Polymerase chain reaction amplification of 16S-23S spacer region for rapid identification of Salmonella serovars

Polymerase chain reaction (PCR) was used to amplify the spacer regions between the 16S and 23S genes of rRNA genetic loci of Salmonella serovars for their rapid identification. These genetic loci revealed a significant level of polymorphism in length across the species/serovar lines. When the 16S-23S spacer region amplification products were subjected to agarose electrophoresis, the patterns observed could be used to distinguish all the serovars of Salmonella tested. Unique elements obtained in amplification products were mostly clustered at serovar level, although certain genus-specific patterns were also observed. On the basis of the results obtained, the amplification of 16S-23S ribosomal spacer region could suitably be used in a PCR-based identification method for Salmonella serovars.     

Impact of Endocrine‐Disrupting Chemicals on Reproductive Function in Zebrafish (Danio rerio)

The prevalence of endocrine‐disrupting chemicals (EDCs) in the aquatic environment has been associated with the wide detection of alterations in the development and physiology of vertebrates. Zebrafish, as a model species, has been extensively used in toxicological research. In this review, we focus on recent published evidence of the harmful effects of EDCs on reproductive function in zebrafish, including skewed sex ratio, immature gonads, diminished sexual behaviour, decreased sperm count, reduced spawning and fertilization. These impairments mostly result from disruption to sex‐steroid hormones induced by endocrine disruptors. We also discuss other effects of exposure to EDCs. In EDC exposure research, despite incomplete assessments of altered gonad histopathology and sexual behaviour, these present potential effective biomarkers or pathways for evaluating the reproductive function in zebrafish on EDC exposure. To date, the pernicious effects of some EDCs on the reproductive performance in laboratory zebrafish are well understood; however, similar alterations remain for further determination in wild‐type fish and more kinds of EDCs. More studies should be performed under established scientific regulatory criteria to investigate the impact of EDCs on reproduction in zebrafish. Moreover, further research is required to explain the definite mechanism of sexual differentiation, which helps in understanding the shift of sexual phenotype with EDC exposure.     

A note on canopy and air temperature equality

Well-watered crops having nearly identical ratio of intercept and slope of the “non-water-stressed baseline” would attain their equivalence point temperature at roughly identical air vapor pressure deficit. However, such a behavior of these crops might not prevail as the wet soil dries out progressively. The methods for calculating the equivalence point temperature due to Linacre (1964) and Paw U (1984) gave results which sometimes differed by more than 10 K.     

Detection of goat pox antigen and antibody by the counter immunoelectrophoresis test

Summary The counterimmunoelectrophoresis (CIE) test was standardised for the detection of goat pox antigen and antibody using inactivated antigens. The chloroform inactivated and live antigens were equally sensitive for detection of goat pox precipitins. The precipitinogens of goat pox virus (GPV) were found to be soluble in nature. The CIE test was quick as well as more sensitive than the agar gel precipitation test for detection of GPV antibody/antigen. The CIE employing inactivated antigen has been used for the first time in the detection of GPV antibodies/antigens.

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Deteccion Del Antigeno Y Anticuerpos De Viruela Caprina Mediate La Prueba De Contrainmunoelectroforesis

Resumen Se estandarizó la prueba de contrainmunoelectroforesis, para la detección del antígeno y anticuerpos del virus de la viruela caprina, usando antígenos inactivados. El antígeno inactivado con cloroformo y el antígeno vivo, fueron igualmente sensitivos para la detección de precipitinas de viruela caprina. Los precipitógenos del virus de la viruela caprina se encontraron que eran solubles. La prueba de contrainmunoelectroforesis fue más rápida y más sensitiva que la precipitación agar gelatina para la detección de anticuerpos/antígenos del virus de la viruela caprina. La prueba de contrainmunoelectroforesis con antígeno inactivado ha sido utilizada por vez primera en la detección de anticuerpos/antígenos del virus de la viruela caprina.

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Detection De l'Antigene Et De l'Anticorps Variole Caprine Par Un Test De Contrimmuno-Electrophorese

Résumé Le test de contrimmuno-électrophorèse (CIE) a été standardisé pour la détection de l'antigène et de l'anticorps variole caprine avec des antigènes inactivés. Les antigènes vivants et inactivés par le chloroforme sont de sensibilité équivalente pour la détection des précipitines variole caprine. On a montré que ces précipitogènes du virus variole caprine (VVC) étaient de nature soluble. Le CIE est rapide et plus sensible que le test de précipitation en gélose pour la détection des antigènes et anticorps VVC. C'est la première fois que le CIE mettant en oeuvre un antigène inactivé a été utilisé.

     

Mapping and marker-assisted breeding of a gene allelic to the major Asian rice gall midge resistance gene <Emphasis Type="Italic">Gm8</Emphasis>

Host plant resistance is the preferred management strategy for Asian rice gall midge (Orseolia oryzae), a serious pest in many rice-growing countries. Identification of simple sequence repeat (SSR) markers that are tightly linked to pest resistance genes can accelerate development of gene pyramids for durable/multiple resistance. Based on conventional and molecular allelism tests, we report herein that rice genotype Aganni possesses Gm8 gene, conferring hypersensitive independent (HR– type) resistance to gall midge biotypes GMB1, GMB2, GMB3, GMB4, and GMB4M. The gene Gm8 was mapped to chromosome 8 within a 400-kbp region, and the SSR markers RM22685 and RM22709 flank the gene closely. Using these closely linked flanking markers, nine other gall midge-resistant genotypes were identified as carrying the same gene Gm8. Through marker-assisted selection, Gm8 has been introgressed into an elite bacterial blight-resistant cultivar, Improved Samba-Mahsuri (IS).     

Assessment of Carbon Balance in Community Forests in Dolakha, Nepal

This study assessed the net above-ground carbon stock in six community forests in the Dolakha district, Nepal. A survey was conducted of above-ground timber species, using random sampling. A tree-ring chronology for Pinus roxburghii was created to construct a growth model representative of the various mainly-pine species. The allometric model combined with tree ring analysis was used to estimate carbon stock and annual growth in the above-ground tree biomass. The out-take of forest biomass for construction material and fuelwood was estimated on the basis of interviews and official records of community forest user groups. The average annual carbon increment of the community forests was 2.19 ton/ha, and the average annual carbon out-take of timber and fuelwood was 0.25 ton/ha. The net average carbon balance of 1.94 ton/ha was equivalent to 117.44 tons of carbon per community forest annually. All the community forests were actively managed leading to a sustainable forest institution, which acts as a carbon sink. It is concluded that community forests have the potential to reduce emissions by avoiding deforestation and forest degradation, enhance forest carbon sink and improve livelihoods for local communities.     

Mucopolysaccharidosis type VI in a Miniature Poodle-type dog caused by a deletion in the arylsulphatase B gene

Abstract

AIM: To investigate and characterise an inborn error of metabolism in a dog with skeletal and ocular abnormalities.

METHODS: A 2.5-year-old small male Miniature Poodle-like dog was presented with gross joint laxity and bilateral corneal opacities. Clinical examination was augmented by routine haematology, serum chemistry, radiographs, pathology, enzymology and molecular genetic studies. Euthanasia was requested when the dog was 3 years of age because of progressively decreasing quality of life.

RESULTS: Radiology revealed generalised epiphyseal dysplasia, malformed vertebral bodies, luxation/subluxation of appendicular and lumbosacral joints with hypoplasia of the odontoid process and hyoid apparatus. These clinical and radiographic findings, together with a positive urinary Berry spot test for mucopolysaccharides, and metachromatic granules in leucocytes, were indicative of a mucopolysaccharidosis (MPS), a lysosomal storage disease. Histological lesions included vacuolation of stromal cells of the cornea, fibroblasts, chondrocytes, macrophages and renal cells. The brain was essentially normal except for moderate secondary Wallerian-type degeneration in motor and sensory tracts of the hind brain. Dermatan sulphate-uria was present and enzymology revealed negligible activity of N-acetylgalactosamine-4-sulphatase, also known as arylsulphatase B, in cultured fibroblasts and liver tissue. A novel homozygous 22 base pair (bp) deletion in exon 1 of this enzyme's gene was identified (c.103_124del), which caused aframe-shift and subsequent premature stop codon. The “Wisdom pure breed-mixed breed” test reported the dog as a cross between a Miniature and Toy Poodle.

CONCLUSIONS: The clinicopathological features are similar to those of MPS type VI as previously described in dogs, cats and other species, and this clinical diagnosis was confirmed by enzymology and molecular genetic studies. This is an autosomal recessively inherited lysosomal storage disease.

CLINICAL RELEVANCE: The prevalence of MPS VI in Miniature or Toy Poodles in New Zealand and elsewhere is currently unknown. Due to the congenital nature of the disorder, malformed pups may be subject to euthanasia without investigation and the potential genetic problem in the breed may not be fully recognised. The establishment of a molecular genetic test now permits screening for this mutation as a basis to an informed breeding policy.     

A new gene <Emphasis Type="Italic">Bph33</Emphasis>(<Emphasis Type="Italic">t</Emphasis>) conferring resistance to brown planthopper (BPH), <Emphasis Type="Italic">Nilaparvata lugens</Emphasis> (Stål) in rice line RP2068-18-3-5

The rice brown planthopper (BPH) Nilaparvata lugens (Stål) is one of the major pests of rice across Asia. Host-plant resistance is the most ecologically acceptable means to manage this pest. A rice breeding line RP2068-18-3-5 (RP2068) derived from the land race Velluthacheera is reported to be resistant to BPH populations across India. We identified a new R gene [Bph33(t)] in this line using advanced generation RILs derived from TN1 × RP2068 cross through phenotyping at two locations and linkage analysis with 99 polymorphic SSR markers. QTL analysis through IciMapping identified at least two major QTL on chromosome 1 influencing seedling damage score in seed box screening, honey dew excretion by adults and nymphal survival. Since no BPH R gene has been reported on chromosome 1, we designate this locus as a new gene Bph33(t) which accounted for over 20% of phenotypic variance. Scanning the region for candidate gene suggested two likely candidates a leucine rich repeat (LRR) gene and a heat shock protein (HSP) coding gene. Expression profiling of the two genes in the two contrasting parents and RILs showed induction of the HSP gene (LOC_Os01g42190.1) at 6 h after infestation while LRR gene did not show such induction. It is likely that the HSP represented Bph33(t).