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Open 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.
Open Access
Cellular and humoran immune responses to corynebacterium presudotuberculosis infection in goats
(Department of microbiology, College of veterinary and animal sciences, Mannuthy, 1986) Jayaprakasan, V; KAU; Sulochana, S
Caseous lymphadenitis was experimentally produced in cross-bred malabari goats aged 8 to 12 months by bilateral inoculation of 1 x 106 viable C. pseudotuberculosis (ATCC 19410) through intradermal, subcutaneous and submucosal routes. The clinical picture, immune response and pathological features were studied up to a period of 13 weeks. The development of immune response in experimentally infected goats was assessed, by comparing the data with those of the controls, with respect to total serum protein, serum protein fractions, antibody activity of the serum, leukocyte count, counts of lymphocyte subpopulations, leukocyte migration inhibition index and skin hypersensitivity reaction. Pathological features in the lymphnodes and other tissues of infected goats necropsied at 15 days interval were also studied. Initial febrile reaction which lasted for 72 to 96 h, local inflammatory changes caused at the site of inoculations during the first two to three weeks of infection and the development of lesions typical of caseous lymphadenitis in local/regional lymphnodes within 21 days post-inoculation were the main features of clinical manifestations of the disease. aS a result of infection, neutrophilic leukocytosis was maximum during the 2nd week of infection. No appreciable was maximum during the 2nd week of infection. No appreciable change in counts of other cells in terms of their absolute numbers was noted during the entire period of observation. The humoral immune response in infected goats was indicated by the rise in serum protein, antitoxic antibody and B-lymphocyte count in the peripheral blood. The serum protein concentration increased to significant levels from the 5th week onwards and it reached the peak value (11.346 g%) by the 8th week. From the 3rd week onwards haemolysis inhibition test, which detected goats and persisted till the end of the observation period. The peak antibody level was recorded by the 5th week of infection and thereafter there was gradual reduction in the titre. Significantly high percentage of B- lymphocyte was recorded in infected animals from the 2nd to 10th week, except at the 5th week. The percentage of B-cells in infected goats ranged between 12.3 + 0.85 – 17.63 + 1.2 while it was 8.56 + 0.75-12.3 + 1.09 in control goats. This was considered as an indication of stimulation of humoral immune response. The cell-mediated immune response was evidenced by the increased T- lymphocyte count in the peripheral blood, inhibition of leukocyte migration and the development of delayed skin hypersensitivity. The mean percentage of T-lymphocytes in the peripheral blood of infected goats by E-rosette assay recorded an initial reduction at the first week (18.44 + 1.4) followed by an increase which was significant during the 5th, 6th, 7th, 8th, 12th and 13th week of infection. The maximum value was recorded (35.24 + 1.58) at the 13th week. In the case of control goats the percentage values ranged from 24.55 + 3.66 to 26.74 + 1.34. The T-lymphocyte count in the peripheral blood enumerated the ANAE method did not show any significant change even after infection. In the experimentally infected goats, leukocyte migration inhibition index was less than 0.8 during post-infection period while the control goats had the index value above 0.8. Significant reduction in the migration index was noted by 45th day of infection and the maximum reduction was on the 60th day. Intradermal injection of the toxic supernatant of the culture elicited characteristic delayed skin hypersensitivity reaction in all the experimentally infected goats while there was no reaction in the controls. The positive reaction was found to be maximum by the 48th hour post-injection. The pathological changes were characterized by an initial stimulatory hyperplastic reaction in the lymphnodes and this was accompanied by necrobiotic changes typical of caseous lymphadenitis. The hyperplastic stimulatory reactions were characterized by the presence of several active follicles with well developed germinal centres in the cortex, distinct medullary cords densely lined with plasma cells and sinus histiocytosis indicating the early elicitation of humoral immune response to the bacterium or to its in vivo products. The results obtained from the present study revealed the operation of both cell-mediated and humoral immune responses in goats against C. pseudotuberculosis infection. Of the various methods employed to monitor the immune response, leukocyte migration inhibition and delayed skin hypersensitivity tests were found to be of value in assessing the cell-mediated immune response and haemolysis inhibition test for humoral immune response. Leukocyte migration inhibition test and haemolysis inhibition test could be employed for early diagnosis of C. pseudotuberculosis infection in goats. FINDINGS : The immune responses and pathological features in Corynebacterium pseudotuberculosis infection were studied by experimental infection of cross- bred Malabari goats of S-12 months of age. Single cell bacterial suspension in chilled sodium chloride bile salt solution was used for this purpose. Goats were inoculated at both sides of the body by three routes viz., intradermal, subcutaneous and submucosal, with 2 x 106 bacteria per site of injection. The experimentally infected and control goats were observed for clinical manifestations of caseous lymphadenitis for a period of 13 weeks. The development of immune response in experimentally infected goats was assessed by comparing the data with those of the controls with respect to total serum protein, serum protein fractions, antibody activity of the serum, leukocyte counts, counts of lymphocyte sub-populations, leukocyte migration inhibition index and skin hypersensitivity reaction. Gross and histopathological changes in the lymphnodes and other tissues of necropsied goats were studied at 15 days interval for a period of 90 days. All experimentally infected goats exhibited rise in temperature, general weakness, lethargy and impaired appetite which lasted for 72 to 96 h. The sites of inoculations showed varying degree of inflammatory reaction during the first two to three weeks of infection. All experimentally inoculated goats except one developed lesions typical of caseous lymphadenitis in regional/local lymphnodes within 21 days post-inoculation. Route of infection did not influence the ability to set up lesions in lymphnodes. Although massive dose of bacterial (1.2 x 107) was administered, none of the goats had fatal infection indicating that goats are relatively resistant to this infection. Majority of goats did not develop generalized form of caseous lymphadenitis as there was no lesions in visceral/deep seated lymphnodes or organs. The normal serum protein concentration of cross-bred Malabari goats was estimated to range from 7.187 to 9.750 g %. Consequent to experimental infection, serum protein concentration was increased and recorded significant rise from the 5th week onwards reaching the peak value by the 8th week – 11.346 g%. Estimation of quantitative distribution of serum protein fractions was done by agar gel electrophoresis and densitometer tracing of electrophoretogram. Though there was initial increase in globulin content in infected animal followed by a decrease, no significant alteration in the albumin-globulin ration (A:G ratio) was noted compared to the control group. C. psuedotuberculosis was cultivated in lemco proteose broth containing sheep serum and incubated aerobically at 370C for 72 h. Supernatant obtained from the above culture, having maximum haemolysin titre and dermonecrotoxicity was used as the toxin of the bacterium in the present studies. The haemolysin content of the culture supernatant was estimated by the haemolysis test using sheep red cells. A maximum titre of 1:256 was found in the culture aged 72 h. The dermonecrotoxicity of the toxic culture supernatant was tested by intradermal inoculation into the rabbit skin. The inflammatory and necrotic reactions were maximum by 48 h post-injection. Specific antibody activity against toxin of C. pseudotuberculosis in the serum was monitored by haemolysis inhibition test and the test was adjudged as a useful test for detecting humoral immune response to C. pseudotuberculosis infection in goats. In infected goat from the 3rd week of infection onwards MIT was positive while it was negative in control goats during the period of 13 weeks of observation. The peak antibody level was achieved by the 5th week of infection and thereafter the titre was found to dwindle gradually till the 11th week. Towards the end of the observation period (12th week) there was a marginal increase in the antibody titre, which would be considered as secondary immune response against the toxin of the multiplying bacteria. Infected goats showed leukocytosis during the entire period of observation and maximum leukocytosis was observed during the 2nd week of infection. The periodical fluctuation in leukocytosis indicated the recurrent flare up of bacterial invasion in the body. The absolute lymphocyte count obtained both at pre and post-infection periods with experimentally infected goats did not show any change which indicated no deleterious effect on peripheral blood lymphocytes. Throughout the period of observation infected animals showed numerically low lymphocyte percentage in differential counts and with several samples the percentage distribution was significantly low. Absolute counts of neutrophils were consistently high in experimental goats when compared to those of controls and the same was reflected in differential count also. The other blood cells were absolutely without any change in infected as well as control goats. Density gradient centrifugation using Ficoll-paque (1.077 g/ml, centrifuged at 720 x g) was found quite useful for separation of mononuclear leukocytes from the whole blood of goats. Such separated mononuclear cells were found to contain on an average 91.80% lymphocytes and 8.2% monocytes with an average viability of 91.2%. Peripheral blood B-lymphocytes of goats were successfully enumerated by EAC rosette assay employing bovine red cells. The normal percentage of B-cells was estimated to range 8.56 and 12.3. Significantly high percentage of B-cells was recorded in infected animal from the 2nd to 10th week post- infection except at the 5th week. B-cell percentage in infected goats ranged between 12.3 + 0.85-17.63 + 1.2 while it was 8.56 + 0.75 to 12.3 + 1.09 in control goats indicating the operation of humoral immune response to concurrently boost the specific antibody activity in the serum. T-lymphocytes of goats were identified and enumerated by E-rosette assay and ANAE activity. Goat lymphocytes presented several receptors to sheep red cells, as majority of rosettes presented erythrocytes at the entire periphery of lymphocytes. The mean percentage of E-rosette positive lymphocytes in the peripheral blood of control goats ranged from 24.55 + 3.66 to 26.74 + 1.34 during 13 weeks of observation. The E-rosette technique employed in the present study was assumed unaffected by unknown variables as the data recorded in the control goats remained near normal throughout the observation period. During the first two weeks of infection E-rosette positive lymphocyte count was found numerically decreased and the reduction was significant at the first week (18.44 + 1.40). From the third week onwards an increase in the E-rosette forming cells was observed and significant increase was noted during the 5th, 6th, 7th, 8th, 12th and 13th week of infection, the maximum being at the 13th week (35.24 + 1.58). T-cells were also identified and enumerated based on the demonstration on ANAE activity. Fixing of mononuclear cells in acetone-citric acid solution enabled the fixed smears to be stored in dry state without any interference to the enzymic activity for longer periods. T-lymphocytes presented one or two localized red coloured reaction product in the cytoplasm adjacent to the cell membrane. Mean percentage of ANAE positive cells in experimental goats was 28.09 + 1.51 while it was 35.80 + 4.86 for control goats when estimated before the start of the experiment. During infection, the count of ANAE positive cells in the peripheral blood did not show any change as the mean percentage ranged between 28.9 + 2.06-33.78 + 1.99 as against the corresponding values (30.83 + 3.5-36.91 + 3.61) in controls. In infected animals significant hike in E-rosette positive lymphocyte counts was recorded while such a change could not be observed with ANAE positive lymphocytes. Thus the results of T-cell estimation by E-rosette assay and ANAE demonstration indicated that estimation of total rosette forming cells could reflect better the T-cell competence. Cell mediated immune response to C. pseudotuberculosis infection in goats was demonstrated by leukocyte migration inhibition test under agarose. A population density of 1.5 x 108 leukocytes/ml was found suitable for LMIT. Toxic culture supernatant having haemolysin titre 1:16 whose pH adjusted to 7.2 could be successfully used as antigen in the test. In experimentally infected goats leukocyte migration index was less than 0.8 during post-infection period while with control goats it was above 0.8. Significant reduction in LMI index was noted by 45th day of infection through 75 days showing maximum reduction by the 60th day. Intradermal injection of toxic culture supernatant elicited characteristic delayed type skin hypersensitivity reaction in all experimentally infected goats, while a negative reaction in controls. Skin hypersensitivity reaction was found to be maximum by 48 h post-injection. Histopathology of skin biopsy taken from the site of inoculation revealed infiltration of lymphocytes, and macrophages at perifollicular and periglandular areas, congestion of blood vessels with perivascular infiltration of lymphocytes and macrophages and dermal oedema. Tuberculin failed to produce a positive skin hypersensitive reaction in C. pseudotuberculosis infected or control goats. From 15th day onwards, experimentally infected goats which were necropsied presented gross lesions typical of caseous lymphadenitis in lymphnodes. The lesions were found to confine to superficial lymphnodes adjacent to the site of inoculations. The histological changes observed in lymphnodes were basically of two types: hyper-plastic stimulatory reaction and degenerative changes. The changes were hyper-plastic reactive follicles with well distinguished germinal centre, accumulation of lymphocytes and varying degrees of sinus histiocytosis in medullary region, dense lining of medullary cords with plasma cells, depletion of lymphocytes from the cortical area; subcapsular and cortical oedema, congestion of blood vessels, haemorrahage, infiltration of mononuclear cells in lymphatics and blood vessels, accumulation of macrophages and plasma cels in the medulla, dilatation of sinusoids, fibrous tissue proliferation, degenerative and necrotic changes of lymphocytes in the cortex and medulla, fibrous tissue encapsulated focal areas of caseation of calcification surrounded by lymphocytes, macrophages and giant cells and finally conversion of parenchyma to a caseated mass enclosed in fibrous tissue capsule. In brief, the results obtained from the present study revealed the operation of both cell-mediated and humoral immune responses in goats against C. pseudotuberculosis infection. Of the various methods employed to monitor the immune response, leukocyte migration inhibition and delayed skin hypersensitivity tests were suitable for ascertaining the cell-mediated immune response and haemolysis inhibition test for humoral immune response. Leukocyte migration inhibition test and haemolysis inhibition test can be successfully employed for the early diagnosis of C. pseudotuberculosis infection in goats.
Open 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.
Open Access
Prevalence of chlamydial agents in livestock in Kerala
(Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 1988) Reji Francis; KAU; James, P C
The magnitude of the prevalence of chlamydial infections in the live stock in Kerala was assessed by screening the smears of various clinical materials after staining , isolation using chicken embryos and guinea pigs and serologically by passive haemagglutination test. The results obtained were discussed correlating to the managemental practises in the organised herds and agroclimatic conditions. A total of 71 bio sample comprising 17 bovine abortion materials , 15 bull semen, 6 seminal vesicular secretion , one synovial, fluid from a calf, 5 caprine abortion material, 15 caprine pneumonic lungs, 5 samples from perinatal mortality in kids , 4 ovine lung tissue and one conjunctival washings from a buffalo were utilised for screening the smears stained by Gimenez ,Macchiavellos, modified Ziehi-Neelsen and Giemsa's methods and for isolation purpose . On screening the stained smears, one bovine abortion material, two bull semen, one caprine abortion material and three caprine pneumonic lesions were found positive for developmental forms of chlamydiae which could be discerned intra and extracytoplasmically . The overall prevalence rate by this method was 9.9% and species wise prevalence rates were 7.3 % among cattle and 16 % among goats.. Attempts for isolation resulted in the recovery of chlamydiae from two of 17 bovine abortion materials , four of 15 bull semen and two of 15 caprine pneumonic lungs. The overall prevalence based on isolation rate was 11.3% and species wise prevalence rates were 14.6% and 8% respectively for cattle and goats. A total of 169 serum samples consisting of 92 from cattle , 67 from goats and 10 from sheep were screened serologically . Of these 21 from cattle , 13 from goats and one from sheep were found positive showing titres of 1:16 and above . The overall seroprevalence rate was found to be 20.7 % and species wise rates were 22.8%, 19.4% and 10% for cattle , goat and sheep .
Open 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.
Open 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
Open 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.
Open 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.
Open Access
Investigation on the aetiology of plague -like disease in ducks In Kerala
(Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 1978) Krishnan Nair, G; KAU; Sulochana, S
An investigation was carried out to isolate, characterize and identify the agent responsible for the outbreak of duck plague – like disease in ducks in Kerala. Specimens (liver and spleen) from field cases, were processed for virus isolation and were inoculated into either developing duck or chick embryos, by chorio – allantoic (C.A.M.) or allantoic cavity method. Virus isolation was possible only by C. A. M. inoculation of duck embryos and was confirmed by inoculation of the C.A.M. extracts into duck embryo fibroblast (D.E.F.) cell cultures. The cytopathic changes produced by the field isolate DPV – N; its physico – chemical characteristics such as sensitivity to chloroform and 5 – iodo – 2 deoxyuridine; and the effect of exposure to various pH values such as 4.7, 7.2 and 9.1, were compared with that of a known duck plague virus DPV – K, received from the Veterinary Biological Institute, Mannuthy. In D.E.F. cell cultures, the cytopathic changes produced by DPV – N and DPV – K were rounding and clumping of cells, with characteristic basophilia and granulation of the cytoplasm. Although the initial titers of both DPV - N and DPV - K were only 105 and 106.25, they increased to 107.5 and 108.25 respectively, on further passages. The field isolate DPV – N and the known duck plague virus DPV – K were sensitive to 5% chloroform, with complete inactivation in ten minutes. Similarly, both the strains failed to multiply and produce cytopathis changes in cells treated with IUdR, at the rate of 100 micrograms per ml. However, differences were observed in their thermostability and pH sensitivity. Although DPV – K was inactivated completely at 560 C. in 30 minutes, DPV – N was only partially reduced in titer. DPV – N was also found to be resistant, when both the strains were exposed to pH 4.7, for a period of four hours at room temperature. But both were unaffected at pH 7.2 and got inactivated at pH 9.1. Both the strains also failed to produce any haemagglutination reaction with chicken R.B.C or precipitation reaction in agar gels. Although duck plague specific antiserum neutralized homologous strain DPV – K and the newly isolated strain DPV – N, the serum titers obtained with the latter was only less. Experimental infection studies have shown that one to six week – old ducklings were equally susceptible to DPV – N and DPV- K, either with the spleen extract or with tissue culture passaged sample. The symptoms and lesions produced in both cases, were similar to those described for duck plague and also to those seen during the disease outbreak in Kerala. The virus that caused an outbreak of duck plague - like disease in Kerala is found to be indistinguishable from that of duck plague. It is also strongly felt that the lack of complete protection of birds vaccinated with duck plague vaccine is due to t possible strain variation between the classical duck plague virus DPV – K and the virus as it occurred during this outbreak. However, it needs thorough in vitro cross neutralization and in vivo cross protection tests before any definite conclusions can be made on the strain variation of duck plague virus.

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