Theses

Browse

Search Results

Now showing 1 - 9 of 18

Open 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.
Open 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.
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
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
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.
Open Access
Polymerase chain reaction for the detection of canine parvovirus in faeces of dogs
(Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 2004) Josemi, Mathew; KAU; Mini, M
A study was undertaken to compare the ability of haemagglutination (HA), polymerase chain reaction (PCR) and seminested PCR for the diagnosis of canine parvovirus (CPV) infection by detecting CPV from faecal samples of clinically suspected dogs. Characterization of vaccine strain and field strains of CPV was performed by restriction enzyme analysis (REA) in this study. Attempts were made to isolate CPV from faecal samples in MDCK cell line. One hundred and twentysix faecal samples were collected from dogs suspected for CPV infection and 40 faecal samples were also collected from normal healthy non-vaccinated as well as vaccinated dogs after 15 days of vaccination, that were brought to veterinary hospitals attached to KAU. All the samples were screened by HA, PCR and seminested PCR to detect CPV. No significant difference in HA titre could be appreciated on comparing the titre using PBS, pH 7.2 and PBS – BSA as diluents. The haemagglutination reaction by CPV was found to be favoured by a slightly acidic pH in the range of 4.0 to 6.0. Chloroform treatment of faecal samples had no influence on HA titres above 5 log2. Among 126 faecal samples screened, 43.65 per cent, 61.11 per cent and 73.81 per cent were tested positive for CPV infection by HA, PCR and seminested PCR respectively. Therefore, seminested PCR was found to be a more sensitive and specific method over HA and PCR for the early diagnosis of CPV infection. All the faecal samples from healthy non-vaccinated and vaccinated dogs after 15 days of vaccination were tested negative by HA, PCR and seminested PCR. Restriction enzyme analysis using HinfI, Rsa I and Sau 961 revealed no difference in the fragment length patterns between CPV vaccine strain and field strain. Attempts to isolate CPV from faecal samples in MDCK cell line were found unsuccessful. The occurrence of CPV infection among non-vaccinated dogs was found to be higher than that of vaccinated dogs. Vaccine failure was also observed after MLV vaccination. The distribution of CPV infection was highest among dogs between two to four months of age. Breed-wise distribution of CPV infection showed highest distribution in German shepherd dogs when compared to other breeds. Most of the cases of CPV were noticed during February and July in the year 2003 and during March to June in the year 2004.
Open Access
Molecular methods based detection of pathogenic mycoplasmas of chicken
(Department of Veterinery Microbiology, College of Veterinary and Animal Sciences, 2006) Dipu, M K; KAU; Jayaprakasan, V
A study was undertaken for the detection of Mycoplasma DNA from the specimens by Polymerase Chain Reaction (PCR), differentiate the three significantly pathogenic mycoplasmas of chicken namely M. gallisepticum, M. synoviae and M. iowae from the other less pathogenic ones based on the result of polymerase chain reaction. Attempts were also carried out to isolate Mycoplasma from clinical samples testing positive by polymerase chain reaction and from a few randomly selected negative samples and to comprehend the different strains of M. gallisepticum, if any, among chicken of various age groups. Out of a total of 225 birds subjected for the study 25 were found positive for the presence of avian Mycoplasma by genus-specific PCR. Thirty samples from these twenty-five birds were positive. Tracheal swabs form all these birds were positive. Fifteen isolates were obtained from these twenty five birds when the tracheal swabs were directly streaked onto BHI agar whereas when these tracheal swabs were collected in BHI broth initially and later subcultured onto BHI agar following a positive result in PCR / colour change of the broth, only 13 of them yielded colonies. Five of the isolates were found to be M. gallisepticum by MG-PCR and these were obtained upon direct inoculation of BHI agar with tracheal swabs from birds. Those samples collected in BHI broth yielded MG colonies only in two cases. Non-MG colonies were found interspersed with MG colonies in agar plates and thus the selective isolation of MG colonies must be performed prior to subculture. The utility of PBS and buffered peptone water supplemented with glycerol as transit media for clinical samples intended for PCR was evidenced. The isolates obtained were successfully lyophilized and stored at –70°C throughout the span of the study. The PCR-RFLP pattern of the obtained MG isolates revealed uniformity among the isolates. None of the samples yielded positive result in MS-PCR and MI-PCR. The sensitivity and usefulness of molecular methods based detection of pathogenic mycoplasmas of chicken over isolation techniques could be appreciated.

Theses

Browse

Thesis Type

Type

Author

Date

Subject

Search Results