentrations (±SEM) were seven times higher in blubber from entrapped whales (1.76 ± 0.32 ng/g wet weight) compared to whales from regular seasonal harvests (0.26 ± 0.042 ng/g wet weight) and appeared to increase with whale age. Our results provide a measure of blubber cortisol from a prolonged stress and demonstrate blubber cortisol as a useful indicator of longer-term exposure to stress in beluga whales.     

Organochlorine contaminant levels in Eskimo harvested bowhead whales of arctic Alaska

Organochlorine (OC) levels in liver and blubber of 20 bowhead whales (Balaena mysticetus) collected during the Eskimo subsistence harvest at Barrow (Alaska, USA) in 1992 and 1993 are presented. Liver sum DDT (lipid weight) was significantly greater in male whales than in females. Most of the organochlorines measured were at higher levels in longer (older) than in shorter (younger) males. For female bowhead whales, hexachlorobenzene and lipid levels decreased and other OC levels did not change significantly with increasing length. Most organochlorine contaminants have low concentrations in tissues of the bowhead whale compared to concentrations in tissues of other cetaceans, especially Odontocetes. Based on allowable daily intakes (ADI) levels established by the Canadian Northern Contaminants Program (Ottawa, Ontario, Canada) "safe" levels of blubber to consume were calculated. Chlordane levels in bowhead whale blubber results in the most restrictive consumption amount (50 g blubber/day). We expect no adverse effects related to these organochlorine contaminants to occur in bowhead whales or in consumers of their tissues. However, investigation of low level chronic exposure effects and a more rigorous assessment of histopathology, biomarkers, and immune status in the bowhead whale would be required to conclude "no effect" with more certainty.     

Blubber fatty acid composition of bowhead whales, Balaena mysticetus: Implications for diet assessment and ecosystem monitoring

Fatty acids (FA) have a diversity of structures that are transferred with little modification through food webs, making them valuable in assessing diets of animals that cannot be directly observed feeding. Before using FA to estimate diets, it is necessary to evaluate variation in FA signatures within and among individuals of a given species. To begin assessing diets and foraging of western Arctic bowhead whales (Balaena mysticetus), we examined the FA in blubber of 64 bowheads taken in the spring and fall subsistence hunts in 1997–2002 at Barrow and Kaktovik, Alaska. We found no significant differences in FA characteristics of inner blubber layers taken from either duplicate samples on the dorsal surface, or between dorsal and ventral sites. Significant differences were found in the FA composition between inner and outer layers of blubber at the same body site. We also found age, season and year to have significant effects on FA composition; however, gender was not found to be significant. While the importance of the Beaufort Sea as a feeding ground of bowhead whales remains uncertain, our results indicate that adults and sub-adults foraged to some extent on different prey and that both age classes consumed copepods there in summer at sufficient levels to significantly alter their blubber FA profiles. Both of these findings correspond with dietary conclusions reached from the analysis of stomach contents. Furthermore, we found compelling evidence that yearly variation in bowhead FA reflect changes in FA compositions of phytoplankton at the base of the food web, probably in response to climate variation. Variability in phytoplankton-derived FA in blubber was correlated significantly with yearly mean values of the Pacific Decadal Oscillation. FA in bowhead whale blubber, therefore, might be used to monitor effects of climate change on lower trophic levels and production processes in the western Arctic.     

Hormone,vitamin and contaminant status during the moulting/fasting period in ringed seals (Pusa [Phoca] hispida) from Svalbard

This study investigates the potential effects of moulting, and the concomitant period of fasting undertaken by ringed seals, on hormone, vitamin and contaminant status in adult animals in a population from Svalbard, Norway, which has relatively low contaminant levels. Concentrations of circulating total and free thyroxine and triiodothyronine, circulating and hepatic vitamin A, hepatic persistent organic pollutants and their circulating hydroxyl metabolites were higher in moulting seals compared to pre-moulting seals. The opposite trend was observed for body condition, circulating calcitriol levels and hepatic mRNA expression of thyroid hormone receptor β. No differences were observed for circulating or hepatic vitamin E levels or hepatic mRNA expressions for deioidinase 1 or 2, or retinoic acid receptor α between the two seal groups. The observed differences are likely the result of increased metabolic rates required during moulting to maintain thermal balance and replace the pelage, in combination with mobilization of lipid soluble compounds from blubber stores during the fasting period that is associated with moulting. The present study shows that contaminant levels and their relationships with physiological or endogenous variables can be highly confounded by moulting/fasting status. Thus, moulting status and body condition should be taken into consideration when using variables related to thyroid, calcium or vitamin A homeostasis as biomarkers for contaminant effects.     

Production of Nicotinamide Adenine Dinucleotide Glycohydrolases by Bacteria

The distribution of heavy metals (Fe, Mn, Zn, Cu, Pb, Ni and Cd) were investigated in various organs and tissues of striped dolphin, Stenella coeruleoalba. The animals were caught alive at Taiji, on the coast of Kii Peninsula, during the open season in December 1978. Determination of the metals was made by atomic absorption spectrophotometry and significant differences of metal concentration in the positions of the muscle, blubber and skin, respectively, were observed. The front ventral muscle of matured dolphins showed the highest concentrations of Zn and Cd and lowest Fe when compared to other parts of the muscle. Most of the metals recorded relatively low concentrations in melon rather than in the other lipid layers of blubber. In skin tissue, the concentrations of Fe, Mn and Zn were significantly higher in black-colored skin than in white skin. Moreover, a difference in the concentrations of metals according to bone position was observed. In general, high concentrations of most of the metals were found in liver, kidney and bone, with low concentrations in brain and the lipid layer of blubber. Furthermore, relatively high concentrations of Cu, Mn and Zn were found in skin, and for Mn, Zn, Ni and Cd it was likewise in pancreas and the reproductive organs. Based upon these results, the nature of the organ(s) of a dolphin that has to be selected for ecological and hygienic comparison was discussed.     

Why do species vary in their rate of molecular evolution?

Despite hopes that the processes of molecular evolution would be simple, clock-like and essentially universal, variation in the rate of molecular evolution is manifest at all levels of biological organization. Furthermore, it has become clear that rate variation has a systematic component: rate of molecular evolution can vary consistently with species body size, population dynamics, lifestyle and location. This suggests that the rate of molecular evolution should be considered part of life-history variation between species, which must be taken into account when interpreting DNA sequence differences between lineages. Uncovering the causes and correlates of rate variation may allow the development of new biologically motivated models of molecular evolution that may improve bioinformatic and phylogenetic analyses.     

DETERMINATION OF PREGNANCY STATUS FROM BLUBBER SAMPLES IN MINKE WHALES (BALAENOPTERA ACUTOROSTRATA)

Measurement of progesterone concentration in blubber was developed as a method to detect pregnancy in minke whales. Progesterone was extracted and quantified from blubber samples of minke whale carcasses by radioimmunoassay. Results showed a highly significant difference (almost 60-fold) between blubber progesterone concentrations of anatomically determined pregnant females versus non-pregnant female or male carcasses. The results of the study suggest that the blubber progesterone concentrations might be used to determine pregnancy status in free-ranging whales.