Regional Workshop on Co-management in Small-Scale Fisheries: Lessons Learned and Best Practices 12-13 December 2012, Bangkok, Thailand

An FAO project, the Regional Fisheries Livelihoods Programme, which is funded by Spain, has worked with the Timorese authorities and coastal communities to build local capacity and put in place effective methods to gather a variety of important fisheries data. This is used to help make important decisions relating to the management of the nation's fisheries sector. These actions, which are detailed in this publication, have been carried out at relatively little expense and in a participatory manner that has engaged communities while at the same time providing practical skills to all involved

Restriction Fragment Length Polymorphism Linkage Map for Arabidopsis thaliana

We have constructed a restriction fragment length polymorphism linkage map for the nuclear genome of the flowering plant Arabidopsis thaliana. The map, containing 90 randomly distributed molecular markers, is physically very dense; >50% of the genome is within 1.9 centimorgans, or approx 270 kilobase pairs, of the mapped DNA fragments. The map was based on the meiotic segregation of markers in two different crosses. The restriction fragment length polymorphism linkage groups were integrated with the five classically mapped linkage groups by virtue of mapped mutations included in these crosses. Markers consist of both cloned Arabidopsis genes and random low-copy-number genomic DNA clones that are able to detect polymorphisms with the restriction enzymes EcoRI, Bgl II, and/or Xba I. These cloned markers can serve as starting points for chromosome walking, allowing for the isolation of Arabidopsis genes of known map location. The restriction fragment length polymorphism map also can associate clones of unknown gene function with mutant phenotypes, and vice versa

Genetic diversity among Frankia strains nodulating members of the family Casuarinaceae in Australia revealed by PCR and restriction fragment length polymorphism analysis with crushed root nodules

DNA extracted directly from nodules was used to assess the genetic diversity of #Frankia# strains symbiotically associated with two species of genus #Casuarina# and two of the genus #Allocasuarina# naturally occuring in northeastern Australia. DNA from field-collected nodules or extracted from reference cultures of #Casuarina#-infective #Frankia# strains was used as the template in PCRs with primers targeting two DNA regions, one in the ribosomal operon and the other in the #nif# operon. PCR products were then analysed by using a set of restriction andonucleases. Five distinct genetic groups were recognized on the basis of these restriction patterns. These groups were consistently associated with the host species from which the nodules originated. All isolated reference strains had similar patterns and were assigned to group 1 along with six of eight unisolated #Frankia# strains from #Casuarina equisetifolia# in Australia. Group 2 consisted of two unisolated #Frankia# strains from #C. equisetifolia#, whereas groups 3 to 5 comprised all unisolated strains from #Casuarina cunninghamiana#, #Allocasuarina torulosa#, and #Allocasuarina littoralis#, respectively. These results demonstrate that, contrary to the results previous molecular studies of isolated strains, there is genetic diversity among #Frankia# strains that infect members of the family Casuarinaceae. The apparent high homogeneity of #Frankia# strains in these previous studies probably relates to the single host species from which the strains were obtained and the origin of these strains from areas outside the natural geographic range of members of the family Casuarinaceae, where genetic diversity could be lower than in Australia. (Résumé d'auteur

Prevalence, antimicrobial resistance and genetic diversity of Campylobacter coli and Campylobacter jejuni in Ecuadorian broilers at slaughter age

Thermotolerant Campylobacter spp. are a major cause of foodborne gastrointestinal infections worldwide. The linkage of human campylobacteriosis and poultry has been widely described. In this study we aimed to investigate the prevalence, antimicrobial resistance and genetic diversity of C. coli and C. jejuni in broilers from Ecuador. Caecal content from 379 randomly selected broiler batches originating from 115 farms were collected from 6 slaughterhouses located in the province of Pichincha during 1 year. Microbiological isolation was performed by direct plating on mCCDA agar. Identification of Campylobacter species was done by PCR. Minimum inhibitory concentration (MIC) values for gentamicin, ciprofloxacin, nalidixic acid, tetracycline, streptomycin, and erythromycin were obtained. Genetic variation was assessed by RFLP-flaA typing and Multilocus Sequence Typing (MLST) of selected isolates. Prevalence at batch level was 64.1%. Of the positive batches 68.7% were positive for C. coli, 18.9% for C. jejuni, and 12.4% for C. coli and C. jejuni. Resistance rates above 67% were shown for tetracycline, ciprofloxacin, and nalidixic acid. The resistance pattern tetracycline, ciprofloxin, and nalidixic acid was the dominant one in both Campylobacter species. RFLP-flaA typing analysis showed that C. coli and C. jejuni strains belonged to 38 and 26 profiles respectively. On the other hand MLST typing revealed that C. coli except one strain belonged to CC-828, while C. jejuni except 2 strains belonged to 12 assigned clonal complexes (CCs). Furthermore 4 new sequence types (STs) for both species were described, whereby 2 new STs for C. coli were based on new allele sequences. Further research is necessary to estimate the impact of the slaughter of Campylobacter positive broiler batches on the contamination level of carcasses in slaughterhouses and at retail in Ecuador

Characterization of Ascaris from Ecuador and Zanzibar

To shed light on the epidemiology of ascariasis in Ecuador and Zanzibar, 177 adult worms retrieved by chemo-expulsion from either people or pigs were collected, measured and subjected to polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the ribosomal internal transcribed spacer (ITS) region. Upon double digestion with RsaI and HaeIII, PCR-RFLP analysis revealed the presence of A. lumbricoides in people and A. suum in pigs in Ecuador. In contrast, while there are no pigs on Zanzibar, of the 56 worms obtained from people, one was genotyped as A. suum. No additional genetic variation was detected upon further PCR-RFLP analysis with several other restriction enzymes. Upon measurement, worm mass and length differed by location and by species, A. suum being lighter and longer. While there is no evidence to suggest zoonotic transmission in Ecuador, an enduring historical signature of previous zoonotic transmission remains on Zanzibar

Allelic Frequency of Kappa-Casein Locus (Asp148/Ala) in F1: Simmental (Bos Taurus) X Ongole Grade (Bos Indicus)

This study was conducted to detect the genetic variants (single nucleotide polymorphism) of kappa-casein locus (Asp148/Ala) in F1: Simmental (Bos taurus) x Ongole grade (Bos indicus), SIMPO. Genomic DNA was isolated from blood sample of 40 SIMPO (21 males and 19 females). A 780 bp specific fragment of kappa-casein gene spanning from the forth exon region (517 bp) to forth intron (263 bp) was successfully amplified. The result of the PCR-RFLP (Polymerase Chain Reaction - Restriction Fragment Length Polymorphisms) analysis using HindIII enzyme showed that two genotypes (AA and AB) were found at this locus in SIMPO. The frequencies of A and B alleles in SIMPO were 0.79 and 0.21, respectively. The frequency lies between B. taurus (Simmental) and B. indicus group

Association of Calpastatin (CAST) Gene with Growth Traits and Carcass Characteristics in Bali Cattle

Calpastatin (CAST) gene is well known as an inhibitor of muscle protein degradation and relates to muscle growth and meat tenderness. The objective of this study was to determine the association of CAST gene with growth traits and carcass characteristics in Bali cattle. A number of data from 35 Bali bulls were collected from BPTU Bali Cattle to obtain growth traits, carcass characteristics, and blood samples. Polymorphism of CAST gene in Bali bulls was analyzed by using PCR-RFLP and DNA sequencing. The association of CAST gene with growth traits and carcass characteristics were analyzed by using General Linear Model (GLM). The result showed that there were two genotypes (GG and AG) of CAST gene with allele frequencies of 0.857 and 0.143, respectively, for G and A. Notably, mutation A to G occurred in 253 bp CAST fragment gene in Bali Cattle. Genotypes GG and AG of CAST gene significantly affected (P<0.05) the back-fat thickness and longissimus dorsi without a significant effect on the growth traits. It could be concluded that CAST gene had a potency as a marker gene for carcass quality in Bali cattle

Nitrogen removal and ammonia-oxidising bacteria in a vertical flow constructed wetland treating inorganic wastewater

Nitrogen removal performance and the ammonia-oxidising bacterial (AOB) community were assessed in the batch loaded 1.3ha saturated surface vertical flow wetland at CSBP Ltd, a fertiliser and chemical manufacturer located in Kwinana, Western Australia. From September 2008 to October 2009 water quality was monitored and sediment samples collected for bacterial analyses. During the period of study the wetland received an average inflow of 1109m3/day with NH3-N = 40mg/L and NO3-N=23mg/L. Effluent NH3-N and NO3-N were on average 31mg/L and 25mg/L respectively. The overall NH3-N removal rate for the period was 1.2g/m2/d indicating the nitrifying capacity of the wetland. The structure of the AOB community was analysed using group specific primers for the ammonia monooxygenase gene (amoA) by terminal restriction fragment length polymorphism (T-RFLP) and by clone libraries to identify key members. The majority of sequences obtained were most similar to Nitrosomonas sp. while Nitrosospira sp. was less frequent. Another two vertical flow wetlands, 0.8ha each, were commissioned at CSBP in July 2009, since then the wetland in this study has received nitrified effluent from these two new cells

DNA sequence evidence for speciation, paraphyly and a Mesozoic dispersal of cancellothyridid articulate brachiopods

Because the classification of extant and fossil articulate brachiopods is based largely upon shell characters observable in fossils, it identifies morphotaxa whose biological status can, in practice, best be inferred from estimates of genetic divergence. Allozyme polymorphism and restriction fragment length polymorphism of mitochondrial DNA (mtDNA RFLP) have been used to show that nuclear and mitochondrial genetic divergence between samples of the cancellothyridid brachiopods Terebratulina septentrionalis from Canada and T. retusa from Europe is compatible with biological speciation, but the genetic distances obtained were biased by methodological limitations. Here, we report estimates of divergence in 12S rDNA mitochondrial sequences within and between samples of these brachiopods. The sequence-based genetic distance between these samples (5.98-0.07% SE) is at least 10 times greater than within them and, since they also differ in a complex life-history trait, their species status is considered to be securely established. Divergence levels between 12S rDNA genes of three other cancellothyridids, T. unguicula from Alaska, T. crossei from near Japan, and Cancellothyris hedleyi from near Australia are higher than between the two North Atlantic species, and the mean nucleotide distance between all these cancellothyrids is similar to the mean distance between species of Littorina (Mollusca: Gastropoda). Sequences of both 12S and 16S genes from cancellothyridids and other short-looped brachiopod species show neither saturation nor lineage-specific rate differences and, when analysed with different outgroups, either separately or together, yield one unexpected, but well-supported, tree with Alaskan T. unguicula basal and C. hedleyi nested within Terebratulina, i.e. these genera are paraphyletic. A geologically dated divergence between Antarctic and New Zealand species of the short-looped brachiopod Liothyrella is used to calibrate the rate of 12S divergence at ca. 0.1% per million years (MY), and this rate is used to infer that T. septentrionalis and T. retusa have been diverging for ca. 60 MY and that they and T. unguicula have been diverging from their last common ancestor for ca. 100 MY. This indicates a Mesozoic origin for the present-day distribution of cancellothyridids and the basal position of T. unguicula suggests a possible North Pacific centre of origin, with separate Atlantic and Pacific radiations. The inclusion of Cancellothyris within Terebratulina also shows that adult shell characters such as umbo, foramen and symphytium shape, whilst probably indispensible for the practical classification of fossils, are not reliable guides to genealogy

Symbiont diversity is not involved in depth acclimation in the Mediterranean sea whip Eunicella singularis

In symbiotic cnidarians, acclimation to depth and lower irradiance can involve physiological changes in the photosynthetic dinoflagellate endosymbiont, such as increased chlorophyll content, or qualitative modifications in the symbiont population in favour of better adapted strains. It has been argued that a lack of capacity to acquire new symbionts could limit the bathymetric distribution of the host species, or compromise its long-term survival in a changing environment. But is that always true? To address this question, we investigated the symbiont genetic diversity in Eunicella singularis, a Mediterranean sea whip species with a wide bathymetric distribution (10 to 50 m depth), which has recently suffered from mass mortalities after periods of abnormally high sea temperatures. We measured symbiont population densities and chlorophyll content in natural populations, and followed the response of the holobionts after reciprocal transplantations to deep and shallow depths. A total of 161 colonies were sampled at 2 depths (10 and 30 m) at 5 sites in the northwestern Mediterranean. All colonies harboured a single ribosomal Symbiodinium clade (A'), but a relatively high within-clade genetic diversity was found among and within colonies. This diversity was not structured by depth, even though the deeper colonies contained significantly lower population densities of symbionts and less chlorophyll. We did, however, reveal host-symbiont specificity among E. singularis and other Mediterranean cnidarian species. Transplantation experiments revealed a limit of plasticity for symbiont population density and chlorophyll content, which in turn questions the importance of the trophic role of Symbiodinium in E. singularis