Evaluation of otorhinolaryngologic, audiologic, and genetic findings in children with cystic fibrosis: A tertiary care experience.

OBJECTIVES: To evaluate otorhinolaryngologic findings and the relationship between aminoglycoside (AG) exposure and hearing loss in paediatric patients with cystic fibrosis (cwCF). We also aimed to investigate the genetic predisposition to AG ototoxicity by screening for m.1555A>G mutations. METHODS: CwCF who underwent otorhinolaryngologic and audiologic examinations were retrospectively included. Clinical characteristics, ear-nose-throat related symptoms, and a history of ototoxic drug exposure were recorded. m.1555A>G mutations were retrospectively screened among patients with audiologic evaluations. RESULTS: Two hundred thirty-four cwCF were included in this study with a median age of 10.7 (range, 6.8-14.2) years. Nasal obstruction (14.1%) was the most common symptom. Fifty-two (22.2%) patients had chronic rhinosinusitis (CRS) with nasal polyps (CRSwNP). There was a positive correlation between CRSwNP and the symptom of nasal obstruction (r:.234, p < .001), snoring (r:.179, p = .006), and sleeping with mouth open (r:.138, p = .034). One hundred forty-nine (63.6%) patients had audiologic evaluations; 14 (9.4%) had hearing impairment. No statistical significance existed between ototoxicity and IV AG exposure (p = .90). Six (42.8%) of 14 patients did not receive ototoxic drugs. One hundred nineteen (50.8%) patients were screened for m.1555A>G mutations, and none were detected. CONCLUSIONS: Almost a quarter of the study population had CRSwNP. Neither the relationship between AGs exposure and hearing loss nor the genetic predisposition to AG ototoxicity could be shown in cwCF.

Multiple species of canine Rhipicephalus complex detected in Canada.

Multiple species of brown dog ticks have been described in the United States and the Caribbean: Rhipicephalus sanguineus sensu stricto (s.s.), also referred to as temperate lineage; R. linnaei (=tropical lineage); and R. rutilus (=southeastern Europe lineage) However, Rhipicephalus spp. are rarely recovered from dogs in Canada. To identify canine Rhipicephalus spp. in Canada and determine the influence of travel history on infestation, ticks morphologically identified as brown dog ticks (n = 93) collected from dogs (n = 13) in British Columbia, Ontario, and Québec, Canada were submitted with information regarding each dog's geographic location and travel history. Nucleic acid was extracted from available individual ticks (n = 86) and PCR was used to amplify sequences of a 12S rRNA mitochondrial gene fragment. Sequences were compared to published reference sequences of known species and a phylogenetic tree constructed. Twenty-three ticks (26.7%) consistent with R. linnaei were identified on seven dogs, including dogs from British Columbia and Ontario, with a median infestation intensity of 2 ticks/dog (mean = 3.3 ticks/dog). Sixty-one ticks (70.9%) consistent with R. sanguineus s.s. were found on two dogs from Québec and Ontario (median = 30.5 ticks/dog; mean = 30.5 ticks/dog). One dog from Ontario was infested with R. rutilus (n = 2) (2.3%). Species could not be determined for ticks from three dogs from Ontario and Québec. Most infested dogs (10/13; 76.9%) had a recent (< 1 month) international travel history. These data confirm that multiple species of canine Rhipicephalus are occasionally found in Canada and suggest introduction following travel is likely responsible for these infestations. Further analysis will allow for greater understanding of the range and diversity of canine Rhipicephalus spp. in North America and may reveal risk factors for infestation.

On the way between Africa and Europe: Molecular taxonomy of ticks collected from birds in Malta.

The Maltese Archipelago is situated in the middle of the Mediterranean Basin, between Europe and Africa, therefore representing an important stopover site for migratory birds between these two continents. Despite this, up-to-date information is not available on tick species associated with birds in Malta. Therefore, in this study, birds mist-netted for ringing by BirdLife Malta were examined for the presence of ticks between September, 2019 and May, 2021. Ticks were identified morphologically and molecularly, using three genetic markers. During the study period, 57 individuals of 22 bird species were found tick-infested, from which altogether 113 ixodid ticks were collected. The majority of developmental stages were nymphs, but 13 larvae and one female were also found. These ticks belonged to nine species: Ixodes cumulatimpunctatus (n=1), Ixodes ricinus (n=2), Ixodes acuminatus (n=2), Ixodes frontalis (n=5), Ixodes festai (n=1), one species of the Amblyomma marmoreum complex (n=8), Hyalomma rufipes (n=78), Hyalomma marginatum (n=7) and Hyalomma lusitanicum (n=1). Eight Hyalomma sp. ticks could only be identified on the genus level. Regarding seasonality, all Palearctic Ixodes species were carried by birds exclusively in the autumn (i.e., north to south), whereas H. rufipes (with predominantly Afrotropical distribution) was exclusively collected in the spring (i.e., carried south to north). Two tick species that occurred on birds in Malta, i.e., a species of the A. marmoreum complex and I. cumulatimpunctatus are only indigenous in the Afrotropical zoogeographic region. This is the first finding of the latter tick species in Europe, and four tick species were identified for the first time in Malta. In conclusion, the diversity of tick species regularly arriving in Europe from Africa is most likely higher than reflected by data obtained in Mediterranean countries of mainland Europe. Most notably, ticks of the genus Amblyomma appear to be underrepresented in previous datasets. Ticks of the subgenus Afrixodes (represented by I. cumulatimpunctatus) might also be imported into Europe by migratory birds.

Single nucleotide polymorphism-based methodology for authentication of bovine, caprine, ovine, camel, and donkey meat cuts.

To guarantee food safetyand sustainability, it is necessary to verify meat authenticity. This study focused on the development of single nucleotide polymorphism-based polymerase chain reaction-restriction fragment length polymorphism (SNP-based PCR-RFLP) and forensically informative nucleotide sequence (FINS) methodologies based on PCR amplification of the mitochondrial 12S rRNA gene for discrimination of six red meat species, that is, cattle, buffalo, goat, sheep, camel, and donkey. FINS allowed the unambiguous identification of all species analyzed. In addition, six SNPs, where a restriction site for TasI could be localized using a preliminary in silico analysis, gave a unique RFLP pattern for each species. The results revealed a low level of species substitution (8%) in the tested meat samples. In particular, one buffalo and goat samples have been substituted with cow and sheep, respectively. Finally, the developed techniques herein showed high potentials to be routinely used as reliable and fast tools to avoid meat species substitutions. PRACTICAL APPLICATION: This research deals with genetic techniques to trace meats. This kind of research helps the concerned agencies to build capacity to safeguard consumer sentiments as well as providing better market access and better food price and quality for the consumer.

Markers of Genetic Variation in Blue Gourami (Trichogaster trichopterus) as a Model for Labyrinth Fish.

Markers of genetic variation between species are important for both applied and basic research. Here, various genes of the blue gourami (Trichogaster trichopterus, suborder Anabantoidei, a model labyrinth fish), many of them involved in growth and reproduction, are reviewed as markers of genetic variation. The genes encoding the following hormones are described: kisspeptins 1 and 2, gonadotropin-releasing hormones 1, 2, and 3, growth hormone, somatolactin, prolactin, follicle- stimulating hormone and luteinizing hormone, as well as mitochondrial genes encoding cytochrome b and 12S rRNA. Genetic markers in blue gourami, representing the suborder Anabantoidei, differ from those in other bony fishes. The sequence of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene of blue gourami is often used to study the Anabantoidei suborder. Among the genes involved in controlling growth and reproduction, the most suitable genetic markers for distinguishing between species of the Anabantoidei have functions in the hypothalamic-pituitary-somatotropic axis: pituitary adenylate cyclase-activating polypeptide and growth hormone, and the 12S rRNA gene.

Molecular characterization of Echinococcus granulosus in livestock of Al-Madinah (Saudi Arabia).

Echinococcus granulosus is the causative agent of cystic echinococcosis, which has serious impacts on human and/or animal health, resulting in significant economic losses. Echinococcus granulosus comprises a number of intra-specific variants or strains at the genetic level. In Saudi Arabia, few studies were performed on genetic variations in Echinococcus species. Therefore, the present study aimed to investigate the phenotypic and genetic characterization of hydatid cysts harboured by sheep and camels in Al-Madinah Al-Munawarah. Samples of hydatid cysts were collected from local sheep (n = 25) and camels (n = 8). The morphological criteria of protoscoleces were investigated. To investigate the molecular characterization, random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR), single-stranded conformation polymorphism (SSCP) were carried out. DNA was extracted from individual fertile cysts and subjected to RAPD-PCR analysis (using five arbitrary primers) and PCR amplification of cytochrome c oxidase I (cox1) and 12S ribosomal ribonucleic acid (12S rRNA) genes. The PCR products were subjected to SSCP analysis for genetic discrimination in E. granulosus isolates. In addition, partially sequencing of the mitochondrial DNA cox1 genes was achieved for assessing the phylogenetic positions of collected isolates using some global published sequence data of cox1 genes. The rostellar hooks of camel and local sheep isolates show remarkable variability in their dimensions. Five distinct SSCP patterns were identified in the 12S rRNA gene, showing intraspecific variations in E. granulosus of camels and local sheep. Sequencing of (cox1) genes of both local sheep and camels exhibit high similarity with those of the same gene (E. granulosus sensu stricto) published in NCBI BLAST.

Molecular identification of Taenia hydatigena and Mesocestoides species based on copro-DNA analysis of wild carnivores in Mongolia.

Cyclophyllidean tapeworms obligatorily parasitize numerous mammalian species, including herbivores, domestic animals and humans, of which, the genera Taenia and Mesocestoides are well characterized. However, little is known about these parasitic infections in wild animals. This study aims to investigate the prevalence and distribution of Taenia sp. and Mesocestoides sp. in wild carnivores in Mongolia by identifying tapeworm species based on mtDNA analysis. The field survey was carried out in 2012-2013 in 19 provinces located in different ecological regions. A total of 405 fecal samples from wild carnivores were collected. Specific DNA markers in fecal samples was detected via copro-DNA analysis and tapeworm species were identified by DNA sequencing. From 27.7% (112/405) of samples, cox1 and 12S rRNA genes of tapeworms were amplified. Further, Taenia hydatigena (50.0%, 56/112) and two Mesocestoides species, including Mesocestoides sp.-1 (36.6%, 41/112) and Mesocestoides sp.-2 (13.4%, 15/112) were identified by DNA sequencing. The prevalence of T. hydatigena was 19.9% (27/136), 13.8% (23/167), 4.8% (3/62), and 7.5% (3/40) in wolves, red foxes, corsac foxes, and snow leopards, respectively. The prevalence of Mesocestoides sp.-1 was 14.7% (20/136), 9% (15/167), 9.7% (6/62) in wolves, red foxes, and corsac foxes, while the prevalence of Mesocestoides sp.-2 was 4.4% (6/136), 1.8% (3/167), 3.2% (2/62), and 10.0% (4/40) in wolves, red foxes, corsac foxes, and snow leopards, respectively. T. hydatigena was found throughout all ecological regions, while Mesocestoides sp.-1 was in the mountain taiga, forest-steppe, steppe, desert-steppe, and desert, and Mesocestoides sp.-2 in the alpine, forest-steppe, steppe, and desert-steppe ecoregions. This study revealed the prevalence and distribution of cyclophyllidean tapeworms in wild carnivores in Mongolia; while also confirming that wolves, red foxes, corsac foxes, and snow leopards serve as definitive hosts for unidentified Mesocestoides species.

Detection and Quantification of Milk Ingredients as Hidden Allergens in Meat Products by a Novel Specific Real-Time PCR Method.

Milk ingredients are often included in a wide range of meat products, such as cooked hams and sausages, to improve technological characteristics. However, milk proteins are also important food allergens. The aim of this study was the development of a highly sensitive and specific real-time PCR system targeting the 12S rRNA gene of Bos domesticus for the detection and quantification of milk as an allergenic ingredient in processed meat products. The method was able to achieve an absolute limit of detection (LOD) of 6 fg of milk DNA. Using a normalized approach (∆Ct method) for the detection of milk protein concentrate (MPC), it was possible to obtain sensitivities down to 0.01% (w/w) of MPC in model hams (raw and cooked) and autoclaved sausages, and 0.005% in raw sausage mixtures. The developed systems generally presented acceptable PCR performance parameters, being successfully validated with blind samples, applied to commercial samples, and further compared with an immunochemical assay. Trace amounts of milk material were quantified in two out of 13 samples, but the results mostly infer the excessive practice of the precautionary labeling.

Altered mitochondrial DNA methylation and mitochondrial DNA copy number in an APP/PS1 transgenic mouse model of Alzheimer disease.

Alzheimer's disease (AD) is a chronic neurodegenerative disease and mitochondrial impairment is a key feature of AD. The mitochondrial DNA (mtDNA) epigenetic mechanism is a relatively new field compared to nuclear DNA. The relationship between mtDNA epigenetic mechanism and AD hasn't been established. So we analyzed the mtDNA methylation in D-loop region and 12 S rRNA gene in the hippocampi in amyloid precursor protein/presenilin 1 (APP/PS1) transgenic mice by bisulfite pyrosequencing. Mitochondrial DNA copy number and gene expression were studied by quantitative real-time PCR (qRT-PCR). We observed a decrease in the displacement loop (D-loop) methylation and an increase in 12 S rRNA gene methylation, while both the mtDNA copy number and the mitochondrial gene expression were reduced in APP/PS1 transgenic mice. In summary, the present finding suggest that mtDNA methylation may play a role in AD pathology, which warrants larger future investigations.

Detection of fraud by addition cow's milk in cheese buffalo and its connection with seasonality / Detecção de fraude por adição de leite de vaca em queijo de búfala e a relação com a sazonalidade

The seasons influence the production of buffalos’ milk. Because of this, the producers may produce a mixture of buffalo and bovine milk during cheese production in periods of low production. Therefore, the present work aimed to investigate fraud in buffalo cheese and the relationship between seasonality and different physicochemical properties of buffalo cheeses produced and marketed in eastern Amazonia. We obtained commercial samples of buffalo cheese during two Amazonian climatic periods from commercial points of Marajó-Pará, Brazil. After collection, there were lipid, protein, ash, and humidity analyses. Determination of carbohydrates and energy values was also performed for the nutritional characterization of samples, as well as for mPCR analysis to detect buffalo and/or bovine DNA. DNA extraction protocol of the samples was standardized and two pairs were used for the mPCR reaction, amplifying fragments of approximately 220 bp for Bubalus bubalis DNA and 346 bp fragments for Bos taurus DNA. Among the samples acquired in the rainy season, we observed that 33% were inadequately labeled, indicating fraud from cow’s milk incorporation and fraud from substitution of raw material. From the nine samples obtained in the dry season, all the samples showed cow’s milk incorporation fraud. The highest fraud rate coincided with the period of low milk production from buffalo and there was a difference in composition between fraudulent and non-fraudulent cheeses. Therefore, seasonality influences increase in cattle milk for the production of buffalo cheese, and this adulteration may decrease the nutritional content of the product.

As estações climáticas influenciam a produção de leite de búfala. Isso pode levar os produtores a misturarem os leites de búfala e bovino durante a produção de queijo em períodos de baixa produção. Portanto, o presente trabalho teve como objetivo verificar fraudes em queijo de búfala, a relação com a sazonalidade e as diferenças físico-químicas de queijos de origem bubalina, produzidos e comercializados no leste da Amazônia. Foram coletadas amostras comerciais de queijo de búfala em dois períodos climáticos da Amazônia em pontos comerciais do Marajó-Pará, Brasil. Após a coleta foram realizadas análises de lipídios, proteínas, cinzas e umidade. A determinação dos carboidratos e do valor energético também foi feita para a caracterização nutricional das amostras, bem como a análise de mPCR para a detecção de DNA de búfalo e/ou bovino. Para isso, padronizou-se um protocolo de extração de DNA das amostras e utilizou-se dois pares na reação mPCR, amplificar fragmentos de aproximadamente 220 pb para o DNA de Bubalus bubalis e fragmentos de 346 pb para o Bos taurus. Entre as amostras adquiridas na estação chuvosa, observou-se que 33% foram rotuladas inadequadamente, caracterizando fraude por incorporação de leite de vaca e fraude por substituição de matéria-prima. Das 9 amostras coletadas no período seco, todas as amostras apresentaram fraude na incorporação do leite de vaca. Este estudo revelou que a maior taxa de fraude coincide com o período de baixa produção de leite e que há uma diferença na composição entre queijos fraudulentos e não fraudulentos. Portanto, a sazonalidade influencia no acréscimo de leite de bovinos na produção de queijo de búfala e que esta adulteração pode diminuir o conteúdo nutricional do produto.

Detection of fraud by addition cow's milk in cheese buffalo and its connection with seasonality / Detecção de fraude por adição de leite de vaca em queijo de búfala e a relação com a sazonalidade

The seasons influence the production of buffalos' milk. Because of this, the producers may produce a mixture of buffalo and bovine milk during cheese production in periods of low production. Therefore, the present work aimed to investigate fraud in buffalo cheese and the relationship between seasonality and different physicochemical properties of buffalo cheeses produced and marketed in eastern Amazonia. We obtained commercial samples of buffalo cheese during two Amazonian climatic periods from commercial points of Marajó-Pará, Brazil. After collection, there were lipid, protein, ash, and humidity analyses. Determination of carbohydrates and energy values was also performed for the nutritional characterization of samples, as well as for mPCR analysis to detect buffalo and/or bovine DNA. DNA extraction protocol of the samples was standardized and two pairs were used for the mPCR reaction, amplifying fragments of approximately 220 bp for Bubalus bubalis DNA and 346 bp fragments for Bos taurus DNA. Among the samples acquired in the rainy season, we observed that 33% were inadequately labeled, indicating fraud from cow's milk incorporation and fraud from substitution of raw material. From the nine samples obtained in the dry season, all the samples showed cow's milk incorporation fraud. The highest fraud rate coincided with the period of low milk production from buffalo and there was a difference in composition between fraudulent and non-fraudulent cheeses. Therefore, seasonality influences increase in cattle milk for the production of buffalo cheese, and this adulteration may decrease the nutritional content of the product.

As estações climáticas influenciam a produção de leite de búfala. Isso pode levar os produtores a misturarem os leites de búfala e bovino durante a produção de queijo em períodos de baixa produção. Portanto, o presente trabalho teve como objetivo verificar fraudes em queijo de búfala, a relação com a sazonalidade e as diferenças físico-químicas de queijos de origem bubalina, produzidos e comercializados no leste da Amazônia. Foram coletadas amostras comerciais de queijo de búfala em dois períodos climáticos da Amazônia em pontos comerciais do Marajó-Pará, Brasil. Após a coleta foram realizadas análises de lipídios, proteínas, cinzas e umidade. A determinação dos carboidratos e do valor energético também foi feita para a caracterização nutricional das amostras, bem como a análise de mPCR para a detecção de DNA de búfalo e/ou bovino. Para isso, padronizou-se um protocolo de extração de DNA das amostras e utilizou-se dois pares na reação mPCR, amplificar fragmentos de aproximadamente 220 pb para o DNA de Bubalus bubalis e fragmentos de 346 pb para o Bos taurus. Entre as amostras adquiridas na estação chuvosa, observou-se que 33% foram rotuladas inadequadamente, caracterizando fraude por incorporação de leite de vaca e fraude por substituição de matéria-prima. Das 9 amostras coletadas no período seco, todas as amostras apresentaram fraude na incorporação do leite de vaca. Este estudo revelou que a maior taxa de fraude coincide com o período de baixa produção de leite e que há uma diferença na composição entre queijos fraudulentos e não fraudulentos. Portanto, a sazonalidade influencia no acréscimo de leite de bovinos na produção de queijo de búfala e que esta adulteração pode diminuir o conteúdo nutricional do produto.

A group-specific, quantitative real-time PCR assay for detection of crab, a crustacean shellfish allergen, in complex food matrices.

A real-time PCR assay was developed for detection of crab, a crustacean allergen, in food products. Group-specific primers and probes were developed to detect numerous species of crab. Method validation included tests of detection in complex food matrices, evaluation of commercial food products, and cross-reactivity testing on a wide variety of crustaceans. The method was able to detect several species of crab spiked into complex food matrices at levels ranging from 0.1 to 105 parts per million (weight/weight), worked equally well on different platforms, exhibited high specificity for crab over other types of crustaceans, and yielded much higher signals from commercial food products listing crab as an ingredient than from those containing other crustaceans.

The high resolution melting analysis (HRM) as a molecular tool for monitoring parasites of the wildlife.

In an interconnected world, the international pet trade on wild animals is becoming increasingly important. As a consequence, non-native parasite species are introduced, which affect the health of wildlife and contribute to the loss of biodiversity. Because the investigation of parasite diversity within vulnerable host species implies the molecular identification of large samples of parasite eggs, the sequencing of DNA barcodes is time-consuming and costly. Thereby, the objectives of our study were to apply the high resolution melting (HRM) approach for species determination from pools of parasite eggs. Molecular assays were validated on flatworm parasites (polystomes) infecting the Mediterranean pond turtle Mauremys leprosa and the invasive red-eared slider Trachemys scripta elegans in French natural environments. HRM analysis results indicated that double or multiple parasitic infections could be detected from wild animal populations. They also showed that the cycle of parasite eggs production was not regular over time and may depend on several factors, among which the ecological niche and the target species. Thereby, monitoring parasites from wild endangered animals implies periodic parasitological surveys to avoid false negative diagnostics, based solely on eggs production.

Analysis of deafness-associated mutant genes and pedigrees in neonates / 预防医学

Objective To carry out mutation analysis of deafness-associated genes for deaf newborns and their parents, and to estimate the recurrence risk for their parents to have deaf descendants.Methods Suspected cases of inherited deafness were identified by neonatal hearing screening and questionnaires. Genomic DNAs of suspected cases and their parents were extracted from their peripheral blood samples . Common deafness-associated genes(i.e. GJB2,SLC26A4 and 12S rRNA genes)were amplified by polymerase chain reaction(PCR),and those PCR products were sequenced for the mutation analysis.Results From 2013 to 2016, 193 cases of deafness were found in neonatal hearing screening,29 cases of suspected as hereditary deafness were screened,and 17 out of 29 cases were found to have mutations in deafness-associated genes(detection rate:58.62%). GJB2 homozygous mutations were identified in two cases and their parents,and the recurrence risk to have deaf descendants was 100%. Four cases of suspected hereditary deafness had GJB2 homozygous mutations,and their parents were both GJB2 mutation carriers. There was one case with SLC26A4 homozygous mutations,and their parents were both SLC26A4 mutation carrier. Two cases were detected to have GJB2 V371 homozygous mutations,and their parents were both GJB2 V371 mutation carriers. For those seven parents carrying deafness-associated mutations above,the recurrence risk of deafness for their descendants was 25%.Conclusion In addition to hearing screening,the genetic diagnosis of deafness-associated genes is helpful to clarify the cause of suspected neonatal hereditary deafness,and can provide objective reproductive counseling and guidance for those deaf parents or parents with deaf children.

Levels of genetic diversity and taxonomic status of Epinephelus species in United Arab Emirates fish markets.

Understanding the patterns of genetic diversity of fish species is essential for marine conservation and management. This is particularly important in the Arabian Gulf where marine life is subject to extreme environmental conditions that could impact genetic diversity. Here we assess genetic diversity of the most commercially important fish in the United Arab Emirates; groupers (Epinephelus spp.). Sequencing of 973 bp mitochondrial DNA from 140 tissue samples collected in four main fish markets revealed 58 haplotypes clustered within three groups. Data analysis revealed the presence of three distinct Epinephelus species being marketed as one species (hammour): Epinephelus coioides, Epinephelus areolatus and Epinephelus bleekeri. We report species-specific genetic markers and demonstrate that all three species exhibit relatively low levels of genetic variation, reflecting the effect of overfishing and environmental pressures. In light of the genetic evidence presented here, conservation and management of groupers in the UAE warrant the implementation of species-specific measures.

Sequence variability in the mitochondrial 12S rRNA and tRNA Val genes of Ixodes scapularis (Acari: Ixodidae) individuals shown previously to be genetically invariant.

The DNA sequences of the mitochondrial (mt) 12S rRNA and tRNA(Val) genes were characterized for 82 blacklegged ticks (Ixodes scapularis) that were genetically identical for Domains IV and V of the mt 16S rRNA gene. Thirty-one haplotypes, differed in sequence by 1-9 bp, were detected among the 82 ticks. Most nucleotide alterations in DNA sequence did not affect the stability of the secondary structures of the RNAs. The magnitude of the DNA sequence variation in the mt 12S rRNA and tRNA(Val) genes among blacklegged ticks suggests that this region of the mitochondrial genome has potential as a genetic marker for examining the population genetics and phylogeography of I. scapularis.

An effective strategy for species identification of avian meats using the mitochondrial 12S rRNA gene fragment.

An effective DNA-based molecular method had been used to identify avian species from meats. The method combined the use of a pair of universal primers, which amplified about 440-bp fragment of the mitochondrial 12S rRNA gene. A total of 99 meat samples were tested and 17 haplotypes were identified by DNA sequencing, which representing 14 avian species. One avian species was listed as the national first-grade protected animal in China and the IUCN endangered species. Two avian species were under the national second-grade state protection. The proposed method represents a straightforward and robust method for the accurate identification of avian species that could be used by law enforcement agencies as a tool for the control of illegal trade of meat from protected species.

Normal hearing in a child with the m.1555A>G mutation despite repeated exposure to aminoglycosides. Has the penetrance of this pharmacogenetic interaction been overestimated?

The mtDNA m.1555A>G mutation causes increased susceptibility to aminoglycoside ototoxicity resulting in significant hearing loss in 100% of reported exposed cases. Genetic and audiological assessments were conducted in a sample of 59 children with cystic fibrosis (CF) undergoing aminoglycoside treatment. Of the two m.1555G patients identified one had severe-profound deafness. Surprisingly, the second m.1555G patient exhibited well-preserved hearing despite repeated exposure. This may be a rare case of intact hearing in an m.1555G individual with aminoglycoside use. Alternatively, its penetrance may have been previously overestimated due to recruitment bias. Further studies are required to determine the true penetrance to inform m.1555A>G genetic testing in similar clinical scenarios.

Avian-specific real-time PCR assay for authenticity control in farm animal feeds and pet foods.

A highly sensitive TaqMan real-time PCR assay targeting the mitochondrial 12S rRNA gene was developed for detection of an avian-specific DNA fragment (68bp) in farm animal and pet feeds. The specificity of the assay was verified against a wide representation of animal and plant species. Applicability assessment of the avian real-time PCR was conducted through representative analysis of two types of compound feeds: industrial farm animal feeds (n=60) subjected to extreme temperatures, and commercial dog and cat feeds (n=210). Results obtained demonstrated the suitability of the real-time PCR assay to detect the presence of low percentages of highly processed avian material in the feed samples analysed. Although quantification results were well reproducible under the experimental conditions tested, an accurate estimation of the target content in feeds is impossible in practice. Nevertheless, the method may be useful as an alternative tool for traceability purposes within the framework of feed control.