Evidence for classification of c.1852_1853AA > GC in MLH1 as a neutral variant for Lynch syndrome

Background: Lynch syndrome (LS) is an autosomal dominant inherited cancer syndrome characterized by early onset cancers of the colorectum, endometrium and other tumours. A significant proportion of DNA variants in LS patients are unclassified. Reports on the pathogenicity of the c.1852_1853AA>GC (p.Lys618Ala) variant of the MLH1 gene are conflicting. In this study, we provide new evidence indicating that this variant has no significant implications for LS. Methods: The following approach was used to assess the clinical significance of the p.Lys618Ala variant: frequency in a control population, case-control comparison, co-occurrence of the p.Lys618Ala variant with a pathogenic mutation, co-segregation with the disease and microsatellite instability in tumours from carriers of the variant. We genotyped p.Lys618Ala in 1034 individuals (373 sporadic colorectal cancer [CRC] patients, 250 index subjects from families suspected of having LS [revised Bethesda guidelines] and 411 controls). Three well-characterized LS families that fulfilled the Amsterdam II Criteria and consisted of members with the p.Lys618Ala variant were included to assess co-occurrence and co-segregation. A subset of colorectal tumour DNA samples from 17 patients carrying the p.Lys618Ala variant was screened for microsatellite instability using five mononucleotide markers. Results: Twenty-seven individuals were heterozygous for the p.Lys618Ala variant; nine had sporadic CRC (2.41%), seven were suspected of having hereditary CRC (2.8%) and 11 were controls (2.68%). There were no significant associations in the case-control and case-case studies. The p.Lys618Ala variant was co-existent with pathogenic mutations in two unrelated LS families. In one family, the allele distribution of the pathogenic and unclassified variant was in trans, in the other family the pathogenic variant was detected in the MSH6 gene and only the deleterious variant co-segregated with the disease in both families. Only two positive cases of microsatellite instability (2/17, 11.8%) were detected in tumours from p.Lys618Ala carriers, indicating that this variant does not play a role in functional inactivation of MLH1 in CRC patients. Conclusions: The p.Lys618Ala variant should be considered a neutral variant for LS. These findings have implications for the clinical management of CRC probands and their relatives.

White spot syndrome virus (WSSV) transmission risk through infected cooked shrimp products assessed by polymerase chain reaction (PCR) and bio-inoculation studies

The aim of the study was to evaluate the resistance of white spot syndrome virus (WSSV) in shrimps (Penaeus monodon) to the process of cooking. The cooking was carried out at 1000C six different durations 5, 10, 15, 20, 25 and 30 min. The presence of WSSV was tested by single step and nested polymerase chain reaction (PCR). In the single step PCR, the primers 1s5 & 1a16 and IK1 & IK2 were used. While in the nested PCR, primers IK1 &IK2 – IK3 & IK4 were used for the detection of WSSV. WSSV was detected in the single step PCR with the primers 1s5 and 1a16 and the nested PCR with the primers IK1 and IK2 – IK3 & IK4 from the cooked shrimp samples. The cooked shrimps, which gave positive results for WSSV by PCR, were further confirmed for the viability of WSSV by conducting the bio-inoculation studies. Mortality (100%) was observed within 123 h of intra-muscular post injection (P.I) into the live healthy WSSV-free shrimps (P. monodon). These results show that the WSSV survive the cooking process and even infected cooked shrimp products may pose a transmission risk for WSSV to the native shrimp farming systems.

Is epizootic ulcerative syndrome (EUS) specific fungus of fishes a primary pathogen?: an opinion

Earlier findings on epizootic ulcerative syndrome (EUS) and the present observation of the authors on transmission of EUS to snakehead (Channa sp.) without skin damage provide evidence to suggest that the invasive fungus associated with EUS is a primary pathogen.

Beverton and Holt equations: spreadsheet functions and uncertainty

This contribution illustrates how modern spreadsheets aid the calculation and visualization of yield models and how the effects of uncertainties may be incorporated using Monte Carlo simulation. It is argued that analogous approaches can be implemented for other assessment models of simple to medium complexity justifying wider use of spreadsheets in fisheries analysis and training.

Use of homeo drug for curing ulcerative syndrome in fishes

Fungal infection was observed in Catla catla and Labeo rohita cultured in two private fish farms. The later stage of the infection resulted in ulcerations followed by haemorrhage on the dorsal surface of the body. Initially, usual treatments of copper sulphate, potassium permanganate and common salt solution were tried, but no improvement was observed. Then repeated intramuscular injections of homeopathic drug Heaper Sulpher and Arnica spray were given with encouraging results. Infection reported in another farm was also successfully controlled using a similar treatment.

Incidence of epizootic ulcerative syndrome (EUS) in freshwater fishes in the endemic area of Punjab, Pakistan

Incidence of Epizootic Ulcerative Syndrome (EUS) has been recorded for the first time in freshwater fishes in the endemic area of Punjab, Pakistan. Survey of private fish farms, hatchery and natural water bodies was conducted in a radius of 14 Km from around river Ravi near Lahore (Punjab Province) Pakistan. Out Of 1628 fishes belonging to 18 genera, 517 fishes of 10 genera were found affected with EUS. The incidence of EUS in culturable fishes was higher in Cirrhina mrigala (15.4%) moderate in Catla cat/a (13.3%) and lower in Labeo rohita (5.0%). Exotic fish, Chinese carp Ctenoparyngodon idella and Hypophthalmicthys molitrix were not affected with EUS. In non-culturable fishes the incidence of EUS was highest in Channa punctatus (72.8%) moderate in by C. straitus (65.45%) and comparatively lower Puntius ticto (43.7%). A slow growing temperature sensitive Saprolegnia spp. was isolated from all of EUS infected fish species. Aeromonas spp. and Pseudomonas spp. were isolated from the diseased fishes. Ectoparasites viz. Lernaea, Argulus and Triclwdina spp. were also isolated from the skin and gills of infected fish species. The disease was more severe in water having low alkalinity (70 mg/1), hardness (75 mg/1) and low temperature of 10-12 °C.

Rapid detection of white spot syndrome virus (WSSV) of Penaeus monodon by latex agglutination test using monoclonal antibodies

Latex beads were sensitized with monoclonal antibodies (MAb) rose against VP28 of WSSV. The optimum concentration of MAb required to sensitize the latex beads was 125 µg/ml. The sensitized latex beads were used to detect WSSV from PCR-positive stomach tissue homogenates obtained from infected shrimp. Stomach tissue homogenates from WSSV-infected shrimp agglutinated the sensitized latex beads within 10 minutes, while uninfected samples did not produce any agglutination, although non-specific agglutinations were observed in some samples. The analytical sensitivity, analytical specificity, diagnostic sensitivity and diagnostic specificity of the (LAT) agglutination test were assessed. The analytical sensitivity of the test was 40 ng of purified WSSV (2 µg/ml). The sensitized latex beads did not agglutinate with normal shrimp tissue or MBV-infected tissue homogenate. The test has a diagnostic sensitivity of 70 and 45%, respectively, compared to single-step and nested PCR. The diagnostic specificity of the test was 82%. This test is a simple and rapid on-farm test which can be used to corroborate clinical signs for the detection of WSSV in grow-out ponds.

Polymorphisms in the C-type lectin genes cluster in chromosome 19 and predisposition to severe acute respiratory syndrome coronavirus (SARS-CoV) infection

Background: Polymorphisms of CLEC4M have been associated with predisposition for infection by the severe acute respiratory syndrome coronavirus (SARS-CoV). DC-SIGNR, a C-type lectin encoded by CLEC4M, is a receptor for the virus. A variable number tandem