Molecular expression and identification of caprine estrogen receptor gene 1 for fertility status in bucks

Estrogen and its receptors are essential for sexual development and reproduction. Oestrogen receptor alpha (ERα) is a nuclear receptor activated by the hormone oestrogen. In male, ERα is encoded by the gene ESR1 (oestrogen receptor1) responsible for better fertility. ESR1 is involved in the reabsorption of luminal fluid during the transit of spermatozoa from the testis to the head of the epididymis which is important for their survival and maturation during epididymal storage. The absence of ESR1 leads to reduced epididymal sperm content, reduced sperm motility and fertilizing ability. The present study was undertaken to investigate the expression and presence of ESR1 gene in fertile and low-fertile male goat breeds. We identified ESR1 gene through various molecular tools. Genotyping was carried out by high resonance melting analysis using Roche Light Cycler 480(LC-480) system and found three different genotypes. Genotypic frequency-AA (blue-0.67), BB(Red-0.2), AB(Green-0.08) with allele frequency A(0.71 and B (0.29). The predominance of this gene in head of epididymis in fertile bucks was confirmed by SDS-PAGE, Western blotting and immunohistochemistry. From the results, we corroborated that the present study provides a useful and effective way to predict male fertility in goat breeds, which in turn increases the percentage of fertility in flock leading to more number of offspring in a kidding season.     

Development of two cell culture systems from Asian seabass Lates calcarifer (Bloch)

Two new cell culture systems namely epitheloid cells of Lates (LCE) and fibroblastic cells of Lates (LCF) have been developed from fry and fingerling of the economically important brackishwater fish Lates calcarifer. Primary cultures were initiated by explant technique using caudal fin of fingerling and whole body tissue of the fry. The nutritional requirements and the growth pattern in response to different culture environment were similar for the two cell cultures. The culture medium used was Leibovitz‐15 supplemented with 20% fetal bovine serum (FBS) and 1% fish serum. The LCE comprised of epithelioid cells and LCF cells were fibroblastic. With a split ratio of 1:2, the confluency of cells was attained in 8–10 days at an incubation temperature of 28°C. The cells were found to grow well in a wide range of temperature (24–32°C) and stable at 20 and 36°C. The growth rate of LCF and LCE cells increased proportionately with the concentration of FBS from 5% to 20%. A decrease of serum level to 10% after eight subcultures produced no apparent change in cell morphology and growth rate. The viability of cells was found to be 70% when revived after a month of storage in liquid nitrogen (?196°C).     

Influence of Benzyladenine on Leaf Senescence and Photosynthesis in Sunflower (Helianthus annuus L.)

The effect of foliar application of benzyladenine on sunflower leaf senescence, photosynthesis, RuBP carboxylase activity and diffusion resistance was investigated. A considerable drop in photosynthetic rate and rise in diffusion resistance were noted dunng the seed filling period. Benzyladenine application maintained the chlorophyll content and chlorophyll a : b ratio at higher level, enhanced RuBP carboxylase activity and net photosynthesis in both upper and lower leaves. A lower level of diffusion resistance was also observed by benzyladenine application during seed filling period. The results suggested that application of benzyladenine at 30, 60 and 90 days after sowing could help to maintain photosynthesis and assimilate supply to the developing seeds of sunflower.     

Clinical skills and knowledge used by veterinary graduates during their first year in small animal practice

Objective To determine the clinical skills and areas of knowledge used by veterinarians in small animal practice during their first year after graduation and the degree of assistance and supervision they received while developing these skills. Design A postal survey was sent in December 1999 to 59 veterinarians who completed their training at Murdoch University in December 1998. Procedure The first part of the survey asked for information on veterinary work patterns since leaving university. The second part consisted of a list of diagnostic and therapeutic skills of varying complexity and the graduates were asked to indicate whether they had used these skills in practice and whether they had been assisted or supervised while doing them. The respondents were also asked if they had had the opportunity to practise these skills as undergraduates at university or during extramural experience. In the third part the areas of knowledge used in practice were assessed by analysis of a series of consecutive cases. Results Forty replies were received but since three graduates had done no small animal work the analysis of the skills section is based on 37 responses. Thirty graduates supplied information on 994 canine cases and 308 feline cases. The distribution of the mean work time was dogs and cats 69%, horses 13%, farm animals 11%, birds 3% and others 4%. Skills used by over 90% of graduates included general anaesthesia, examination of the tympanic membrane, taking and interpreting an abdominal radiograph, catheterising a male cat, fine needle aspiration of a mass, neutering dogs and cats, tooth scaling and extraction and treating an aural haematoma. The survey also identified the opportunities for undergraduates to practise some of these skills during extramural experience and the extent of assistance given to new graduates during their first year in practice. The areas of knowledge used in over 10% of the cases included vaccination, anaesthesia/sedation, skin/coat problems, general advice on pet health, neutering and musculoskeletal diseases. Conclusion Veterinarians, in their first year after graduation, use a wide range of complex diagnostic and therapeutic skills. Although many of these skills are acquired during the undergraduate training, a significant contribution is made by extramural practical work undertaken during the clinical years of the undergraduate course and in the first year following graduation. Practising veterinarians play an important role in providing opportunities and supervision for clinical training.     

A national serological survey to verify Australia's freedom from porcine reproductive and respiratory syndrome

Objective To provide serological data to support Australia's claim of freedom from porcine reproductive and respiratory syndrome.
Design A national serological survey was designed to provide 99% confidence of detecting at least one infected pig herd in Australia, assuming that at least 5% of herds would have been exposed to porcine reproductive and respiratory syndrome virus and that at least 25% of the 'finisher' pigs in these herds would have antibodies to the virus.
Procedure A two-stage testing regime was used. All samples were tested with a commercially available enzyme-linked immunosorbent assay. If assay reactions were found, all samples from the herd were to be tested using the indirect immunofluorescence antibody assay.
Results Of the 875 samples from 163 herds from all States in Australia, there was some evidence of reactivity in only four samples from four herds on the enzyme-linked immunosorbent assay. Further testing using the indirect immunofluorescence antibody assay according to the study protocol demonstrated that the reactions were not due to the presence of specific porcine reproductive and respiratory syndrome virus antibodies in the sera.
Conclusion The results of this study support the view that Australian pigs are free of porcine reproductive and respiratory syndrome virus.     

The Fusarium graminearum genome reveals a link between localized polymorphism and pathogen specialization

We sequenced and annotated the genome of the filamentous fungus Fusarium graminearum, a major pathogen of cultivated cereals. Very few repetitive sequences were detected, and the process of repeat-induced point mutation, in which duplicated sequences are subject to extensive mutation, may partially account for the reduced repeat content and apparent low number of paralogous (ancestrally duplicated) genes. A second strain of F. graminearum contained more than 10,000 single-nucleotide polymorphisms, which were frequently located near telomeres and within other discrete chromosomal segments. Many highly polymorphic regions contained sets of genes implicated in plant-fungus interactions and were unusually divergent, with higher rates of recombination. These regions of genome innovation may result from selection due to interactions of F. graminearum with its plant hosts.     

Evapotranspiration losses for pepper under plastic mulch and shallow water table conditions

Use of literature crop coefficient (K _c) values for quantifying evapotranspiration (ET_c) under non-standard conditions such as plastic mulch, shallow water table, and sub-tropical conditions can lead to inaccurate ET_c estimates. A 5-year experiment was conducted for fall crop growing seasons in south Florida to quantity bi-weekly ET_c and K _c for bell pepper grown under shallow water table and plastic mulch environments using large drainage lysimeters. The ET_c values varied from 205 to 320 mm with a seasonal average of 267 mm. Average K _c values for bell pepper for development, mid-season, and late stages were 1.05, 1.21, and 1.28, respectively. Higher than literature initial K _c values were due to rainfall and use of sub-irrigation system to maintain artificially high water table which results in high soil moisture in the bare soil area—such high moisture results in high evaporation. The K _c values from this study were statistically higher than literature values. Use of literature K _c values resulted in underestimating ET_c by 27–37%. The K _c values would provide improved estimates of sub-irrigated pepper ET_c in subtropical Florida and elsewhere with similar environment.     

Standardization of Rootstocks of Mango (Mangifera Indica L.)

Mango cuttings could be induced to form roots provided IBA at 5,000 p.p.m. in lanolin was applied at the base of shoots after removing a ring of bark while they were still attached to the mother tree.

Cuttings taken from one-month-old seedlings gave the highest percentage of rooting and establishment, but with increasing age of the mother plants the rooting was considerably reduced. The rooting of cuttings of different age groups could be improved by using shoots forced into vigorous growth by hard pruning. Younger plants responded better to such treatment than older ones.

Cuttings from etiolated shoots rooted better than those from non-etiolated, but etiolation was less effective in promoting rooting than invigoration by hard pruning. Etiolation and invigoration combined gave the highest percentage of rooting and establishment in all age groups.