Reproductive aspects of the Atlantic angel shark Squatina dumeril in the southern Caribbean Sea.

The maturity and reproduction of the Atlantic angel shark Squatina dumeril were assessed using 77 females (29·2-110·4 cm total length; LT ) and 269 males (58·7-108·2 cm LT ) harvested by artisanal gillnetters off Venezuela. The biased sex ratio implied segregation or sex-specific gear selectivity. Based on the development of the reproductive tract, 50% LT at sexual maturity (LT50 , mean ± s.e.) for females and males were estimated at 86·14 ± 0·64 and 81·55 ± 0·12 cm, respectively. Uterine fecundity ranged between one and six and with a maximum embryo size of 25·7 cm LT . Gravid females were observed from August to December, including those close to parturition and while the gestation period was not confirmed, the size of ovarian follicles among some specimens implied protraction. The low fecundity of the species supports close monitoring of catches.

Interactions between Manta birostris and Sotalia guianensis in a World Heritage listed Brazilian estuary.

During 1442 h of visual observations over 7 years throughout the World Heritage listed Paranaguá estuarine complex, Brazil, seven occurrences of interactions were observed at a single location involving breaching Manta birostris displacing schools of teleosts, which were subsequently preyed upon by Sotalia guianensis. Although the interactions were not definitively categorized as being amensal, commensal or mutual, their restriction to isolated space (adjacent to a protected area) and time (summer) supports previous assertions the area is important to regional productivity and the continuation of protected-area status.

Sublethal effects of angling and release on golden perch Macquaria ambigua: implications for reproduction and fish health.

The present study tested the hypothesis of no delayed sublethal effects of mild angling and release on the feeding, growth, somatic condition and gonadal development of golden perch Macquaria ambigua during gametogenesis. Subsamples of adult M. ambigua (n = 17-21 of 207), originally captured from the wild and stocked into ten 0·1 ha earthen ponds, were angled and released during early and late gametogenesis. Wild samples that were concurrently collected throughout the experiment underwent rapid and synchronous gonadal development and many spawned. While no spawning occurred in the ponds, most M. ambigua underwent normal gonadal development to maturity, including the angled fish. Angled fish also fed, maintained condition and actually grew faster than non-angled captive controls. Although females that were angled during late gametogenesis more readily ingested and retained baited hooks, neither their subsequent condition nor gonadal development was significantly affected. The predominance of null results was attributed to the combined effects of the flexible reproductive strategy of M. ambigua, the benignness of mouth hooking and immediate release, and possible methodological issues arising from differential hooking success of more aggressive and resilient individuals. The findings support earlier catch-and-release research, but contrast with reports of acute reproductive effects following capture and handling for aquaculture broodstock. This discrepancy highlights the need for research to specifically address welfare questions relevant to recreational fisheries across various species and angling scenarios.

Seasonal and environmental influences on recruitment patterns and habitat usage among resident and transient fishes in a World Heritage Site subtropical estuary.

This study investigated whether the fish communities inhabiting shallow non-vegetated habitats in two divergent bays in a subtropical World Heritage Site estuarine system differed according to wet (spring-summer) and dry (autumn-winter) seasons or polyhaline and mesohaline zones, within the broader objective of facilitating spatio-temporal management. Species richness (total of 74 taxa; total length, LT = 11-552 mm) and abundance (51 109 individuals) were mostly greater in the wet than dry season and in polyhaline than mesohaline areas. There was a major effect of rainfall on recruitment, particularly among transient fishes, which could be the result of enhanced survival of young via greater productivity (food resources) and protection from predators (via turbidity reducing visual cues). Salinity had strong interactive effects with rainfall and temperature in one bay, with greater species richness and overall abundances as well as large abundances of four key species [Anchoa januaria and Atherinella brasiliensis (pelagic residents), Cetengraulis edentulus (pelagic transient) and Diapterus rhombeus (demersal transient)] during the wet season in polyhaline areas; possibly reflecting a biodiversity hotspot that might be affected by distance to the estuary mouth and convergence hydrology. Regionally, the results support enforcing spatio-temporal restrictions to minimize anthropogenic activities within statutory (but not always enforced) protected areas. Globally, the data reiterate the need to identify and understand biotic and abiotic effects on estuarine ichthyofaunal distributions and abundances as a precursor to their management.

Mapping the spatio-temporal distribution of threatened batoids to improve conservation in a subtropical estuary.

The spatio-temporal distributions of four batoid species were examined in a subtropical estuary. Fluvial gradient was the most important factor explaining abundances, reflecting positive relationships with either salinity or distance from urbanised areas that were consistent across seasons and depths. The results support existing protected areas.

Clinical signs of barotrauma in golden perch, Macquaria ambigua (Richardson), and associated effects on post-release mortality and health.

This study assessed the effects of different retrieval depths (2, 10 or 20 m), surface intervals (none or 15 min) and release methods (untreated, vented or recompressed) on the incidence of external and internal clinical signs of barotrauma (ECSB and ICSB) and post-release mortality in golden perch, Macquaria ambigua (Richardson). Fish were assessed for ECSB before and after surface intervals and either monitored for mortality over 3 days in two deep cages or killed for internal examination. When all fish were left untreated, short-term mortality increased with retrieval depth from 0% and 4.2% among 2 and 10-m fish, respectively, to 19.2% among 20-m fish; while surface interval only affected the incidence of two ECSB (excess buoyancy and a prolapsed cloaca). Mortality was also greater among 20-m fish that were subjected to a 15-min surface interval and left untreated (22.2%) or vented (22.2%) than those that were recompressed (5.6%). Of the ECSB, only exophthalmia was associated with increased mortality, with half of the affected fish dying. However, many fish retrieved from 10 and 20 m also sustained numerous ICSB, including compressed gonads or vital organs and ruptured or collapsed, haemorrhaging swimbladders that remained deflated for up to 3 days after release.

Host-defence-related proteins in cows' milk.

Milk is a source of bioactive molecules with wide-ranging functions. Among these, the immune properties have been the best characterised. In recent years, it has become apparent that besides the immunoglobulins, milk also contains a range of minor immune-related proteins that collectively form a significant first line of defence against pathogens, acting both within the mammary gland itself as well as in the digestive tract of the suckling neonate. We have used proteomics technologies to characterise the repertoire of host-defence-related milk proteins in detail, revealing more than 100 distinct gene products in milk, of which at least 15 are known host-defence-related proteins. Those having intrinsic antimicrobial activity likely function as effector proteins of the local mucosal immune defence (e.g. defensins, cathelicidins and the calgranulins). Here, we focus on the activities and biological roles of the cathelicidins and mammary serum amyloid A. The function of the immune-related milk proteins that do not have intrinsic antimicrobial activity is also discussed, notably lipopolysaccharide-binding protein, RNase4, RNase5/angiogenin and cartilage-glycoprotein 39 kDa. Evidence is shown that at least some of these facilitate recognition of microbes, resulting in the activation of innate immune signalling pathways in cells associated with the mammary and/or gut mucosal surface. Finally, the contribution of the bacteria in milk to its functionality is discussed. These investigations are elucidating how an effective first line of defence is achieved in the bovine mammary gland and how milk contributes to optimal digestive function in the suckling calf. This study will contribute to a better understanding of the health benefits of milk, as well as to the development of high-value ingredients from milk.

Resilience of inshore, juvenile snapper Pagrus auratus to angling and release.

This study assessed the mortality of 157 snapper Pagrus auratus (9-29 cm, total length, L(T) ) after being conventionally angled and then released into cages (along with 48 controls) for 4 days off south-eastern Australia. Fatalities were restricted to 12 angled fish (7·6%) and mostly attributed to the ingestion of hooks and especially their subsequent removal, which caused substantial blood loss and immediate death. Hook ingestion was significantly biased towards smaller fish (<21 cm L(T)) and attributed to a lower chance of anglers initially detecting these individuals on the line (allowing them to consume more of the baits). While mortalities might be reduced in future via (1) choosing terminal rigs that promote mouth hooking and (2) cutting the line on any-hook ingested fish, the results nevertheless validate releasing unwanted angled inshore juvenile P. auratus as a means for managing their exploitation.

Effects of barotrauma and mitigation methods on released Australian bass Macquaria novemaculeata.

The effects of barotrauma on the short-term mortality and physical condition of Australian bass Macquaria novemaculeata were investigated after being: (1) conventionally angled and released during two tournaments in deep impoundments and (2) released untreated or subjected to venting or recompression during a manipulative experiment. All fish were released into two 20 m deep bathy-cages and monitored for 3 days. Of 238 M. novemaculeata angled during the tournaments, 43 (18·1%) had clinical signs of barotrauma or were vented and five of these later died (11·6% mortality). Catch histories varied significantly between both barotrauma and non-barotrauma fish and tournaments, but only hook ingestion significantly influenced mortality (P < 0·05). During the manipulative experiment, venting significantly influenced mortality (13·3%) compared to no treatment or recompression (no deaths). Magnetic resonance images and dissections of barotrauma fish indicated large variation among clinical signs. On the basis of these results, wherever possible M. novemaculeata suffering barotrauma should be immediately released with no treatment. Fish that are unable to resubmerge should be recompressed, while those held in live wells and released in shallow water should be vented.

The abundance of milk cathelicidin proteins during bovine mastitis.

Current on-farm methods for detecting mastitis in dairy cows have limitations with their specificity and sensitivity, particularly at an early stage of infection. There is therefore a need to explore new approaches for detecting early and subclinical mastitis. This study examined the expression of a group of neutrophil-specific proteins, the cathelicidins, in milk samples from naturally occurring as well as experimentally induced mastitis infections. Immunoblot analysis indicated that cathelicidin proteins are only observed in infected quarters and demonstrate a high correlation with somatic cell count (SCC) during the onset of infection. In most of the infections examined, cathelicidin was detected prior to the observation of clinical symptoms and at SCC counts as low as 6.2 × 10(3)cells/mL. In naturally occurring mastitis the correlation between cathelicidin and infection status is not as strong, with 25% of pathogen-positive milk samples containing no detectable cathelicidin. This may reflect the varying levels of neutrophil concentration and activity at different stages or severities of infection. Our results indicate that milk cathelicidin levels increase following intramammary infection and cathelicidin-based biomarkers may assist in the detection of preclinical mastitis or determining the stage of infection.

Effects of salinity and anatomical hook location on the mortality and physiological response of angled-and-released sand whiting Sillago ciliata.

Three experiments were done with sand whiting Sillago ciliata: the first two assessed the short-term mortality and physiological response of individuals after being mouth hooked and then subjected to rapid changes in salinity, while the third experiment investigated their longer-term fate after ingesting hooks (independent of salinity changes). In experiment one, 48 tanks containing a single S. ciliata were randomly assigned as either one of three treatments or a control. The fish in treatments one and two were exposed to salinity changes during their angling and subsequent release while those in treatment three were only subjected to angling and air exposure. Control fish remained untouched. Fish were then monitored for up to 6 days for mortalities before blood samples were taken to determine concentrations of plasma cortisol and glucose. Blood samples were also taken from five wild-caught fish to provide baseline estimates of the above variables. None of the treatment or control fish died over the 6 days, and there were no significant differences in blood cortisol or glucose between treatment, control and wild fish. In experiment two, 102 S. ciliata and 52 experimental tanks were used. The treatments were repeated as above, however, six individuals from each treatment and control group were removed and sampled for blood (and then glucose and cortisol) at 0, 24, 48 and 72 h post release. Some changes in behaviour due to the salinity changes occurred as well as a significant main effect of time for cortisol, with all fish having significantly elevated acute stress at the first sample time. In experiment three, 52 S. ciliata were placed into individual tanks. Twenty-six of these fish were allowed to ingest baited J-hooks, played for 60 s, removed from their tanks, and then released after their lines were cut (50 mm from their mouths). Control fish were not touched. All fish were then monitored over 21 days. Six of the treatment fish died (between 3 h and 14 days), while the remaining hooked fish resumed feeding within 5 days and 25% ejected their hooks (between 1 and 19 days). It was concluded that (1) salinity and mouth hooking had few independent or interactive effects on the mortality or physiological response of angled-and-released S. ciliata and (2) while hook ingestion caused some mortalities, the protracted physiological effects were limited, with all surviving fish resuming feeding and some eventually ejecting their hooks.

Effects of angling on post-release mortality, gonadal development and somatic condition of Australian bass Macquaria novemaculeata.

This study assessed the protracted effects of two angling treatments (mild and harsh) on the post-release mortality, gonadal development and somatic condition of Australian bass Macquaria novemaculeata in a pond-based experiment. Angled fish were destructively sampled (along with controls) three times: immediately before, and 1 or 4 weeks after release into cages. Compared with the other groups, harshly angled fish had similar, low immediate and short-term mortalities (< or =5%), but significant delayed mortality (25%) and fed only minimally. None of these fish ejected ingested hooks. Forty-six per cent of captive fish (across all groups) and 20% of wild fish had non-ripening gonads (stage I) prior to, or during, the experiment. In females with ripening ovaries (stages II or III), neither angling treatment significantly reduced standardized gonadal mass. The mean per cent of atretic oocytes increased among females in all groups, but was significantly greater in those that were harshly angled. The results demonstrated that the gonadal development of M. novemaculeata could be suppressed or impaired (by angling, handling and confinement), and that further research is warranted. In particular, the timing and severity of angling in relation to the stage of gonadal development could have important implications for the introduction of temporal restrictions to angling.

The p100 coactivator is present in the nuclei of mammary epithelial cells and its abundance is increased in response to prolactin in culture and in mammary tissue during lactation.

The p100 coactivator, first identified as a coactivator of the Epstein-Barr virus-encoded transcription factor, EBNA-2, in cultured cells, interacts with a number of transcription factors. However, the role of p100 in animals is unclear. We found that the abundance of p100 is closely associated with the lactating state in mammary tissue of mice and cows. Using two antibodies against independent parts of the protein, p100 immunoreactivity was localised to mammary epithelial cells, and was enriched in both nuclei and endoplasmic reticulum/organelle fractions. Stimulation of beta-casein expression in cultured mammary epithelial cells was associated with an increase in abundance of the p100 protein. The relative abundance of p100 mRNA was not altered in mammary tissue throughout the gestation-lactation cycle, indicating that the abundance of p100 is altered by a post-transcriptional mechanism. Further work is required to clarify the function of p100 in mammary epithelial cells.

Stat5 phosphorylation status and DNA-binding activity in the bovine and murine mammary glands.

The transcription factors Stat5a and Stat5b are mediators of prolactin signalling in mammary epithelial cells, and are thought to play a role in lactogenesis. In cultured cells, activation of Stat5 activity through phosphorylation results in Stat5 binding to the promoters of at least some of the milk protein genes, thereby stimulating their transcription. However, the mammary biology of Stat5 differs between species, and the role of Stat5 in the bovine mammary gland is not fully understood. We have generated an antibody that specifically recognises the phosphorylated forms of Stat5a and Stat5b and used it to compare the levels of phosphorylated Stat5 with Stat5 DNA-binding activity in bovine and murine mammary tissue. Both Stat5 DNA-binding activity and phosphorylation status in the bovine mammary gland were at near-maximal levels at late pregnancy (27-35 days prior to calving), when at least three of the major milk proteins are not highly expressed. In addition, these studies revealed significant animal-to-animal variation in the level of Stat5 activity in both species. The results are consistent with a role in terminal differentiation of mammary epithelial cells. They also suggest that the stimulation of high-level expression of milk protein genes in the bovine mammary gland is not through activation of the prolactin receptor-Jak2-Stat5 pathway.

Differences in the abundance of nuclear proteins in the bovine mammary gland throughout the lactation and gestation cycles.

The mechanisms that control the synthesis of milk protein are not fully understood and could well involve undiscovered proteins within the mammary gland. A search for such proteins in high salt extracts of nuclei that had been isolated from bovine mammary tissues was undertaken using two-dimensional electrophoresis on large format gels. The sensitivity of the procedure was sufficient to detect the transcription factors Sp1 and NF-1 by Coomassie blue stain; over 300 proteins were routinely detected. Analysis of mammary tissue taken from 5 nonlactating cows in midpregnancy, 5 cows in late lactation, and 4 cows in early involution revealed five proteins where relative abundance was altered with stage of lactation or reproductive cycle. Four of these proteins were identified by Western blotting or amino acid sequencing as lactoferrin, annexin II, vimentin, and heavy-chain immunoglobulin. Analysis of proteins after further enrichment of the extracts by heparin-Sepharose affinity chromatography revealed an additional protein that was substantially more abundant in samples from lactating cows. This 90-kDa protein did not react with anti-Stat5 antibodies. Conceivably, one or more of these six proteins could play a role in the lactational function of the bovine mammary gland.

Cell-free synthesis of leaf protein: Identification of an apparent precursor of the small subunit of ribulose-1,5-bisphosphate carboxylase.

Cytoplasmic mRNA has been isolated from the leaves of pea seedlings. Translation of this RNA in the wheat germ cell-free system produces two major products, RI and RII, with molecular weights of 33,000 and 20,000, respectively. Both of these products are considerably larger than the small subunit of ribulose-1,5-bisphosphate carboxylase [3-phospho-D-glycerate carboxy-lyase (dimerizing), EC 4.1.1.39], which is the major product of cytoplasmic protein synthesis in vivo and has a molecular weight of 14,000. Antiserum prepared against the small subunit of ribulose-1,5-bisphosphate carboxylase precipitates from the cell-free products, in 2-3% yield, three polypeptides of molecular weights 18,000, 16,000 and 14,000. The smallest of these polypeptides is indistinguishable, by sodium dodecyl sulfate/polyacrylamide gel electrophoresis, from the small subunit of ribulose-1,5-bisphosphate carboxylase. Although the cell-free product RII is not precipitated with antiserum prepared against the small subunit of ribulose-1,5-bisphosphate carboxylase, the two polypeptides do show extensive sequence homology, as indicated by ion exchange chromatography of their tryptic peptides. The production of RII can also be achieved in a polysome-primed cell-free system, where protein synthesis is restricted to the completion of polypeptide chains that have already been initiated in vivo. These results indicate that RII is apparently a precursor of the small subunit of ribulose-1,5-bisphosphate carboxylase. We suggest that the selective transport of cytoplasmically synthesized organelle proteins, like animal secretory proteins, may be achieved via the production of precursor polypeptides.