Infectious dose-dependent accumulation of live highly pathogenic avian influenza H5N1 virus in chicken skeletal muscle-implications for public health.

Highly pathogenic avian influenza viruses (HPAIV) of H5N1 subtype are a major global threat to poultry and public health. Export of poultry products, such as chicken and duck meat, is a known source for the cross-boundary spread of HPAI H5N1 viruses. Humans get infected with HPAI H5N1 viruses either by close contact with infected poultry or through consumption of fresh/undercooked poultry meat. Skeletal muscle is the largest soft tissue in chicken that has been shown to contain virus during systemic HPAIV infection and supports productive virus infection. However, the time between infection of a chicken with H5N1 virus and presence of virus in muscle tissue is not yet known. Further, it is also not clear whether chicken infected with low doses of H5N1 virus that cause non-fatal subclinical infections continue to accumulate virus in skeletal muscle. We investigated the amount and duration of virus detection in skeletal muscle of chicken experimentally infected with different doses (102 , 103 and 104 EID50 ) of a HPAI H5N1 virus. Influenza viral antigen could be detected as early as 6 hr after infection and live virus was recovered from 48 hr after infection. Notably, chicken infected with lower levels of HPAI H5N1 virus (i.e., 102 EID50 ) did not die acutely, but continued to accumulate high levels of H5N1 virus in skeletal muscle until 6 days post-infection. Our data suggest that there is a potential risk of human exposure to H5N1 virus through meat from clinically healthy chicken infected with a low dose of virus. Our results highlight the need to implement rigorous monitoring systems to screen poultry meat from H5N1 endemic countries to limit the global spread of H5N1 viruses.

Pathology of a H5N1, highly pathogenic avian influenza virus, in two Indian native chicken breeds and a synthetic broiler line.

In this study, susceptibility to H5N1 virus infection was studied in two Indian native chicken breeds viz. Kadaknath and Aseel (Peela) and an Indian synthetic broiler strain (Synthetic dam line (SDL-IC). Fifty birds from each genetic group were infected intra-nasally with 1000 EID50 of a highly pathogenic avian influenza virus (HPAIV) strain A/chicken/Navapur/India/7972/ 06 (H5N1) and observed for a period of 10 days. Significant differences in severity of clinical signs, gross lesions and time for onset of symptoms were observed. The overall severity of clinical signs and gross lesions was less in SDL-IC broilers as compared to the other two genetic groups. The mortality percentages were 100, 98 and 92% with Mean Death Time (MDT) of 3.12, 5.92 and 6.96 days, respectively for the two native breeds Kadaknath and Aseel (Peela), the and SDL-IC broiler strain. Comparison of histological lesions revealed differences in disease progression among the genetic groups. Vascular lesions such as disseminated intravascular coagulopathy (DIC) were predominant on 3 days post infection (dpi) in Kadaknath, and on 5 and 6 dpi in Aseel (Peela) and SDL-IC broiler. The mean log2 HA titres of the re-isolated virus from various organs of H5N1 AIV infected birds of the three genetic groups ranged from 2.32 (lung, trachea and bursa) to 5.04 (spleen) in Kadaknath; 2.32 (lung) to 6.68 (brain) in Aseel (Peela); and 2.06 (liver) to 7.01 (lungs and kidney) in SDL-IC broiler. These results suggest that the susceptibility to H5N1 highly pathogenic avian influenza virus infection differed among the three breeds; Kadaknath being highest followed by Aseel (Peela) and synthetic SDL-IC broiler. This is possibly the first report on the differences in the susceptibility of the India native breeds to H5N1 virus infection and its severity.

Influence of dose of inocula on outcome of clinical disease in highly pathogenic avian influenza (H5N1) infections--an experimental study.

Twelve-week-old Vanaraja (an Indian native dual purpose breed) chickens were inoculated intranasally with different doses (100, 1000, and 10,000 mean embryo infective dose [EID50]) of H5N1 virus, and the clinical disease and pathologic changes were compared. Although the overall severity of clinical signs was more severe in the 100 EID50 group, the progression of the clinical disease was slower with delayed onset of mortality when compared with the other two groups. The mean death time of the 100 EID50 group (4.57 days) differed significantly from that of the 10,000 EID50 group (3.60 days) and from that of the 1000 EID50 group (3.33 days). Similarly, overall severity of gross lesions was expressed more in the 100 EID50 group. The histopathologic lesions were of a more hemorrhagic and necrotic nature in the 100 EID50 group, histopathologic lesions were of an inflammatory/proliferative nature in the 1000 EID50 group, and a tendency for intravascular coagulopathy was observed in the 10,000 EID50 group. These differences may be assigned to the influence of dose in the outcome of disease.

Genotoxicity of a polluted river system measured using the alkaline comet assay on fish and earthworm tissues.

Monitoring genotoxicity in the environment by using endemic organisms as sentinels requires sensitive assays. In this study the genotoxic properties of water and sediment collected from the Noyyal River, which is polluted with industrial effluent and sewage, was determined in fish (Cyprinus carpio) and earthworms (Eisenia foetida) using the alkaline comet assay. Upon electrophoresis, extensive DNA damage, measured as the DNA length:width ratio of the DNA mass, was observed in erythrocytes, liver, and kidney cells of fish exposed to polluted water samples and the amount of damage increased with the duration of exposure. Similarly, the mean DNA length:width ratio was significantly higher in the coelomocytes of earthworms placed in sediment samples. The highest levels of DNA damage were obtained with samples taken at and immediately downstream of urban centers. The results of this study indicate that the Noyyal River system is contaminated with substances that are genotoxic to fish and earthworms and that the comet assay has sufficient sensitivity to detect the genotoxicity.

Uptake of dyes by a promising locally available agricultural solid waste: coir pith.

The adsorption of rhodamine-B and acid violet by coir pith carbon was carried out by varying the parameters such as agitation time, dye concentration, adsorbent dose and pH. The adsorption followed both Langmuir and Freundlich isotherms. The adsorption capacity was found to be 2.56 mg and 8.06 mg dye per g of the adsorbent for rhodamine-B and acid violet, respectively. Adsorption of dyes followed first order rate kinetics. Acidic pH was favorable for the adsorption of acid violet and alkaline pH was favorable to rhodamine-B. Desorption studies showed that alkaline pH was favorable for the desorption of acid violet and acidic pH was favorable for the desorption of rhodamine-B.

Genotoxicity of some sulfur dyes on tadpoles (Rana hexadactyla) measured using the comet assay.

This report presents the results of a genotoxicity study to evaluate the DNA damage caused by four sulfur dyes used in the textile and tannery industries. Alkaline single-cell gel electrophoresis assay (SCGE) was performed on erythrocytes from Rana hexadactyla tadpoles following whole-body exposure to increasing concentrations of the dyes. The dyes, along with their active ingredients, were Sandopel Basic Black BHLN, Negrosine, Dermapel Black FNI, and Turquoise Blue. The dye-treated tadpoles showed significant DNA damage, measured as mean DNA length:width ratio, when compared with unexposed control animals. Among the four tested dyes Sandopel Basic Black BHLN appears to be highly genotoxic, Dermapel Black FNI was least genotoxic, and Negrosine and Turquoise Blue were moderately toxic to R. hexadactyla tadpoles. The tadpoles showed a significant reduction in DNA damage when placed in dechlorinated tap water after exposure for a 24-hr period to the dye solutions.

Quantitative assessment of circulating antigens in human lymphatic filariasis: a field evaluation of monoclonal antibody-based ELISA using blood collected on filter strips.

OBJECTIVE: To quantify circulating antigens in individuals with lymphatic filariasis by means of an ELISA using blood on filter strips. METHOD: Circulating antigens in filarial patients and normal individuals living in an area endemic for W. bancrofti infection in Madras, India were estimated using a monoclonal antibody-based ELISA. RESULTS: All microfilaraemics showed positivity to circulating antigens whereas people with chronic pathology and 80% of the endemic normals tested negative. The antigen levels in the blood collected in the night and during day time showed positivity and there was no difference in the antigen concentration. The results of the antigen levels collected onto filter strips correlated with their corresponding plasma antigen levels (r = 0.83). In microfilaraemics, DEC treatment did not alter the levels of circulating antigens for up to one month. CONCLUSIONS: We conclude that this monoclonal antibody-based ELISA using filter strips may be used in day time and replace the existing routine night blood surveys in our endemic area in India.

Venous allografts for infrainguinal vascular bypass.

Between 1978 and 1993, 107 patients received venous allografts for infrainguinal revascularization because autogenous material was unavailable. These operations comprised 7.4% of 1442 reversed vein bypass procedures performed during the same period. Alloveins were harvested during varicose vein stripping and stored up to 10 days in saline solution containing heparin and antibiotics. Veins with or without minimal degenerative changes were used and when necessary segments were sutured end-to-end to obtain a convenient length. The patients comprised 95 men and 12 women of age range 44-87 years. All operations were performed for limb salvage (rest pain and gangrene); 40 femoropopliteal and 67 femorocrural bypasses were performed; 66 were primary reconstructions and 41 secondary reconstructions. The operative mortality rate was 3.7%. Early thrombosis occurred in 18 patients (16.8%), nine of whom were successfully reoperated upon. The cumulative patency rates for all bypass operations was 82.9%, 38.7% and 21.3% at 30 days, 3 and 5 years, respectively, the limb salvage rates being 84.8%, 48.0% and 29.2%, respectively. Allografts may have a place in lower-limb bypass surgery in the absence of veins, polytetrafluoroethylene or human umbilical vein.

Severe hypernatremia in a patient with psychiatric illness.

A 17-year-old schizophrenic developed severe hypernatremia during a period of psychosis. The thirst-deficient abnormality that caused this hypernatremia resolved when his psychosis improved. The primary disorders causing a thirst deficiency leading to hypernatremia fall into three categories: lesions of the central nervous system, mineralocorticoid excess, and drug side effects. None of these disorders was found in our patient. We conclude that psychosis can severely impair the thirst mechanism directly.