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Subject: Veterinary Microbiology × End date: 2006 × Thesis Type: M.V.Sc. ×
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    ThesisItemOpen Access
    Role of parrots in the epizootiology of newcastle disease
    (Department of Microbiology, College of Veterinary and Animal Sciences,Mannuthy, 1981) Vijayan, V; Sulochana, S
    The incidence, susceptibility, mode of infection and duration of excretion of the new cattle disease virus in the common Indian parrots (psittacula Krameri) were studied in detail. The blood, cloacal and throat swabs of parrots, collected from different parts of the State were screened for the presence of ND antibodies and virus. Seventeen out of 103 blood samples were found to possess HI antibodies. The serum samples which gave positive HI titres belonged to parrots kept as pets in households and allowed to mingle freely with domestic poultry and those housed in Trivandrum Zoo along with pigeons. None of the 70 cloacal and 42 throat swabs were positive for the virus. Experimental infection of parrots with undiluted virulent virus by the intranasal and intraocular routes and by the subcutaneous and intranasal routes with 1:100 dilution of the same virus gave almost the same results. All of them died within a weeks’ time, after showing symptoms of inappettance, leg and wing paralysis and diarrhoea, from day two of infection. Virus could be isolated from the throat and cloacal swabs and also from the tissue of dead birds. Chicks kept along with these infected parrots did not develop symptoms of ND, eventhough they excreted the virus, for a few days, and had a low titre of HI antibodies in their sera. All the contact chicks died of ND with typical symptoms and lesions on challenge with the virulent virus. The parrots that received a mesogenic strain (Komorov) of the virus, also succumbed to the disease, but with less pronounced symptoms and with an extended incubation period. The parrots that were infected with lentogenic strain of the virus (F1) did not develop symptoms of the disease. However multiplication of the virus occurred in these birds as isolations could be made from cloacal and throat swab and a slight increase in H1 titre was noticed sera. However on challenge with a virulent strain of NDV, they showed symptoms of ND. All of them died within eight days and the virus could be isolated from them. Contact infection of parrots from infected chicks were quite effective, as the parrots died with the same symptoms described above, almost within the same time as direct infection. Virus was also isolated from the tissues of the dead parrots. The common Indian parrots were found to be highly susceptible to both velogenic and mesogenic strains of NDV, but they were resistant to the lentogenic strain. Uninfected chicks kept along with the parrots infected with virulent virus picked up the infection, and virus could be isolated from the cloacal and throat swabs of these chicks. They also showed an increase in the antibody titre. The failure to produce clinical disease in chicken might be attributed to a decrease in virulence of the virus on passage in parrots. The carrier state with the lentogenis strain of the virus could not be assessed as they were challenged after 2 weeks. Though a carrier role had been attributed to the parakeets imported from India, the parrots in this study were found to succumb to the disease within a week. This might be due to the variation in the susceptibility of various species of parrots to NDV. The chances of dissemination of the virus could be prevented by quarantining them for a period, not less than ten days.
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    ThesisItemOpen Access
    Isolation and identification of viruses from waterfowls seen in Kerala
    (Department of microbiology, College of veterinary and animal sciences, Mannuthy, 1997) Bindu, M S; KAU; Krishnan Nair, G
    In the recent years, frequent outbreaks of poultry diseases have caused significant economic losses and among the various sources of infection, freeflying migratory birds and waterfowls are reported to be an important source for the introduction of disease to domestic flocks. These freeflying pirds and waterfowls abound in the waterlogged areas of Kerala especially Alleppey, Kottayam, Thrissur and Malappuram districts. Heavy losses were reported each year due to severe outbreaks of duck plague and duck pasturellosis resulting in the wiping out of the duck population in the state. This may be due to the introduction of the disease agents by migratory waterfowls. Hence a study was undertaken to elucidate the role of waterfowls in the spread of diseases to domestic poultry and ducks. A total of 52 waterfowls were caught from different parts of Kerala of which 15 were lesser whistling teals and 37 were gargany. Postmortem examination of these birds were carried out and the required materials were collected. Impression smears from the cut surface of the liver revealed intranuclear inclusion bodies in the hepatic cells in four cases (Bird Nos. 17, 23, 27 and 49) and two haemagglutinating agents were isolated from the cloacal swabs of the bird numbers 18 and 22. The haemagglutinating agents developed lesions in the chicken embryos which died 3 to 5 days after inoculation. the allantoamniotic fluid of the affected embryos agglutinated teal and chicken erythrocytes. The lesions exhibited by the embryos infected with Ta were congestion of the CAM and embryos while in T22 the embryos were stunted, curled and at the same time congested. The liver of the embryo had yellowish brown patches. The haemagglutination activity of both the viral isolates were tested with red cells of various species like cattle, horse, human '0', duck, rabbit and pig but was negative in all cases. Both the isolates lost their infectivity and haemagglutination property at 56°C for 30 min. the infectivity and HA activity of the viral isolates T18 and T22 were retained at pH 7.2 and were completely destroyed at pH 3.2 and pH 9. Both the isolates were sensitive to treatment with chloroform, revealing both as enveloped viruses, wherein the infectivity was completely lost and HA activity was considerably reduced. In the case of Tl8 HA activity was reduced from 512 to 16 and for T22 from 128 to 8. The nucleic acid types of the viral isolates were confirmed by inoculating the isolates to chicken embryo fibroblast cultures pretreated with 100 µg and 200 µg per ml of IudR which led to the conclusion that Ta was a RNA virus and T22 a DNA virus. The ELD50 of Ta and T22 were 107 ELD50/ml and 106 ELD50/ml. The ICPI, MDT and IVPI. were calculated as for NDV. The ICPI for the isolates were 0.83 and 0.30 for T18 and T22 respectively, MDT was 120 hr for both and the IVPI was calculated to be zero in both cases, indicating that both the isolates were nonpathogenic. In cell cultures both the isolates produced CPE which affected the whole monolayer by 96 hrs. In the case of Tl8 the CPE was characterised by rounding and clumping of cells, the infected cells showing a tendency to get separated from the neighbouring cells leaving long cytoplasmic strands, syncytium formation with four or five nuclei and severe cytoplasmic vacuolation. For T22 the CPE was characterised by rounding and cytoplasmic vacuolation and karryorhexis. Intranuclear and intracytoplasmic inclusion bodies could be demonstrated. The pathogenicity studies of both the isolates were carried out in one week old and six week old ducklings and chicken, for which both were nonpathogenic and did not exhibit any clinical signs or mortality. But viral infection was established in six week old chicks by virus isolation till the 14th day of infection from the cloacal swabs for both the isolates. The sera collected from these birds revealed an antibody titre of 16 for T¬18 and 8 for T22 indicating the infection. The antigenic relationship of the isolates was examined with NDV, EDS-76 and fish viruses (FV and F6), of which T22 did not show any antigenic similarity with any of the viruses. But T1B on the contrary exhibited antigenic relationship with the fishviruses FV and F6 but no antigenic similarity with NDV and EDS-76. The antigenic similarity exhibited by T18 with the fish viruses leads to the conclusion that waterfowls may be disseminating the viruses responsible for the outbreak of epizootic ulcerative syndrome in fishes. The morphological features of T18 by electronmicroscopy revealed an enveloped virus with a size of 150-185 nm with pleomorphic forms and peplomers of a length of 18-20 nm. Except the length of the peplomers it similated a paramyxovirus. The morphology of T22 was not studied due to technical defects.
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    ThesisItemOpen Access
    Antigens of pasteurella multocida isolates from rabbit and their immunologencity
    (Department of microbiology, College of veterinary and animal sciences, Mannuthy, 1994) Manoharan S; KAU; Jayaprakasan, V
    Two rabbit strains viz. R9 S and R23 S and a bovine vaccine strain P – 52 which were maintained in virulent form, were used for the preparation of three forms of antigen viz., heat inactivated crude extract, KSCN extract and sonicated antigen. These antigens were chemically analysed for protein and carbohydrate contents and were found to be higher in the sonicated antigen preparation irrespective of the source. In SDS – PAGE analysis, the profiles discerned by heat inactivated crude extract, KSCN extract and sonicated antigens were four, five and six protein bands with molecular weights lesser than 68 kDa while the KSCN extract and sonicated antigen presented an additional protein band with molecular weight higher than 68 kDa. Three types of antigen of P. multocida were characterized and analysed for the inter relationship and the immunogenic potential in mice. Antiserum was raised against each antigenic preparation from the three strains in rabbits and used for serological study. In AGPT and immunoelectrophoresis the serum developed multiple precipitin lines and arcs respectively when reacted against the three homologous and two heterologous antigens in which a few were identical to the heterologous antigens. The results revealed stronger serological relationship between the two rabbit strains than with the cattle strain and the heterogeneity of the sonicated antigen. The antibody titre in each antiserum was measured by IHA using the sensitized GA – SRBC/T – GA – SRBC and the titres were more in the homologous antiserum and high titre for the heterologous serum was seen with the sonicated antigen. The LD50 determined for the three strains R9 S, R23 S and P- 52 was found to be 3 x 104 , 3 x 103 and 3 x 105 bacteria. Immunogenic potential of the three antigens and an adjuvanted sonicated antigen were tested in mice by giving two doses of vaccine at 14 days interval and challenging on 21st day with homologous and heterologous strains. A higher percentage of protection was conferred by homologous strains and it was cent per cent (100%) with sonicated antigen. The percentage of protection against challenge with heterologous strains was low. An elaborated study on immunity trials with these immunogens is needed before recommending the R23 S as a candidate vaccine strain.
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    ThesisItemOpen Access
    Characterization of staphylococci isolated from cases of mastitis and study of their R plasmids
    (Department of microbiology, College of veterinary and animal sciences, Mannuthy, 1986) Mathew, E S; KAU; Punnoose, K T
    The emergence of drug resistant staphylococci causing mastitis deserve serious investigation. The work was intended to characterize staphylococci isolated from cases of bovine mastitis and to study their R plasmid transfer in vitro. The characterization was done using glucose and mannitol fermentation, catalase production, coagulase production, Iysostaphin sensitivity and bacteriophage typing. The antibiogram of the isolates was done by agar diffusion method using 14 chemotherapeutic agents (amoxicillin, ampicillin, bacitracin, chloramphenicol, cloxacillin, erythromycin, gentamicin, methicillin, neomycin, nitrofurantoin, penicillin, streptomycin, sulphamethoxazole and tetracycline) and by agar dilution method using eight antibiotics (ampicillin, chloramphenicol, rifampicin, streptomycin, tetracycline , gentamicin, erythromycin and penicillin) The in vitro transfer of R plasmids was tried using selected S.aureus and coagulase-negative staphylococcal isolates as donors and S.aureus RN 450RF and S.epifermidis 131S as recipients. From 360 milk samples collected from cases of bovine mastitis 17 strains of S.aureus and 35 strains of coagulase negative staphylococci were isolated. Lysostaphin sensitivity test was positive for 90.38% of the isolates, but this could not be used to differentiate between coagulase positive and negative staphylococci. The results of phage typing revealed a predominance of group III phages over the other groups and the possible role of human strains of staphylococci in producing mastitis in animals. None of the strains were resistant to mercuric chloride. From the antibiogram rifampicin , bacitracin, neomycin , methicillin, gentamicin, cloxacillin, nitrofurantoin and chloramphenicol were found to be the drugs of choice in the treatment of bovine mastitis caused by staphylococci. When S. aureus Rn 450RF was used as recipient six of the ten selected S.aureus isolates could transfer either one or more drug resistance markers and the mode of transfer was suspected to be by conjugation. All the four selected streptomycin resistant coagulase –negative staphylococcal donors were found to transfer the R plasmid DNA to the recipient., S.aureus RN 450RF, which was established to be through conjugation. The drug resistant S. aureus as well as coagulase negative staphylococcal strains failed t transfer their resistance to S.epidermidis 131S.
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    ThesisItemOpen Access
    Characterization and immunology of inflenza virus type a isolated from duck in Kerala
    (Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 1988) Mini, M; KAU; Sulochana, S
    Ducks are generally regarded as resistant to influenza virus infection. During the early halves of 1985 and 1987 influenza A viruses were isolated from cases of respiratory disease causing 15 to 20 percent mortality in two to six week-old ducklings at Government Duck Farm, Niranam. Four strains of the virus (A/duck/India /1/85 (H9N2)-CDN, A/duck/India /2/87 (H3N?) –DT3; A/duck/India /7/87 (H3N2)-T19 and A/duck/India /14/87 (H9N?-lgl isolated from these out breaks were studied in detail with particular reference to their characteristics, Pathogenicity and immunogenicity. The strains were propagated in nine day embryonated chicken eggs by allantoic route of inoculation. All the four strains multiplied well killing the embryos in two to four days time. Though characteristic lesions were not present, the embryos were slightly congested. The chorioallantoic membranes showed moderate congestion and oedema but no pock lesions. Infectivity titers were 108.5 EID 50/0.2ml; 108.25 EID 50/0.2ml 105.75 EID 50/0.2ml and 108.5 EID 50/0.2ml respectively for CDN, DT3, T19 and lgl. The corresponding HA titres were 1:64 , 1:128, 1:64 and 1:256 The strains also multiplied well in duck embryos but no specific lesions were seen either in the embryo or on the chorioallantoic membranes. The infectivity titers were low compared to the titers obtained in chick embryos. All the strains were inactivated at 560C in 30 minutes. They lost their infectivity at PH 3.2 while HA property was not considerably reduced except for lgl. At PH 7.2 both infectivity and HA property were not affected. But infectivity was slightly affected and HA property was markedly reduced at PH 9.0. The viruses were chloroform sensitive and agglutinated red cells from cattle, sheep, goat, guinea pigs, horse, rabbit, rat mouse, chicken and man (0, A and B Groups). Chicken embryo fibroblastic infected with strain CDN produced cytopathic effects characterized by rounding of cells affecting the whole monolayer in 96 hours. The remaining three strains did not produce any CPE. In all the four cases virus infection was evidenced as the cell culture fluid gave haemagglutination. The infectivity titers of these fluids were 106.5 EID 50/0.2ml and 105.5 EID 50/0.2 ml 104.0 EID 50/0.2 ml and 105.5 EID 50/0.2 ml respectively for CDN, DT3, T19 and lgl. Two common antigens possibly the type specific MP and NP antigens were detected in the CAM extracts of embryos infected with these strains by agar gel precipitation and immunoelectrophoresis. Mean death time for these strains calculated according to the method adopted for NDV were 78 hours, 76.8 hours, 72 hours and 76 hours respectively for CDN, DT3, T19 and lgl, while the ICPI in day-old chicks were 0.325, 0.66, 0.00 and 0.16 and IVPI 0.00 in all cases. Pathogenicity and immunogenicity studies were carried out in day-old, one week-old and two week-old ducklings with strain CDN and in the week-old ducklings with strain lgl. Strain CDN produced hundred percent mortality in day old ducklings that received the virus by oral , occulonasal and subcutaneous routes. They died with no marked clinical signs except droopiness, discharges from the eyes. Slight diarhoea and ruffled feathers in some of them. The pathogenicity indices were 1.19, 1.15 and 1.51 respectively for group A,B and C. The ducklings that were kept to study contact infection revealed cloacal excretion of the virus till 12th day, through tracheal excretion stopped by 10th day. None of the birds in all the four groups that survived seven day of infection did reveal any specific seven day of infection did reveal any specific HI antibodies. In one week-old ducklings no clinical symptoms were observed. Virus could be isolated from cloacal and throat swabs before death and from tissues of dead birds. The pathogenicity indices were 0.36, 0.00 and 0.20 respectively for group A, B and C. In contact ducklings picked up infection as indicated from virus isolation and antibody titration. Both infected and incontact birds showed HI antibodies from 7th day onwards. By 14th day the titres reached peak level ranging between of 1:16 to 1:64 followed by a decline and remained steady through out the observation period. The two week-old ducklings did not show any clinical symptoms or death. But they revealed tracheal and cloacal shedding of the virus. The HI antibody titres never increased beyond 1:16. None of the incontent birds either showed clinical symptoms or death. Positive virus isolation could be made till the 7th day. Antibody response was also very poor and HI antibody titers never increased beyond 1:4. One week-old ducklings infected with strain lgl remained apparently healthy throughout the period of observation. However, virus isolations were made from cloacal and tracheal samples from 3rd to 7th day. In birds that received the virus by oral route, HI antibody titres were 1:4, 1:8 and 1:4 respectively on 7th, 14th and 21st day and nil by 28th day. In order two groups the birds did not reveal HI antibodies till 28th day. None of the incontact birds in all three groups showed clinical symptoms, death or any other indication of infection by the virus. In addition the sera from these birds were also negative for HI antibodies.
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    ThesisItemOpen Access
    Secretory immunoglobulins of the duck (Anas platyrrhynchos domesticus)
    (Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 1996) Hari, Kumar A V; James, P C
    The profile and functional properties of the immunoglobulins of serum, bile, mucos of trachea and intestine of ducks were studied. The immunoglobulins were separated by salting out using ammonium sulphate. The various fractions of immunoglobulines were further resolved by Sephadex G-200 gel filteration which gave two peaks for serum and tracheal immunoglobulins and a single peak each for bile and intestinal immunoglobulins. The purity of these fractions were checked by immunodiffusion and immunoelectrophoresis. The different fractions obtained were quantified by single radial immunodiffusion. The level of fraction 1 in the bile ranged between 1718 µg/ml and 1959 µg/ml and that of serum, 1718.06 µg/ml to 2442 µg/ml, in the S.typhimurium treated groups. The level of fraction 1 in the NDV treated groups ranged from 1115 µg/ml to 1597.35 µg/ml in bile, and 1597.35 µg/ml to 1959.34 µg/ml in serum. The level of fraction 2 in serum ranged from 1797 µg/ml to 2591 µg/ml in S.typhimurium treated group and 1400 µg/ml to 1797 µg/ml in the NDV treated group. Fraction 2 was not detectable in bile. The antibody response of ducks to a bacterial and viral antigen (anaculture of S.typhimurium and R2B strain of New Castle Disease virus respectively) was assessed. On conducting standard tube agglutination test, the serum, bile and oviduct washings revealed antibody tires against S.typhimurium in inoculated birds ranging between 1:20 and 1:160 in the case of serum; 1:10 and 1:80 in the case of bile and tire less than 1:10 for oviduct washings. No antibody titre could be detected for tracheal and intestinal washings and testicular extracts. The HI titre ranging from 1:32 to 1:128 could be observed for serum, 1:32 to 1:64 for bile and a titre of 1:16 was observed for oviduct washings of ducks parenterally administered with NDV. The tire was relatively low for serum when NDV was administered intranasaly. Intestinal and tracheal washings and testicular extract failed to reveal any HI antibodies.
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    ThesisItemOpen Access
    Survey on the incidence of Salmonellae in meat animals
    (Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 1977) James, P C; Abdulla, P K
    Prior to this “Survey on the Incidence of Salmonellae in Meat Animals” the information on the serotypes of Salmonella prevalent and the magnitude of their occurrence in livestock in Kerala had been meagre except for the reports by Sulochana et al. (1973) and Balakrishna Pillai (1975). The work carried out during the present investigation has gathered more information on the prevalence of Salmonella in livestock in Kerala. In this study the prevalence of Salmonella serotypes in the different species of animals was probed. A total of 823 biomaterials, besides 50 drain samples were subjected to cultural screening. This venture resulted in the recovery of 56 strains of Salmonella. Serological identification of many of these strains proved the prevalence of S. typhimurium, S. weltevreden and S. urbana. The preponderatingly prevalent serotypes were found to be the former two. Pathogenicity studies employing S. typhimurium culture in mice, guinea pigs, rabbits, calves and piglets were conducted. All these animals were found to be susceptible to infection by S. typhimurium evincing varying degrees of clinical manifestations. The advantages of employing modified McConkey’s medium (Sharma, 1961) containing Mannitol instead of lactose and composite medium 1 and 11 developed by Chitin is et al. (1972) to differentiate Salmonella at primary screening level have been discussed
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    ThesisItemOpen Access
    Studies on the bacterial species associated with gastroenteritis in goats
    (Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 1979) Sebastian, Joseph; KAU; Abdulla, P K
    The information regarding the incidence, etiology and pathogenicity of enteric pathogens in goats is very meagre in our country. The present study is aimed at the isolation, identification and characterisation of Enterobacterial organisms from cases of enteritis in goats. The study also included, determination of sensitivity pattern of the isolates to various chemotherapeutic agents. A total of 190 specimens, which included rectal swabs (60), intestinal contents, portions of large and small intestines (92) and mesenteric lymph nodes (38) collected from live/dead animals were examined for enteric pathogens. From these specimens examined, 86 isolates of Escherichia coli (45.26 per cent), 39 Enterobacter cloacae (20.33 per cent) and two Salmonella (1.05 per cent) were obtained. Of all the E.coli isolates, only one (EC/11) was found to be haemolytic. In addition to the above specimens, eight samples of heart blood and 34 specimens of lung tissues collected from cases of gastroenteritis were also examined for the presence of bacterial organisms. Seven isolates of Streptococcus pyogenes (from lung tissues only), 15 isolates of Klebsiella Pneumoniae (from lung tissues only), and one isolate of Corynebacterium pyogenes (from lung tissues only) were obtained. The ability of haemolytic E.coli (EC/11) to produce necrotoxin on rabbit skin was tested and the lesions produced were of necrotic changes. The strain was also found to be pathogenic to mice when tested. One isolate of Salmonella (S/1) was also tested for its pathogenicity to mice, and found non – pathogenic. Enterotoxin production in rabbit ileal loop was studied with haemolytic (EC/11) and non – haemolytic (EC/15) E.coli. The test materials included peptone water culture, soft agar culture fluid and acetone precipitated culture fluid. The results of the experiment have shown that, non – haemolytic E.coli produced dilatation reaction, while the haemolytic E.coli did not. The lesions noticed in the ileal segments of positive reaction were typical of enteritis. Antibiotic sensitivity studies were conducted using 11 chemotherapeutic agents (Ampicillin, bacitracin, chloramphenicol, erythromycin, gentamicin, kanamycin, nitrofuran, penicillin, streptomycin, sulphonamide and tetracycline) on E.coli Salmonella and Enterobacter cloacae. The result showed that cent per cent isolates of E.coli were sensitive to gentamicin, 95.35 per cent to nitrofuran, 88.37 per cent to chloramphenicol, 60.47 per cent to kanamycin, 40.70 per cent to streptomycin, 8.14 per cent to tetracycline and 2.33 per cent to erythromycin. All the 39 isolates of Enterobacter closcae tested were sensitive to gentamicin and kanamycin, whereas 30 (76.92 per cent) were sensitive to chloramphenicol and nitrofuran and 15 (38.46 per cent) to streptomycin. The drugs of choice for Salmonella were found to be gentamicin, chloramphenicol, nitrofuran and streptomycin.
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    ThesisItemOpen Access
    Immunology survey on the incidence of infectious bronchitis(IB) and infectious laryngotracheitis (ILT) in poultry in and around Trichur
    (Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 1979) George, M C; KAU; Punnoose, K T
    Infectious bronchitis and infectious laryngotrancheitis are the two viral diseases of poultry responsible for economic loss to the poultry industry by way of decreased egg production, poor quality of eggs, decreased feed efficiency and loss of weight gain. These disease have been reported from the neighbouring states of Kerala. In the present study a serological survey was carried out to understand the prevalence of these two disease in the poultry population in and around Trichur. A total of 2,110 serum samples have been collected from the field, comprising of white leghorn, Rhode Island Red and Desi birds belonging to different age groups. Serum samples were collected from organized farms, from birds kept by farmers and from the birds slaughtered in different hotels at Trichur. These serum samples were tested against the infectious bronchitis and infectious laryngotracheitis by employing agar gel precipitation test. The chorioallantoic membrane and allantoic fluid of infected embryos were used for the preparation of antigens for agar gel precipitation test. The potency of antigens was tested by conducting the agar gel precipitation test with corresponding hyper immuns sera prepared in white leghon male chicks of six to eight weeks of age. A line of precipitation was obtained in both cases which was close and curved towards the antigen well, because of the high concentration of antibody in the sera and due to the high molecular weight of the antigen. In the case of infectious bronchitis the line of precipitation was distinct where as in case of infectious laryngotrachetis it was diffused. The antigen, whose efficiency was tested using hyper immune sera, was used to test samples of sera collected from the field. The samples were pooled to 211 groups and tested for the presence of infectious bronchitis and infectious larygotracheitis precipitating antibodies separately by agar gel precipitation test. None of the samples gave precipitin line either to infectious bronchitis or to infectious larygotracheitis. So it was assumed that both of these viral disease are not prevalent in Trichur and its suburbs.