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Affinity Pattern of Pasteurella Multocida Infection in the Different Organs and Tissues in White Leghorn Chickens

S. Hossain
Veterinary Surgeon, Chatkhil, Noakhali, Bangladesh

M. T. Rahman
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh - 2202, Bangladesh

S. Saha
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh - 2202, Bangladesh

K. A. Chouhury
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh - 2202, Bangladesh

P. M. Das
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh- 2202, Bangladesh.

Abstract/ Executive Summary:

Dynamics of virulent avian isolates of Pasteurella multocida infection was studied in experimental avian model following inoculation via intramuscular (IM) and oral (PO) route so that their distribution pattern in blood, liver, kidney and brain can be studied at definite time interval. Distribution pattern was found more rapid in IM inoculated group. Bacteriemia appeared in 3 hours post-inoculation in IM group while 6 hours post-inoculation in PO group. The concentrations of P. multocida in different organs were low in case of oral route of inoculation in comparison to that of IM. The recovery of organisms from different organs in case of PO route of inoculation was low, indicating a low frequency of colonisation in the digestive tract of chickens. On organ basis liver was found more suitable for the growth and multiplication of P. multocida while brain tissues were less affected. The progressive nature of bacteremia suggested that P. multocida was able to resist the bactericidal activity of serum and replicate rapidly in tissue and in the blood stream. Highest concentration of bacteria in blood indicated that the intracellular multiplication was higher than intravascular multiplication.

Keywords: Tissues, White Leghorn Chickens, Pasteurella multocida

Location: Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh

Start Date:

End Date:

Program Area: Animal Health and Management

Commodity: Chicken

Objectives:

To determine the concentration of P. multocida in blood, liver, kidney and brain at definite intervals following intramuscular and oral administration and their role in the development of the disease.

Methodology:

A stock culture of P. multocida ( PM - 38 ), serotype I ( X - 73 ) was obtained from the repository of the Department of Microbiology and Hygiene, Bangladesh Agricultural University ( BAU ), Mymensingh to use in the experiment. The bacteria were originally isolated from a dead chicken of a outbreak area of BAU Poultry Farm. The strain was made virulent by serial passages through blood agar, mice and chickens. A total of 93, three months old White Leghorn ( WLH ) chickens of either sex were collected from BAU Poultry Farm to use in the experiment. The birds had no previous history of infection or of vaccination against Fowl cholera. They were kept in poultry cages for 7 days to acclimatise with the new environment and to observe abnormal behaviour, if any. Sufficient balanced ration and water were supplied ad libitum to the chickens. The birds were randomly divided into major three groups : Group A consisting of 24 birds, Group B consisting of 24 birds and Group C consisting of 36 birds. The remaining 9 birds were used to enhance the virulence of the organisms. Specific pathogen free healthy Swiss albino mice ( 6 weeks old ) were obtained from the experimental animal house of the Department of Microbiology and Hygiene, BAU, to increase the virulence of the bacteria. The animals were kept in an isolated room for the observation of clinical signs and symptoms after inoculation. Sufficient feed and water were supplied to the animals. Blood agar media, mice and chickens were used for enhancement of virulence from the stock culture P. multocida was allowed to grow in blood agar ( BA ) media and incubated at 37°C. After 24 hours, a single medium size smooth colony from plate was taken and suspended in 5 ml of PBS. Thereafter, three serial passages ( three birds in each passage) were made in mice via subcutaneous ( SC ) route. From the third passage, further three serial passages were made to the chickens. All the birds of third passage died within 18 to 24 hours of inoculation and considered that the virulence was enhanced. In all the steps, sterility tests were conducted. Liver samples were collected from the infected birds that died at 18 to 24 hours post- inoculation ( PI ) of their 3^rd passage and cultured of BA plates. One loopful bacterial colony from 18 hours old pure culture was diluted 10-folds in PBS. A 0.5 ml of bacterial suspension from 105 dilution was used as inoculum, at the same time the colony forming unit ( CFU ) of the respective dilution were determined by plate count method and inoculum of approximately 4.5 x 10⁵ CFU / 0.5 ml was used in the experiment. Each bird of Group A received 0.5 ml inoculum ( approximately 4.5 x 10⁵ CFU ) of P. multocida intramuscularly and each bird of Group B received similar number of bacteria via oral route. Each bird of control group was inoculated either by intramuscular ( IM ) or oral route with sterile phosphate buffer saline as mock antigen. Inoculated birds were observed closely at 3, 6, 12,48 and 96 hours post-inoculation for clinical signs, if any. At 3, 6, 12, 24, 48 and 96 hours PI blood were collected from each bird to determine the bacterial load. Three birds from each group were randomly selected at the aforesaid time intervals and euthanesised to find out the bacterial load in different organs. Immediately after collection of I ml of blood from each bird, it was placed in 9 ml of PBS. Samples from each organ ( liver, kidney and brain) was weighed, grinded by pestle and mortar and suspended in PBS and then homogenised individually. A serial la-folds dilution was made with tissue homogenates or blood samples with PBS and the CFU was determined by standard plate count method. The liver, kidney and brain were collected aseptically and individually on sterile petridishes for bacteriological examination. Impression smears from the heart blood, liver, kidney and brain of the birds were stained with Leishman's and Gram's stain following the procedure of Macleod to observe the bipolar and capsular P. muliocida.

Source of Information: Bangladesh Vet. J. (1998).32 (3-4): 118 - 123

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In the present study, it was observed that the concentrations of P. multocida in different organs were low in case of oral route of inoculation in comparison to that of IM. The recovery of organisms from different organs in case of oral route of inoculation was low, indicating a low frequency of colonization in the digestive tract of chickens. There are some barriers present could be the IgA, which is normally available in the tract. IgA probably provides an important barrier in the mucosal epithelia, giving a first line of defense against pathogens. In bacterial septicaemic diseases, such as Fowl cholera, the usual stages of disease development are adhesion, colonization, invasion, multiplication and host damage. Colonization occurred in less than 6 hours after exposure and invasion occurred between 6 and 12 hours in case of upper respiratory tract exposure.

Knowledge Management: Journal