Periodontal disease in free-ranging koalas (Phascolarctos cinereus) from the Mount Lofty Ranges, South Australia, and its association with koala retrovirus infection.

BACKGROUND: In northern Australian koala populations (Queensland and New South Wales), periodontal disease (gingivitis and periodontitis) is common while koala retrovirus subtype A is endogenous, with other subtypes transmitted exogenously. Koala retrovirus has been hypothesised to cause immune suppression and may predispose koalas to diseases caused by concurrent infections. In southern Australia populations (Victoria and South Australia) periodontal disease has not been investigated, and koala retrovirus is presumably exogenously transmitted. This study described oral health in South Australian koalas and investigated if an association between periodontal disease and koala retrovirus exists. METHODS: Oral health was examined for wild-caught koalas from the Mount Lofty Ranges (n = 75). Koala retrovirus provirus was detected in whole blood using nested PCR and proviral load determined with qPCR. Periodontal disease severity was recorded and used to calculate the Final Oral Health Index (0-normal, 24-severe).Results Periodontal disease was observed in 84% (63/75) of koalas; 77% had gingivitis (58/75) and 65% (49/75) had periodontitis. The average Final Oral Health Index was 5.47 (s.d 3.13). Most cases of periodontal disease were associated with the incisors. Koala retrovirus-infected koalas were more likely to present with periodontitis (p = 0.042) and the Final Oral Health Index was negatively correlated with proviral load (ρ = -0.353, p = 0.017). CONCLUSION: South Australian koalas had a high prevalence of gingivitis and periodontitis. Periodontal disease was more prevalent in the incisors. Exogenous koala retrovirus infection may also facilitate the development of periodontitis by modulation of the immune response to concurrent oral bacterial infections.

Oxalate-degrading bacteria, including Oxalobacter formigenes, colonise the gastrointestinal tract of healthy koalas (Phascolarctos cinereus) and those with oxalate nephrosis.

BACKGROUND: Koalas in the Mount Lofty Ranges, South Australia, have a high prevalence of oxalate nephrosis, or calcium oxalate kidney crystals. Gastrointestinal tract oxalate-degrading bacteria, particularly Oxalobacter formigenes, have been identified in other animal species and humans, and their absence or low abundance is postulated to increase the risk of renal oxalate diseases. This study aimed to identify oxalate-degrading bacteria in the gastrointestinal tract of koalas and determine their association with oxalate nephrosis. METHODS: Caecal and faecal samples were collected at necropsy from 22 Mount Lofty Ranges koalas that had been euthanased on welfare grounds, with 8 koalas found to have oxalate nephrosis by renal histopathology. Samples were analysed by PCR for the oxc gene, which encodes oxalyl-CoA decarboxylase, and also by Illumina sequencing of the V3-V4 region of the bacterial 16S rRNA gene. RESULTS: The oxc gene was detected in 100% of koala samples, regardless of oxalate nephrosis status. Oxalobacter formigenes was detected in all but one faecal sample, with no difference in abundance between koalas affected and unaffected by oxalate nephrosis. Other species of known oxalate-degrading bacteria were infrequently detected. CONCLUSION: This is the first study to identify Oxalobacter and other oxalate-degrading bacterial species in koalas, but an association with oxalate nephrosis and absence or low abundance of Oxalobacter was not found. This suggests other mechanisms underlie the risk of oxalate nephrosis in koalas.

New insights into the expression profile of MicroRNA-34c and P53 in infertile men spermatozoa and testicular tissue.

Spermatogenesis is proliferation and differentiation processes of stem spermatogonia into mature spermatozoa controlled by the genes responsible for transcription and post transcription levels. MicroRNAs (miRNA) are  the key factors during gene expression in RNA silencing and post-transcriptional regulation. They play main roles in regulation of early and late spermatogenesis, and reproduction. In this study, we investigate the role of miRNAs in infertile males.The patients were assigned to five groups based on semen analysis (n=55), including normozoospermic (N), moderate oligoasthenoteratozoospermic (MOAT), severe oligoasthenoteratozoospermic (SOAT), obstructive azoospermia (OA) and non-obstructive azoospermia (NOA). Quantitative RT-PCR was recruited to study the expression of miR-34c and tumor suppressor p53 gene. In addition, malondialdehyde (MDA) and DNA fragmentation was measured. Network analysis was performed using Pathway Studio web tool (Elsevier). Our results revealed statistically significant increased expression of miR-34c in moderate oligoasthenoteratozoospermic, non-obstructive azoospermia and an increased expression of p53 in MOAT, SOAT and NOA males. Also, the percentage of DNA fragmentation and oxidative stress was significantly higher in infertile groups (MOAT and SOAT) than other groups. These findings provide a novel molecular mechanism of gene regulation during cell-cycle and apoptosis in sperm, which gives a new regulatory insight into male infertility in terms of molecular diagnosis.

Differential expression of TLP, ERF1, and R2R3MYB in annual Medicago species under salinity conditions.

The present study was conducted to evaluate the responses of three annual Medicago species (M. truncatula, M. laciniata, and M. polymorpha) to salinity. We analyzed publicly available microarray data in NCBI pertaining to salinity-response genes in M. truncatula. Our data search identified Tubby C2 (TLP) and ethylene responsive transcription factor 1 (ERF1) as genes that potentially respond to salinity. We evaluated morpho-physiological traits and the expression of the genes in three Medicago species that had been maintained under control and saline conditions. The analysis of morpho-physiological traits showed that M. polymorpha and M. laciniata were more tolerant to salinity than M. truncatula, as they had lower reductions in biomass and dry root weight and lower increases in anthocyanin concentration. The saline conditions caused a significant increase (P < 0.01) in the expression of TLP in all Medicago species, but caused a significant decrease in the expression of ERF1. Considerable variation in anthocyanin concentrations was found among the three Medicago species. To investigate the cause of this variation, we examined the expression of R2R3MYB, a gene involved in the biosynthesis of anthocyanins. Our analysis showed that saline conditions induced high over-expression of R2R3MYB in all three Medicago spp. The high efficiency of the primer pairs used in qRT-PCR enabled us to compare the expression levels of each gene in the three species. We concluded that the more salt tolerant species showed higher expression of TLP and R2R3MYB under both control and salinity conditions.

Isolation and in silico functional analysis of MtATP6, a 6-kDa subunit of mitochondrial F1F0-ATP synthase, in response to abiotic stress.

Mitochondrial F(1)F(0)-ATP synthase is a key enzymatic complex of energy metabolism that provides ATP for the cell. Subunits of this enzyme over-express under stress conditions. Little is known about the structure and regulatory mechanism of the F(0) portion of this enzyme. We isolated the full-length coding sequence of the RMtATP6 gene from rice and wheat, and partial sequences from Aegilops crassa and Triticum monococcum (Poaceae). We found that the sequence of rice RMtATP6 is 1965 bp long and contains two exons and one intron in 3'-UTR. Then, we analyzed the 2000-bp upstream region of the initiation codon ATG of the RMtATP6 and AtMtATP6, as promoter. The RMtATP6 coding sequence was found to be much conserved in the different plant species, possibly because of its key role under stress conditions. Promoter analysis demonstrated that RMtATP6 and AtMtATP6 include cis-acting elements such as ABRE, MYC/MYB, GT element in the upstream region, which respond to abscisic acid stress hormone and might show vital its roles in biotic and abiotic tolerance as an early-stress responsive gene. A mitochondrial signal peptide of 30 amino acids in length and an N-terminal cleavage site between amino acids 20 and 21 were discovered in RMtATP6. In addition, we found a transmembrane domain with an alpha helix structure that possibly passed through the mitochondrial inner membrane and established the 6-kDa subunit in the F(0) portion of the enzyme complex. Apparently, under stress conditions, with increasing ATP consumption by the cell, the 6-kDa subunit accumulates; by switching on F(1)F(0)-ATP synthase it provides additional energy needed for cell homeostasis.

Application of functional genomic information to develop efficient EST-SSRs for the chicken (Gallus gallus).

Many years of domestication and breeding have given rise to the wide range of chicken breeds that exist today; however, an increasing number of local chicken breeds are under threat of extinction. A comprehensive characterization of chicken markers (especially type I markers) is needed to monitor and conserve genetic diversity in this species. The explosion of genomics and functional genomics information in recent years has opened new possibilities for the generation of molecular markers. We analyzed a large number of expressed sequence tags (ESTs) to test the possibility of using EST-derived microsatellite markers for investigating the Gallus gallus genome. Chromosomal locations for the majority of these SSRs were predicted. Of the 31,576 unigenes assembled from the 544,150 redundant EST sequences, 1757 SSR markers were discovered on 1544 ESTs, using the SSRLocator software, with an average density of 28.7 kb per SSR. The dimer motifs were the most frequent (46.38%), followed by trimeric (38.58%), tetrameric (10.19%), pentameric (4.5%), and hexameric (<1%) markers. Different from the case for cattle and sheep, AT/TA was the most abundant dimeric repeat, accounting for 41.71% of all dimeric repeats in the chicken ESTs. The EST-SSR distribution was not uniform among the chromosomes; the majority of the EST-SSRs were located on chromosomes GGA2 and GGA10. We found that most of the EST-SSRs are involved in positive regulation of cellular and metabolic processes. This is the first time that EST sequences have been mined to find chicken microsatellites. On average, 3.8% of the G. gallus UniGene sequences could be exploited for development of EST-SSRs, indicating a good source for molecular markers as well as for functional genome analysis.

Amino Acid Features of P1B-ATPase Heavy Metal Transporters Enabling Small Numbers of Organisms to Cope with Heavy Metal Pollution.

Phytoremediation refers to the use of plants for extraction and detoxification of pollutants, providing a new and powerful weapon against a polluted environment. In some plants, such as Thlaspi spp, heavy metal ATPases are involved in overall metal ion homeostasis and hyperaccumulation. P1B-ATPases pump a wide range of cations, especially heavy metals, across membranes against their electrochemical gradients. Determination of the protein characteristics of P1B-ATPases in hyperaccumulator plants provides a new opportuntity for engineering of phytoremediating plants. In this study, using diverse weighting and modeling approaches, 2644 protein characteristics of primary, secondary, and tertiary structures of P1B-ATPases in hyperaccumulator and nonhyperaccumulator plants were extracted and compared to identify differences between proteins in hyperaccumulator and nonhyperaccumulator pumps. Although the protein characteristics were variable in their weighting, tree and rule induction models; glycine count, frequency of glutamine-valine, and valine-phenylalanine count were the most important attributes highlighted by 10, five, and four models, respectively. In addition, a precise model was built to discriminate P1B-ATPases in different organisms based on their structural protein features. Moreover, reliable models for prediction of the hyperaccumulating activity of unknown P1B-ATPase pumps were developed. Uncovering important structural features of hyperaccumulator pumps in this study has provided the knowledge required for future modification and engineering of these pumps by techniques such as site-directed mutagenesis.