Taxonomic review of Scinax fuscomarginatus (Lutz, 1925) and related species (Anura; Hylidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Dietary media for mass rearing of rusty grain beetle, Cryptolestes ferrugineus (Stephens) and flat grain beetle, Cryptolestes pusillus (Schonherr) (Coleoptera: Cucujidae)

We developed a suitable diet for mass rearing of Cryptolestes ferrugineus (Stephens) populations under laboratory conditions. Recently, this pest has developed strong level of resistance to phosphine in Australia, and therefore, a significant amount of research has been directed towards its management. In total, nineteen grain-based diets, containing rolled oats, various combinations of cracked grains and flours of wheat, sorghum, maize and barley were tested. Each diet contained a small proportion of wheat germ (4.5% w/w) and torula yeast (0.5% w/w). Experiments were conducted at fixed temperature and relative humidity regimes of 30 ± 2 °C and 70 ± 2%, respectively, and replicated three times. Adults (n = 40) of a laboratory strain of C. ferrugineus were introduced into each diet, removed after 14 days and total numbers of live adult progeny were recorded. The following diets resulted in highest live progeny production: barley flour (95%) (607.67 ± 11.21) = rolled oats (75%) + cracked sorghum (20%) (597.33 ± 33.79) ≥ wheat flour (47.5%) + barley flour (47.5%) (496.67 ± 52.93) > cracked sorghum (95%) (384.00 ± 60.66). The performance of these four diets was then tested with field-collected populations of C. ferrugineus and Cryptolestes pusillus (Schonherr). The diets based on rolled oats + cracked sorghum, wheat flour + barley flour, and barley flour alone consistently produced highest progeny numbers in field-collected populations of both species, with mean progeny numbers ranging from 359.9 to 478.5. The multiplication of C. pusillus was significantly higher than C. ferrugineus on all four diets. Our findings will help in mass rearing of healthy cultures of C. ferrugineus and C. pusillus that will greatly facilitate laboratory and field research and in particular, in developing management tactics for these species.

Trawling Operations and South African (Cape) Fur Seals, Arctocephalus pusillus pusillus

South African (Cape) fur seals, Arctocephalus pusillus pusillus, interact with the South African trawl fisheries-offshore demersal, inshore demersal, and midwater fisheries. These interactions take thef ollowing forms: Seals take or damage netted fish, on particular vessels they become caught in the propeller, seals drown in the nets, live seals come aboard and may be killed. Except in specific cases of seals damaging particular trawler propellers, interactions result in little cost to the offshore and midwater trawl fisheries. For the inshore fishery, seals damage fish in the net at an estimated cost in excess of R69, 728 (US$18,827) per year, but this is negligible (0.3%) in terms ofthe value of the fishery. Seal mortality is mainly caused by drowning in trawl nets and ranges from 2,524 to 3,636 seals of both sexes per year. Between 312 and 567 seals are deliberately killed annually, but this most likely takes place only when caught and they enter the area below deck, where they are difficult to remove, and pose a potential threat to crew safety. Overall, seal mortality during trawling operations is negligible (0.4-0.6%) in terms of the feeding population of seals in South Africa.

Biology, population dynamics, management and conservation of deep water lantern sharks, Etmopterus spinax and Etmopterus pusillus (Chondrichthyes: Etmopteridae) in southern Portugal (northeast Atlantic)

Tese dout., Ciências e Tecnologias das Pescas, Universidade do Algarve, 2007

Biologia alimentar do Etmopterus pusillus no Algarve

Dissertação mest., Biologia Marinha, Universidade do Algarve, 2007

Energy substrate utilization during nightly vocal activity in three species of Scinax (Anura/Hylidae)

Anuran amphibians exhibit different patterns of energy substrate utilization that correlate with the intensity of vocal and locomotor activities. Given the remarkable differences among species in breeding and feeding strategies, and the different ways energy is used in the whole animal, the suggested correlations between calling and locomotor behavior and the level of energy substrates in the muscles responsible for such activities are more complex than previously reported. We explored the relationships between calling and locomotor behavior and energy supply to trunk and hindlimb muscles, respectively, within the ecologically diverse tree-frog genus Scinax. Specifically, we measured the relative amount of carbohydrates and lipids in these two groups of muscles, and in the liver of three species of Scinax that differ in vocal and locomotor performance, and compared our results with those of two other species for which comparable data are available. We also compared the contents of lipids and carbohydrates of conspecific males collected at the beginning and after 4 h of calling activity. The stomach content to potential feeding opportunities across species was also assessed in both groups of males. Scinax hiemalis and S. rizibilis exhibit comparatively low and episodic calling during long periods of activity whereas S. crospedospilus calls at higher rates over shorter periods. Male S. hiemalis had highest levels of trunk muscle glycogen followed by those of S. rizilbilis and S. crospedospilus, respectively. There was no correlation between total lipid content in trunk muscle and calling rate among different species, suggesting that other metabolic aspects may be responsible for the energetic support for vocal activity. The levels of lipids and carbohydrates in trunk and hindlimb muscles and liver of males collected at the beginning and 4 h into the calling period were similar across species, so the extent of energetic reserves does not appear to constrain vocal or locomotor activity. Finally, we found exceptionally high levels of carbohydrates and lipids in the liver of S. rizibilis, a trait perhaps related to a long and demanding breeding period.

Variation in the fatty acid composition of the blubber in Cape fur seals (Arctocephalus pusillus pusillus) 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 18:1ω9c 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.

The physiological and behavioural development of diving in Australian fur seal (Arctocephalus pusillus doriferus) pups

The physiological and behavioural development of diving was examined in Australian fur seal (Arctocephalus pusillus doriferus) pups to assess whether animals at weaning are capable of exploiting the same resources as adult females. Haematocrit, haemoglobin and myoglobin contents all increased throughout pup development though total body oxygen stores reached only 71% of adult female levels just prior to weaning. Oxygen storage components, however, did not develop at the same pace. Whereas blood oxygen stores had reached adult female levels by 9 months of age, muscle oxygen stores were slower to develop, reaching only 23% of adult levels by this age. Increases in diving behaviour corresponded to the physiological changes observed. Pups spent little time (<8%) in the water prior to moulting (age 1–2 months) whereas following the moult, they spent >27% of time in the water and made mid-water dives (maximum depth 35.7 ± 2.9 m) with durations of 0.35 ± 0.03 min. By 9 months (just prior to weaning), 30.5 ± 9.3% of all dives performed were U-shaped benthic dives (maximum depth 65.0 ± 6.0 m) with mean durations of 0.87 ± 0.25 min, significantly shorter than those of adult females. These results suggest that while Australian fur seal pups approaching the age of weaning are able to reach similar depths as adult females, they do not have the physiological capacity to remain at these depths for sufficient durations to exploit them to the same efficiency.

Species-specific characteristics and individual variation of the bark call produced by male Australian fur seals (Arctocephalus pusillus doriferus)

Australian fur seals Arctocephalus pusillus doriferus are colonial breeding animals forming dense social groups during the breeding season. During this time, males establish and defend territories through physical conflicts, stereotyped posturing and vocalisations. While vocalisations are suggested to play an important role in male recognition systems, it has received little attention. Recordings of nine adult male Australian fur seals were made during the 2000 and 2001 breeding seasons at Kanowna Island (39° 10’S, 146° 18' E), Bass Strait, Australia. The in-air bark vocalisations of territory-holding males were used to characterise the Bark Call and to determine whether males produce individually distinct calls, which could be used as a basis for vocal recognition. Seventeen frequency and temporal variables were measured from a total of 162 barks from nine individual males. The Bark Series was more reliably classified (83%) to the correct caller compared to the Bark Unit. This was assigned with less certainty (68%), although the classification was still relatively high. Findings from this study indicate that there is sufficient stereotypy within individual calls, and sufficient variation between them, to enable vocal recognition in male Australian fur seals.

Pup production and population trends of the Australian fur seal (Arctocephalus pusillus doriferus)

We estimated the number of live Australian fur seal pups using capture-markresights, direct ground counts, or aerial photography at all breeding sites following the pupping season of November-December 2002. Pups were recorded at 17 locations; nine previously known colony sites, one newly recognized colony and seven haul-out sites where pups are occasionally born. In order of size, the colonies were Lady Julia Percy Island (5,899 pups), Seal Rocks (4,882), The Skerries (2,486), Judgment Rocks (2,427), Kanowna Island (2,301), Moriarty Rocks (1,007), Reid Rocks (384), West Moncoeur Island (257), and Tenth Island (124). The newly recognized site was Rag Island, in the Cliffy Group, where we recorded 30 pups. We also recorded pups at the following haul-out sites: Cape Bridge-water (7 pups), Bull Rock (7), Wright Rock (5), Twin Islet (1), The Friars (1), He des Phoques (1), and Montague Island (1). In total, we estimate there were 19,819 (SE = 163) live pups at the time of the counts. We discuss trends in pup numbers and derive current population estimates for the Australian fur seal.

Species-specific cell-matrix interactions are essential for differentiation of alveoli like structures and milk gene expression in primary mammary cells of the Cape fur seal (Arctocephalus pusillus pusillus)

Few models are in place for analysis of extreme lactation patterns such as that of the fur seals which are capable of extended down regulation of milk production in the absence of involution. During a 10–12 month lactation period, female fur seals suckle pups on shore for 2–3 days, and then undertake long foraging trips at sea for up to 28 days, resulting in the longest intersuckling bouts recorded. During this time the mammary gland down regulates milk production. We have induced Cape fur seal (Arctocephalus pusillus pusillus) mammary cells in vitro to form mammospheres up to 900 μm in diameter, larger than any of their mammalian counterparts. Mammosphere lumens were shown to form via apoptosis and cells comprising the cellular boundary stained vimentin positive. The Cape fur seal GAPDH gene was cloned and used in RT-PCR as a normalization tool to examine comparative expression of milk protein genes (αS2-casein, β-lactoglobulin and lysozyme C) which were prolactin responsive. Cape fur seal mammary cells were found to be unique; they did not require Matrigel for rapid mammosphere formation and instead deposited their own matrix within 2 days of culture. When grown on Matrigel, cells exhibited branching/stellate morphogenesis highlighting the species-specific nature of cell–matrix interactions during morphological differentiation. Matrix produced in vitro by cells did not support formation of human breast cancer cell line, PMC42 mammospheres. This novel model system will help define the molecular pathways controlling the regulation of milk protein expression and species specific requirements of the extracellular matrix in the cape fur seal.

Growth and condition in Australian fur seals Arctocephalus pusillus doriferus (Carnivora : Pinnipedia)

Mass and length growth models were determined for male (n = 69) and female (n = 163) Australian fur seals (Arctocephalus pusillus doriferus) collected at a breeding colony on Seal Rocks (38˚31′S, 145˚06′E), Bass Strait, in south-east Australia, between February and November during 1970–72. Growth was best described by the logistic model in males and the von Bertalanffy model in females. Asymptotic mass and length were 229 kg and 221 cm for males, and 85 kg and 163 cm for females. In all, 95% of asymptotic mass and length were attained by 11 years and 11 years, respectively, in males compared with 9 years and 5 years, respectively, in females. Males grew in length faster than females and experienced a growth spurt in mass coinciding with the onset of puberty (4–5 years). The onset of puberty in females occurs when approximately 86% of asymptotic length is attained. The rate of growth and sexual development in Australian fur seals is similar to (if not faster than) that in the conspecific Cape fur seal (A. p. pusillus), which inhabits the nutrient-rich Benguela current. This suggests that the low marine productivity of Bass Strait may not be cause of the slow rate of recovery of the Australian fur seal population following the severe over-exploitation of the commercial sealing era. Sternal blubber depth was positively correlated in adult animals with a body condition index derived from the residuals of the mass–length relationship (males: r² = 0.38, n = 19, P < 0.001; females: r² = 0.22, n = 92, P < 0.001), confirming the validity of using such indices on otariids. Sternal blubber depth varied significantly with season in adult animals. In males it was lowest in winter and increased during spring prior to the breeding season (r² = 0.39, n = 19, P < 0.03) whereas in females it was greatest during winter (r² = 0.05, n = 122, P< 0.05).

Dive behaviour, foraging locations and maternal-attendance patterns in Australian fur seals (Arctocephalus pusillus doriferus)

The dive behaviour, foraging locations, and colony-attendance patterns of female Australian fur seals (Arctocephalus pusillus doriferus) from Kanowna Island (39°10'S, 146°18'E) in Bass Strait, southeastern Australia, were determined throughout lactation during 1997–1999. Foraging-trip durations increased as lactation progressed, being shortest in summer (3.71 ± 0.24 days; mean ± 1 SE) and longest in winter (6.77 ± 0.57 days, P < 0.05), but maternal-attendance periods did not differ in duration (1.70 ± 0.10 days, P > 0.5). Individual mean attendance periods and trip durations were positively correlated (r² = 0.21, P < 0.005). Diving commenced shortly after seals left the colony (2.6 ± 0.4 h), was continuous for long periods (up to 36 h), occurred mostly during daylight hours, and lacked regular diel variation in depth. The majority of dives (78%) were typically U-shaped and reached depths corresponding to the prevailing depths in Bass Strait (65–85 m), indicating that these animals forage mostly on the benthos of the shallow continental shelf in this region. Such behaviour is unusual for fur seals but is reminiscent of that of some sea lion species. Mean dive durations varied between 2.0 and 3.7 min (maximum 8.9 min) and the theoretical aerobic dive limit (3.91–4.26 min) was exceeded on 17.3% of dives. Dive frequency (8.3 ± 0.6/h) and the proportion of time at sea spent diving (40.7 ± 2.1%) were weakly negatively related to the duration of the foraging trip (r²= 0.07, P < 0.004, and r² = 0.13, P < 0.0001, respectively). Data from at-sea locations showed that lactating females forage almost exclusively within Bass Strait during all seasons.

Chemical characterisation of the oligosaccharides in hooded seal (Cystophora cristata) and Australian fur seal (Arctocephalus pusillus doriferus) milk

Carbohydrates were extracted from hooded seal milk, Crystophora cristata (family Phocidae). Free oligosaccharides were separated by gel filtration and then purified by ion exchange chromatography, gel filtration and preparative thin layer or paper chromatography and their structures determined by ¹H-NMR. The hooded seal milk was found to contain inositol and at least nine oligosaccharides, most of which had lacto-N-neotetraose or lacto-N-neohexaose as core units, similar to those in milk of other species of Carnivora such as bears (Ursidae). Their structures were as follows: Gal(β1-4)Glc (lactose); Fuc(α1-2)Gal(β1-4)Glc (2′-fucosyllactose); Gal(β1-4)GlcNAc(β1-3)Gal(β1-4)Glc (lacto-N-neotetraose); Fuc(α1-2)Gal(β1-4)GlcNAc(β1-3)Gal(β1-4)Glc (lacto-N-fucopentaose IV); Gal(β1-4)GlcNAc(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(1-4)Glc (lacto-N-neohexaose); Fuc(α1-2)Gal(β1-4)GlcNAc(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (monofucosyl lacto-N-neohexaose a); Gal(β1-4)GlcNAc(β1-3)[Fuc(α1-2)Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (monofucosyl lacto-N-neohexaose b); Fuc(α1-2)Gal(β1-4)GlcNAc(β1-3)[Fuc(α1-2)Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (difucosyl lacto-N-neohexaose); Gal(β1-4)GlcNAc(β1-3)Gal(β1-4)GlcNAc(β1-3)Gal(β1-4)Glc (para lacto-N-neohexaose); Fuc(α1-2)Gal(β1-4)GlcNAc(β1-3)Gal(β1-4)GlcNAc(β1-3)Gal(β1-4)Glc (monofucosyl para lacto-N-neohexaose). Milk of the Australian fur seal, Arctophalus pusillus doriferus (family Otariidae) contained inositol but no lactose or free oligosaccharides. These results, therefore, support the hypothesis that the milk of otariids, unlike that of phocids, contains no free reducing saccharides.