Development and characterisation of 20 microsatellite loci isolated from the large bent-wing bat, Miniopterus schreibersii (Chiroptera: Miniopteridae) and their cross-taxa utility in the family Miniopteridae

The large bent-wing bat, Miniopterus schreibersii (Kuhl 1819), has a long history of taxonomic uncertainty and many populations are known to be in a state of decline. Microsatellite loci were developed for the taxonomic and population genetic assessment of the Australian complex of this species. Of the 33 primer sets designed for this research, seven (21%) were deemed suitably polymorphic for population-level analyses of the Australian taxa, with five (71%) of these loci revealing moderate to high levels of polymorphism (PIC = 0.56 to 0.91). The cross-taxa utility of the M. schreibersii microsatellite markers was assessed in the microbat (Chiroptera) family Miniopteridae. Sub-species and species covering the Miniopteridae’s global distribution (with the exception of the Middle East) were selected, numbering 25 taxa in total. Amplification was successful for 26 loci, of which 20 (77%) were polymorphic. High cross-taxa utility of markers was observed with amplification achieved for all taxa for between four (20%) and 20 (100%) loci, and polymorphism was considered moderate to high (PIC = 0.47–0.91) for 12 (60%) of these loci. The high cross-taxa utility of the microsatellites reported herein reveal versatile and cost-effective molecular markers, contributing an important genetic resource for the research and conservation of Miniopteridae species worldwide.

Isolation and characterisation via 454 sequencing of microsatellites from the tawny frogmouth, Podargus strigoides (Class Aves, Family Podargidae)

We isolated 24 novel polymorphic microsatellite markers from the tawny frogmouth, a nocturnal bird endemic to Australia, which has successfully adapted to urban environments. Initially, 454 shotgun sequencing was used to identify 733 loci with primers designed. Of these, we trialled 30 in the target species of which all amplified a product of expected size. Subsequently, all 30 of these loci were screened for variation in 25 individuals, from a single population in Melbourne, Victoria, Australia. Twenty-eight loci were polymorphic with observed heterozygosity ranging from 0.03 to 0.96 (mean 0.58) and the number of alleles per locus ranged from 2 to 18 (average of 6.5); we confirmed that 24 loci conformed to Hardy–Weinberg expectations. The 24 loci identified here will be sufficient to unequivocally identify individuals and will be useful in understanding the reproductive ecology, population genetics and the gene flow amongst localities in urban environments where this bird thrives.

Reproductive development, GnRHa-induced spawning and egg quality of wild meagre (Argyrosomus regius) acclimatised to captivity

The objective of the study was to acclimatise wild-caught meagre (Argyrosomus regius) to captivity to produce viable eggs for aquaculture production. Twelve meagre (3 males and 9 females, mean weight = 20 ± 7 kg) were caught and transported to a land-based facility on 26 October 2006. During, March to June 2007, all three males were spermiating and five of the nine females were in vitellogenesis with mean maximum oocyte diameter ≥550 μm. No spontaneous spawning was observed. Two hormone treatments, either a single injection of gonadotropin-releasing hormone agonist (GnRHa, 20 μg kg−1 for females and 10 μg kg−1 for males) or a slow-release implant loaded with the same GnRHa (50 μg kg−1 for females and 25 μg kg−1 for males), were used to induce spawning on three different dates on 26 March 2007, 4 May 2007 and 18 April 2008. From each spawning event, the following parameters were determined: fecundity, number of floating eggs, egg size, fertilisation and hatching success, unfed larval survival, and proximal composition and fatty acid profile of the eggs. In 2007, two females that were injected on 26 March and 4 May spawned a total of 5 times producing 9,019,300 floating eggs and a relative fecundity of 198,200 eggs kg−1 and two different females that were implanted on the same dates spawned 14 times producing 12,430,000 floating eggs and a relative fecundity of 276,200 eggs kg−1. In 2008, a pair that was implanted spawned five times producing a total of 10,211,900 floating eggs and a relative fecundity of 527,380 eggs kg−1. The latency period was 48–72 h. Parameters were compared between hormone treatments, date of hormone induction and parents determined by microsatellites. Percentage hatch and egg size were 70 ± 0.3% and 0.99 ± 0.02 mm, respectively, for GnRHa-implanted fish and were significantly higher (P < 0.05) compared to 30 ± 0.3% and 0.95 ± 0.03 mm, respectively, for injected fish. Few differences were observed in proximal composition and fatty acid profile and for all spawns mean (% dry weight) lipid content was 17.3 ± 3.0%, carbohydrate was 4.4 ± 1.9% and protein was 31.5 ± 6.4% and the essential fatty acids: Arachidonic acid (ARA, 20:4n-6) ranged between 0.9 and 1% (of total fatty acids), eicosapentaenoic acid (EPA 20:5n-3) 7.7–10.4% and docosahexaenoic acid (DHA 22:6n-3), 28.6–35.4%. All good quality spawns were obtained in the second and/or third spawn after GnRHa treatment, whereas all bad quality spawns were obtained either on the first spawn or after the fifth spawn. Both spawning protocols gave commercially viable (1,000,000+) numbers of good quality eggs that could form the basis of a hatchery production.