Protein markers of Marteilia sydneyi infection in Sydney rock oysters, Saccostrea glomerata

Marteilia sydneyi is the causative agent of QX disease in Sydney rock oyster, Saccostrea glomerata . It is responsible for disease outbreaks among oysters that occur during summer and can result in up to 95% mortality. QX disease has significantly decreased S. glomerata production in some areas of Australia's eastern seaboard over the past 30 years. Marteilia sydneyi sporulates in the digestive gland of oysters leading to complete disorganization of the infected tissues. The current study used proteomics to identify potential molecular markers of sporulating M. sydneyi infection during a field trial undertaken in the Georges River, Sydney, between December 2006 and May 2007. Early stages of M. sydneyi infection were detected by polymerase chain reaction, whilst cytological examination was used to identify sporulating M. sydneyi in the gut. Protein expression in oyster haemolymph was assessed during the M. sydneyi infection period by two dimensional electrophoresis. Proteome maps identified significant differences in the expression of four proteins in oysters with sporulating M. sydneyi infections.     

Biochemical and Microbiological Assessment of Crucifix Crab (Charybdis feriatus) Stored at 4°C

Charybdis feriatus is a highly prized crab in the local and international markets of East Asian countries. In local markets, crabs are sold live, iced, or refrigerated at 2°C – 4°C. The present study was carried out to determine the shelf life of whole crab stored at 4°C for 192 h using a set of biochemical, microbiological, and sensorial parameters. Biochemical indicators such as the total volatile base nitrogen (TVB-N) and trimethylamine nitrogen (TMA-N) increased significantly during storage and within an acceptable limit up to 144 h. Regarding the biogenic amines, cadaverine and spermidine increased significantly (p < 0.05) during the storage period. Among them, cadaverine could be proposed as a useful spoilage indicator for this crab species. The mesophilic and psychotropic bacterial count exceeded the limit of acceptance after 144 h of storage. Sensory attributes were gradually diminished with the storage time. Sensory scores were found to correlate well with both mesophiles and psychrotrophs. Based on the results of this study, the meat quality of Charybdis feriatus can be maintained up to 144 h when properly stored at a temperature of 4°C ± 1°C.     

High-temperature-tolerant mungbean (Vigna radiata L.) lines produce better yields when exposed to higher CO2 levels

Increasing global air temperatures, along with rising CO₂ levels, are causing concerns about reducing available freshwater resources and altering cropping patterns. They may influence overall growth and production pattern of crop plants. These likely changes would become major limiting factors for future sustainable food production largely in the tropics and subtropics. Thus, understanding physiological responses hold the key to determining the functional relationship between the environment and crop performance. We explore here the impact of rising CO₂ on the growth and yield traits of a few selected high-temperature (HT)-tolerant mungbean lines, which we earlier screened for HT tolerance using a physiological assay under managed growth conditions. The HT-tolerant lines grown under elevated CO₂ levels (550 and 700 μL L^?1) showed a considerable improvement in growth rates (13.5%, 67.8%, and 46.5% in plant height, leaf area, and total dry matter, respectively) and pod and seed yield (48.7% and 31.7%, respectively), compared to local checks under the same environments. Interestingly, the symptoms of accelerated pod maturity were also observed in most of these lines. The outcome of the study would undoubtedly open up opportunities for increased yield potentials of legumes under the conditions of the warming climate and elevated levels of carbon dioxide.     

Unilateral scrotal pyocele in ram caused by Staphylococcus capitis

A massive unilateral scrotal pyocele caused by Staphylococcus capitis in a 6‐year‐old ram is reported. Ultrasound examination of the right hemiscrotum showed an irregular hyperechoic mass in an anechoic fluid. A dense exudate was collected from the scrotum for microbiological analysis. Grossly, there was an extensive greenish purulent exudate and a completely atrophied right testis. Coagulase‐negative S. capitis was isolated in pure culture. To the authors' knowledge, this is the first report of genital infection by S. capitis in rams. This microorganism should be included in the differential diagnosis of ovine genital infections.     

Cloning and characterisation of the ahpA gene of Pasteurella multocida serogroup b:2 (strain P52): short communication

Pasteurella multocida B:2 is responsible for haemorrhagic septicaemia in cattle and buffaloes, causing severe economic losses in the developing countries. In the present study, the ahpA gene of P. multocida B:2 (P52) was cloned, sequenced and compared with the previously reported ahpA gene sequence in P. multocida A:1, which is responsible for its haemolytic phenotype. E. coli DH5a cells were further transformed with recombinant plasmid carrying the ahpA gene from P. multocida B:2 (P52) but SDS-PAGE analysis failed to show the expression of haemolysin protein. Slight haemolysis was albeit observed in horse blood agar plates streaked with recombinant E. coli carrying the ahpA gene. Our study indicates that there is 99.6% similarity and 0.4% divergence between ahpA gene of P. multocida B:2 (P52) and P. multocida A: 1, while membrane topology analysis has predicted that ahpA is an inner membrane protein with two strong hydrophobic regions at the N and C terminals. The presence of significant homology in ahpA sequence in A: 1 and B:2 perhaps suggests a common mechanism of pathogenesis in different species of animals.     

Active root distribution pattern of <Emphasis Type="Italic">Hevea brasiliensis</Emphasis> determined by radioassay of latex serum

The active root distribution pattern of mature rubber (Hevea brasiliensis Muell. Arg.) up to a lateral distance of 250 cm from the tree and to a soil depth of 90 cm was studied in an oxisol by employing ³²P soil injection technique in Kerala, the state which accounts for 83% of rubber cultivation in India. The trees were aged 18 years and grown at a spacing of 4.9 × 4.9 m. The extent of absorption of applied ³²P by the tree from various placements was assessed by radio assay of leaf and latex serum. Latex serum registered higher counts and variability was less compared to leaf indicating the suitability of latex serum as a potential source for radio assay for ³²P studies in rubber. The results revealed that rubber is a surface feeder with 55% of the root activity confining to the top 10 cm of soil layer. Root activity declined with increasing depths and the concentration of physiologically active roots at 90 cm depth was only 6%. A more or less uniform distribution of root activity was noticed with respect to lateral distance indicating more extensive spread of lateral roots. Concentration of physiologically active roots in the surface layer suggests the possibility for competition under intercropped situation in mature plantations.     

Aroma volatiles emission in relation to chilling injury in ‘Kensington Pride’ mango fruit

Summary

Mature green mango fruit (Mangifera indica L. ‘Kensington Pride’) were stored at 0, 5, 10, 15 or 20°C for 1, 3, 7, 14, 21 or 28 d to induce different levels of chilling injury (CI) and to elucidate its relationship with aroma volatile production. The fruit were removed from storage and allowed to ripen at 22 ± 1°C and CI index was assessed on fully ripe fruit. Aroma volatile compounds were estimated from the pulp of fully ripe fruit. CI index significantly increased as the storage temperature was lowered and the storage period was extended, particularly in the fruit stored at 0, 5 or 10°C. CI symptoms did not develop on fruit stored at 15 or 20°C. Fifty-six aroma volatile compounds were identified from mango fruit pulp using headspace solid phase microextraction (SPME) technique with gas chromatography (GC-FID) and GC combined with mass spectrometry (GC-MS). Among the 56 aroma volatile compounds, 25 were quantified using GC, which included monoterpenes (±-pinene, ²-pinene, myrcene, 2-carene, ±-phellandrene, 3-carene, ±-terpinene, limonene, ocimine, ³-terpinene, ±-terpinolene, and a-terpineol), sesquiterpenes (±-copaene, ±-gurjunene, trans-carophyllene, aromadendrane, ±-humulene, alloaromadendrane, ³-gurjunene, and ledene), hydrocarbon (p-cymene), esters (methyl octanoate and ethyl caprylate), aldehyde (decanal), and norisoprenoid (²-ionone). A significant reduction in total aroma volatiles, monoterpenes, sesquiterpenes, hydrocarbon, esters, aldehyde, and norisoprenoid production were observed in fruit stored at 0, 5, 10 or 15°C compared with fruit stored at 20°C. The degree of reduction in aroma volatile compounds depended on the severity of CI, induced with different storage temperatures and storage periods. In conclusion lower temperature storage induced CI in mango fruit and adversely affected the aroma volatiles production.     

Location choices of undocumented migrants: Does access to higher public education matter?

We examine the impact of differences in state‐level higher education policies on location choices of undocumented immigrants in the U.S. using six years of available data over the period 2000 to 2012. The effect of greater access to higher education on the percentage of undocumented migrants residing in a state is positive, suggesting the existence of a “higher public education magnet” effect. Among states with favorable higher education policies, undocumented migrants prefer those states with larger networks, despite the likelihood of greater competition for admission to public institutions of higher education. However, undocumented migrants are reluctant to locate in states that have both large networks and unfavorable educational policies. This is possibly because the fear of attracting additional restrictive regulations dominates the positive cost‐saving effect of large networks.     

Protein Composition of Seminal Plasma in Fractionated Stallion Ejaculates

Seminal plasma (SP) contains several types of compounds derived from the epididymides and accessory glands. The aim of this study was to examine the protein composition of different ejaculate fractions. Trial I: fractionated ejaculates were collected from two normal and two subfertile stallions. Samples containing pre‐sperm fluid and the first sperm‐rich jets (HIGH‐1), the main sperm‐rich portion (HIGH‐2), the jets with low sperm concentrations (LOW), and a combined whole‐ejaculate (WE) sample was centrifuged, and the SP was filtered and frozen. A part of each SP sample was stored (5°C, 24 h) with spermatozoa from HIGH‐2 and skim milk extender. Sperm motility was evaluated after storage in extender mixed with the stallion’s own SP or SP from one of the other stallions (sperm from a normal stallion stored in SP from a subfertile stallion and vice versa). Protein composition was analysed using reverse‐phase liquid chromatography (RP‐HPLC), N‐terminal sequencing and mass spectrometry. The area‐under‐the‐curve (AUC) was used for quantitative comparison of proteins within fractions. Trial II: semen samples were collected from seven stallions. Fractions with the highest (HIGH) and lowest (LOW) sperm concentrations and WE samples were examined using SDS‐PAGE and densitometry. No significant differences emerged between fractions in the AUC‐values of the Horse Seminal Protein‐1 (HSP‐1) and HSP‐2 peaks, or the peak containing HSP‐3 and HSP‐4 (HSP‐3/4). Levels of HSP‐1, HSP‐2 and HSP‐3/4 were not significantly correlated with total sperm motility, progressive sperm motility or average path velocity after storage. Significant differences between ejaculate fractions in the amount of different protein groups present in SP were not found in Trial I; but in Trial II, the proteins in the 60–70 kDa range were more abundant in LOW than in HIGH and WE, indicating that this band contained proteins derived mainly from the seminal vesicles, which produce most of the SP in LOW.     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.