ISOLATION AND IDENTIFICATION OF SALMONELLA FROM FAECAL SAMPLES OF DIFFERENT AGES OF SNAKES

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ISOLATION AND IDENTIFICATION OF SALMONELLA FROM FAECAL SAMPLES OF DIFFERENT AGES OF

SNAKES

SUMMARY

A cross-sectional study was conducted to investigate the prevalence of Salmonella in fecal samples of snakes at Sitakund and Foy’s Lake area of Chittagong district. In total 25 samples that were collected between August 2008 to March 2009, were subjected for the screening of Salmonella using the conventional cultural procedures with SS agar. Of those samples, 18 samples were positive which form 72% of total collected samples. The study reveals that salmonellosis is more in large population areas where snake boxes were in close contact. The study also reveals that salmonellosis somewhat affects in mid age. During the collection of samples all the snakes were healthy which is normal phenomenon for most of the reptiles.

Key words: Salmonellosis, Snake, S.S. Agar, Zoonosis.

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CHAPTER-I

INTRODUCTION

In Bangladesh, reptile farming is quietly a new application. Now there is an established crocodile farm in Mymensing. Another reptile, snakes are reared from ancient ages for various purposes. Snake charmers rear snakes mainly for livelihood by showing play. Some charmers took attempt to establish an organized snake farm but till they could not make it possible. In near future there has a chance to establish a snake farm in our country. Now the snakes that are reared in captivity are mostly belongs to snake charmers’ house and rest are in the zoo. But, many people such as veterinarians, more charmers, Herpetologists, researchers will involve in near future. As a result, they will be at risk from reptilian zoonosis.

Among the reptilian zoonoses the most recognized one is salmonellosis.

Salmonella spp. are gram negative straight rods, usually flagellated, facultative anaerobes (Krieg et al,1984). There are over 2000 serotypes within the species S.enteritidis (e.g.,S.enteritidis serotypes typhimurium), while S. typhi, causative agent of typhoid fever and S. choleraesuis have only one serotype each. The classification system is usually abbreviated and the enteritidis is deleted and the name S. typhimurium is applied. Salmonellae are pathogenic to a variety of animals and humans(Fox et al,1991).

The zoonotic significance of reptilian salmonellosis was first reported in 1963 when a 7-month old infant acquired the disease from a pet turtle, although the potential had been recognized since 1946 when

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Salmonella spp were originally isolated from turtles(Lamm et al,1972).

So, there is a greater chance to be infected with salmonellosis from other reptile source such as snakes. This study aims to give the overall prevalence of Salmonella in feces of various species of 25 snakes of various ages to prove the presence of salmonellae in subclinical form that may lead to some sort of zoonotic diseases.

Objectives

1) To determine the prevalence of Salmonella in feces of snakes by collecting them from the snake boxes from the snake charmers’ house.

.

2) To identify the Salmonella isolates in feces that do not show any clinical sign to the snake host.

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CHAPTER-II

LITERATURE REVIEW

2.1 Snake

Background

in Indian Sub-continent:

Snakes are ectothermic air breathing vertebrates, covered with scales or plates with no hair, feather or mammary gland. Snakes are widely distributed in the Indian sub- continent from seas to near the snow-lines in Himalayas, comprising of 216 species of snakes, including 52 species whose bite may cause some toxic symptoms in man and animals. Considering the importance of snakes in venom production and their ecological role in controlling the rodent population and myths in the Indian culture, conservation of snakes is given much emphasis in the recent period(Rajesh,2007).

2.2 What is

Salmonella

?

Salmonella is a genus of rod-shaped Gram negative enterobacteria that causes typhoid fever, paratyphoid and foodborne illness. Salmonella does not ferment lactose. It is motile in nature and produces hydrogen sulfide.

Salmonellae are ubiquitous in human and animal pathogens, and salmonellosis, a disease that affects an estimated 2 million Americans each year, is common throughout the world. Salmonellosis in humans usually takes the form of a self-limiting food poisoning (gastroenteritis), but occasionally manifests as a serious systemic infection (enteric fever) which requires prompt antibiotic treatment. In addition, salmonellosis causes substantial losses of livestock.

A strain of bacteria that can enter the human system through contaminated water or food such as meat or poultry, and eggs with cracked shells. Other foods can be contaminated by touching salmonella- carrying foods or unwashed surfaces (like cutting boards) that have had contact with them. The presence of Salmonella is difficult to detect because it gives no obvious warnings (such as an off smell or taste). The

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bacteria can cause stomach pain, nausea, vomiting, diarrhea, headache, fever and chills. It can attack in as little time as 6 to 7 hours or take as long as 3 days. It seldom causes death and can be cured with antibiotics.

Any of various rod-shaped bacteria of the genus Salmonella, many of which are pathogenic, causing food poisoning, typhoid, and paratyphoid fever in humans and other infectious diseases in domestic animals. (The American Heritage Dictionaries)

Any of the rod-shaped, gram-negative, non-oxygen-requiring bacteria that make up the genus Salmonella. Their main habitat is the intestinal tract of humans and other animals. Some of the 2,200 species exist in animals without causing disease; others are serious pathogens. Any of a wide range of mild to serious infections caused by salmonellae is called salmonellosis, including typhoid and paratyphoid fever in humans.

Refrigeration prevents their reproduction but does not kill them; as a result, many salmonellae can develop in foods, which, when eaten, can cause gastroenteritis. Chickens are major reservoirs of Salmonella, and chicken and eggs are the principal source of human poisoning, whose symptoms include diarrhea, vomiting, chills, and painful headaches.

Other food sources include unpasteurized milk, ground meat, and fish.

(British Concise Encyclopedia)

A category of bacteria that occurs in many pathogenic forms. One kind causes typhoid fever; there is evidence that other kinds cause various forms of food poisoning. (Haughton Mifflin Company)

A genus of gram-negative, non-lactose fermenting, medium-sized, rod- shaped, bacteria, members of the family Enterobacteriaceae, most species having flagella and pili. The genus contains one species which has been divided into seven subgroups and a very large number of serotypes. Most species pathogenic for warm-blooded animals are in subgroup I (S. enterica). Subgroups IIIa (S. salamae) and IIIb (S.

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arizonae) include some species occasionally pathogenic for animals and birds. The Salmonella include the typhoid and paratyphoid bacilli and bacteria usually pathogenic for lower animals but which are often transmitted to humans. They cause salmonellosis which has a number of manifestations and some are specific causes of abortion. (Elsevier Veterinary Dictionary)

A genus of aerobic gram-negative rod shaped nonspore-forming usu.

Motile bacteria of the family Enterobacteriaceae that grow well on artificial media and form acid and gas on many carbohydrates but not on lactose, sucrose, or salicin, that are all pathogenic for man and other worm-blooded animals, and that are chiefly associated with various types of food poisoning, with acute gastrointestinal inflammation (as in typhoid fever), or with diseases of genital tract. (Webster’s Medical Dictionary).

2.3 History

Salmonella was named after Daniel Elmer Salmon, an American veterinary pathologist, although it was his partner and contemporary Theobald Smith (better known for his work on anaphylaxis) who first discovered the bacterium in 1885 from pigs.

The zoonotic significance of reptilian salmonellosis was first reported in 1963 when a 7-month old infant acquired the disease from a pet turtle, although the potential had been recognized since 1946 when Salmonella spp were originally isolated from turtles (Lamm et al,1972).

2.4 Scientific classification Kingdom : Bacteria

Phylum : Proteobacteria

Class : Gamma Proteobacteria Order : Enterobacteriales

Family : Enterobacteriaceae Genus : Salmonella

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TABLE 1: SEROTYPES ASSOCIATED WITH HUMAN SALMONELLOSIS (`Chiodini et al,1981)

Salmonella blockley S. enteritidis

S. heidelberg

S. infantis

S. javiana

S. montevideo

S. newport

S. saint-paul

S. tennessee

S. thompson

S. typhi

S. typhimurium

TABLE 2: SALMONELLA SEROTYPES IMPLICATED IN CASES OF REPTILE ZOONOSIS (Fujita et al, 1981)

Salmonella agona

S. arizonae

S. berta

S. blockly

S. braenrup

S. brandenburg

S. chester

S. enteritidis

S. give

S. hartford

S. heidelberg

S. infantis

S. java

S. litchfield

S. marina

S. muenchen

S. newport

S. oranienburg

S. overschie

S. panama

S. paratyphi-B

S. poana

S. pomona

S. poona

S. saint-paul

S. san-diego

S. schwarzengrund

S. Thompson

S. typhimurium

S. urbana

S. wassenaar

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2.5

Characterization of the organism

Salmonella species are Gram-negative, flagellated, nonsporogenic, nonmotile, facultative anaerobic. They are slender rods, 1.0-2.5 µm in length and 0.3-1.5 µm in width. The bacilli mostly occur singly, but occasionally two or more can be found united. (Shivaprashad, 1997).

There are over 2300 serologically distinguishable variants (Gast, R.K., 1997).

Antigenic characters

1) Somatic or O antigens: designated by Arabic numerals; group classification is based on several of these antigens.

2) Flagellar antigens-phase 1: Designated by small letters of the alphabet, more or less specific for Salmonella. Isolates must be in this phase before they can be typed. Phase 2: Designated by Arabic numerals;

less specific, and duplicated in other bacterial species.

3) K antigen (capsular or envelop): “Vi” antigen, “M” antigen and so forth. These antigens may interfere with agglutinability of O antisera.

(Carter G.R. et al., 1995)

Serotyping

The primary species of Salmonella is called Salmonella enterica. Over the past decade, Salmonella bongori has become the second bona-fide species, which has been upgraded from subspecies status seen in the ninth edition of Bergey's Manual of Determinative Bacteriology (1994).

Subspecies are defined and differentiated biochemically and genetically.

Also termed "subgroups," the subspecies names in the table are preceded by the numerical subgroup designation.

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Several host-adapted, biochemically aberrant serovars ("bioserovars") which do not have unique antigenic formulae (and must then be distinguished by their biochemical reactions) are shown in this table – i.e., Pullorum, Gallinarum, Paratyphi-C, Cholerae-suis and Typhi-suis.

Irrespective of subspecies designation, the serovars are grouped according to their O antigens: Those which share O antigen 2 are in Group A, those which share O antigen 4 are in Group B, those which share O antigens 6 and 7 are in Group C1, etc.

Lists of serovars can be found in Bergey's Manual of Systematic Bacteriology, Second Edition, Volume 2, Part B (2005) and the 1998 edition of the Difco Manual. For the ultimate list according to a controlling authority, the WHO (World Health Organization) Collaborating Centre for Reference and Research on Salmonella periodically publishes the Antigenic Formulas of the Salmonella Serovars.

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Subspecies Antigenic formula Serovar name

1. subsp. Enterica 1,2,12:a:1,5 Paratyphi-A

1,4,5,12:b:1,2 Paratyphi-B 1,4,5,12:i:1,2 Typhimurium

1,9,12:-:- Pullorum

1,9,12:-:- Gallinarum

1,9,12:g,m:1,7 Enteritidis

1,9,12:g,p:- Dublin

6,7:c:1,5 Paratyphi-C

6,7:c:1,5 Cholerae-suis

6,7:c:1,5 Typhi-suis

6,7:y:e,n,z15:z47:z50 Mikawasima ¹

9,12:d:z66 Typhi

13,23:d:1,7 Grumpensis ²

30:i:e,n,z15 Mjordan ³ 47a,47b:z45:z4,z23:z6 Bere ⁴

2. subsp. Salamae 1,9,12:l,w:e,n,x Daressalaam ⁵

11:b:1,7 (none given)

3a. subsp. arizonae ⁶ 51:z4,z23:- (none given) 3b. subsp. diarizonae ⁶ 40:k:e,n,x,z15 (none given) 4. subsp. Houtenae 11:z4,z32:- (none given)

43:z4, z23:- Houten ⁵

5. subsp. bongori ⁷ 48:z35:- Bongor ⁵

6. subsp. Indica 1,6,14,25:a:e,n,x Ferlac ⁵

1 An example of a quadriphasic serovar.

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2 An example of a serovar named after the individual from which it was isolated first – in this case a person (hospital patient?) who was referred to as being grumpy (or perhaps as Grumpy). "Grumpensis" is Latin for

"from Grumpy."

3 An example of a serovar named to honor a specific individual – in this case, basketball legend Michael Jordan. The name was seen in the press as Salmonella mjordan.

4 An example of a triphasic serovar.

5 Names for serovars other than those in the first subspecies have been deprecated, and only the antigenic formulas are recognized as serovar designations; the old names for these particular serovars are shown on this table.

6 Set apart in some older classifications as a separate genus (Arizona) or as a separate species of Salmonella (S. arizonae). Subgroup 3a serovars are monophasic; subgroup 3b serovars are diphasic.

7 The sole subspecies of Salmonella bongori.( Laboratory Manual for the Food Microbiology Laboratory (1998 edition, edited by John L. and published at the University)

The classification of salmonellae has been controversial for many years.

According to the latest nomenclature, which reflects recent advances in taxonomy (Popoff M.Y. 2001)., the genus Salmonella consists of only two species: S. enterica and S. bongori (Le Minor L. & Popoff M.Y.

1987, Popoff M.Y., Bockemuhl J. & McWhorter-Murlin A. 1994).

Salmonella enterica is divided into six subspecies, which are distinguishable by certain biochemical characteristics and some of which correspond to the previous subgenera. These subspecies are:

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Previous nomenclature Current nomenclature

Subspecies I Subspecies enterica

Subspecies II Subspecies salamae

Subspecies IIIa Subspecies arizonae

Subspecies IIIb Subspecies diarizonae

Subspecies IV Subspecies houtenae

Subspecies VI Subspecies indica

2.6 Isolation of Salmonella sp.

Numerous techniques and methods have been described for isolation of Salmonella from different types of specimens. An isolation technique for Salmonella in one type of sample may not be suitable for the isolation from other sample types (Fricker, 1987). Therefore, it is not possible to recommend a single technique for isolation of Salmonella. For the special interest of epidemiological investigations with a view to eradicating, controlling or preventing the disease outbreak, detailed strain identification is essential (Threlfall et al., 1994). Identification under the family of Enterobactericae. The isolates under the family of Enterobactericae grow well on artificial media (O.I.E. Manual, 1996).

These Gram negative rods are oxydase negative and catalase positive (Douglas and Alice, 1993). Triple sugar iron agar provides a presumptive test for characterization of 7 salmonellae (Doughlas et al., 1998).

Additional differentiation of Salmonella from other organisms is accomplished by biochemical analysis (Cox and Williams, 1976).

Cultural identification using the SS agar:

Enteric organisms are differentiated by their ability to ferment lactose.

Salmonella spp. and Shigella spp. are non-lactose fermenters and form colorless colonies on Salmonella Shigella Agar. H2S positive Salmonella

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spp. produce black-center colonies. Some Shigella spp. are inhibited on Salmonella Shigella Agar. E. coli produces pink to red colonies and may have some bile precipitation

2.7 Prevalence of Salmonella in Snakes:

Estimates of reptiles harboring Salmonella spp. have been reported to be as high as 83.6 to 93.7 %, depending on the method of testing. Turtles may be infected at a rate of 12.1 to 85

%,snakes 16 to 92% and lizards 36 to 77 % (

Chiodini et al,1981)

. One survey in Canada between 1979 and 1983 tested 150 pet reptiles submitted for necropsy : 51% of the snakes, 48% of the lizards and 7% of the turtles cultured positive for Salmonella spp., with 31 different serotypes idfntified (Onderka et al, 1985).

S. arizonae is more prevalent in snakes, estimated at 78.8%

harboring the organism.

2.8 CLINICAL SIGNS IN SNAKES:

Although the rate of infection is quite high, reptiles usually do

not show signs of disease from Salmonella infection and the

relationship may indeed be a saprophytic

one(Anonymous,1981). Experimental infections in snakes by

oral inoculation resulted in fecal shedding without observable

disease in the host or formation of agglutinins, indicating some

sort of immune response. Infection via subcutaneous,

intracardial or intraperitoneal resulted in the formation of

specific agglutinins without lesion development. In maximum

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cases, highly virulent strains occur in clinically healthy reptiles(

Chiodini et al,1981).

2.9 Culturing Information

SALMONELLA SHIGELLA AGAR (7152) Intended Use

Salmonella Shigella Agar is used for the isolation of Salmonella spp.

and some strains of Shigella spp.

Product Summary and Explanation

Salmonellosis continues to be an important public health problem worldwide. Infection with non-typhi Salmonella often causes a mild, self- limiting illness. Typhoid fever, caused by Salmonella typhi, is characterized by fever, headache, diarrhea, abdominal pain, and can result in fatal respiratory, hepatic, and or neurological damage.¹This infection can result from the consumption of raw, undercooked, or improperly processed foods contaminated with Salmonella spp.

Shigellosis, caused by Shigella spp., is an intestinal illness characterized by abdominal pain, fever, and watery diarrhea. When associated with outbreaks, shigellosis is usually transmitted through contaminated food and/or water.

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Salmonella Shigella Agar is a modification of the Desoxycholate Citrate Agar described by Leifson.²Salmonella Shigella Agar is superior to a number of other media for the isolation of Salmonella spp. and Shigella spp.³Salmonella Shigella Agar is recommended for testing clinical specimens and food testing for the presence of Salmonella spp. and some Shigella spp.^1,4,5

Principles of the Procedure

Beef Extract, Enzymatic Digest of Casein, and Enzymatic Digest of Animal Tissue provide sources of nitrogen, carbon, and vitamins required for organism growth. Lactose is the carbohydrate present in Salmonella Shigella Agar. Bile Salts, Sodium Citrate and Brilliant Green inhibit Gram-positive bacteria, most coliform bacteria, and inhibit swarming Proteus spp., while allowing Salmonella spp. to grow. Sodium Thiosulfate and Ferric Citrate permit detection of hydrogen sulfide by the production of colonies with black centers. Neutral Red is the pH indicator.

Formula / Liter

Beef Extract...5 g Enzymatic Digest of Casein...2.5 g Enzymatic Digest of Animal Tissue...2.5 g Lactose...10 g Bile Salts...8.5 g Sodium Citrate...8.5 g Sodium Thiosulfate...8.5 g Ferric Citrate...1 g Brilliant Green...0.00033 g Neutral Red...0.025 g Agar...13.5 g Final pH: 7.0 ± 0.2 at 25°C

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Formula may be adjusted and/or supplemented as required to meet performance specifications.

Precautions

1. For Laboratory Use.

2. IRRITANT. Irritating to eyes, respiratory system, and skin.

Directions

1. Suspend 60 g of the medium in one liter of purified water.

2. Heat with frequent agitation and boil for one minute to completely dissolve the medium.

3. DO NOT AUTOCLAVE.

Quality Control Specifications

Dehydrated Appearance: Powder is homogeneous, free-flowing, and light to medium pinkish-beige.

Prepared Appearance: Prepared medium is light to medium reddish- orange to peach, trace to slightly hazy.

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Expected Cultural Response: Cultural response at 35°C after 18 - 24 hours incubation.

Microorganism Response Reactions

Enterococcus faecalis ATCC® 29212

complete inhibition

---

Escherichia coli ATCC®

25922

partial to

complete inhibition

pink to rose-red

colonies with

precipitate Salmonella typhimurium

ATCC® 14028

growth colorless colonies with black centers

Shigella flexneri ATCC®

12022

growth colorless colonies

Test Procedure

For isolation of Salmonella spp. and Shigella spp. from clinical specimens, inoculate fecal samples and rectal swabs onto one quadrant of Salmonella Shigella Agar, streak for isolation. Incubate plates at 35°C, and examine after 24 and 48 hours for colonies resembling Salmonella spp. or Shigella spp. Consult appropriate references for food testing.

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Results

Enteric organisms are differentiated by their ability to ferment lactose.

Salmonella spp. and Shigella spp. are non-lactose fermenters and form colorless colonies on Salmonella Shigella Agar. H2S positive Salmonella spp. produce black-center colonies. Some Shigella spp. are inhibited on Salmonella Shigella Agar. E. coli produces pink to red colonies and may have some bile precipitation.

Limitations of the Procedure

1. Salmonella Shigella Agar is highly selective and not recommended as the primary isolation of Shigella. ^1,2,6Some Shigella spp. may be inhibited.

2. A few nonpathogenic organisms may grow on Salmonella Shigella Agar. These organisms can be differentiated by their ability to ferment lactose and other confirmatory tests.

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References

1. P. R. Murray, E. J. Baron, M. A. Pfaller, F. C. Tenover, and R. H.

Yolken (eds.). 1995. Manual of clinical microbiology, 6^thed. American Society for Microbiology, Washington, D. C.

2. Leifson, E. 1935. New culture media based on sodium desoxycholate for the isolation of intestinal pathogens and for the enumeration of colon bacilli in milk and water. J. Pathol. Bacteriol 40:581.

3. Rose, H. M., and M. H. Kolodny. 1942. The use of SS (Shigella- Salmonella) Agar for the isolation of Flexner Dysentery bacilli from the feces. J. Lab. Clin. Med. 27:1081-1083.

4. Isenberg, H. D. (ed.). 1992. Interpretation of aerobic bacterial growth on primary culture media, Clinical microbiology procedures handbook, vol.

1 p. 1.61-1.67. American Society for Microbiology, Washington, D.C.

5. Vanderzant, C., and D. F. Splittstoesser (eds.). 1992. Compendium of methods for the microbiological examination of foods, 3^rded. American Public Health Association, Washington, D.C.

6. Taylor, W. I., and B. Harris. 1965. Isolation of shigellae. II.

Comparision of plating media and enrichment broths. Am. J. Clin. Pathol.

44:476.

7. McFaddin, J. F. 1985. Media for isolation-cultivation-identification- maintenance of medical bacteria, Vol. 1. Williams & Wilkins, Baltimore, MD.

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CHAPTER-III

MATERIALS AND METHODS

Study area and duration

The study continued from August 2008 to March 2009, during my extramural studies (internship) phase. In this phase samples were collected from snakes of snake charmers’ house from Foy’s lake and Sitakunda area of Chittagong district.

Collection of fecal samples

Total 25 samples were collected from 3 snake charmer’s house of which 12 samples from Md. Salim of Foy’s Lake, 8 from Sumon Shil of Foy’s Lake and 5 from Abul Kashem of Sitakund of Chittagong district. For collection of sample each snake was fed fishes and then fecal samples were collected on 3

^rd

to 5

^th

days depending on time of defecation of snake after taking feed.

Then the samples were kept into the refrigerator at 3-4

⁰ C

at the

Microbiology Laboratory of Chittagong Veterinary and Animal

Sciences University and the samples were stored there until

further study was conducted.

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Fig 1. Some collected samples before storing showing polythene bags wrapped over.

Isolation and identification of Salmonella from the faecal samples.

The isolation and identification of Salmonella in the feacal samples was carried out on SS agar. Briefly, 1g of each of the collected faecal sample was diluted into 10 ml of PBS and then the diluted samples were centrifuged at 1000 RPM for 10 minutes. After that from supernatant of each centrifuged sample individual streaking was done on separate Petri dish filled with SS agar. After streaking, the Petri dishes were kept at 37 ⁰ C for 48 hours. Then the colonies are observed for identification of Salmonella.

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Fig 2. SS agar plate, PBS and Samples diluted with PBS

Fig 3. Streaking on SS agar

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Fig 4. Petri Dishes with SS agar after streaking on incubator

Fig 5. Observing growth characteristics of an inoculated samples on a SS agar plate

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CHAPTER-IV

RESULTS

Table-3 presents the growth characteristics (either positive or under study negative) of bacteria grown from 25 fecal samples of snakes.

Overall, of the 25 samples investigated, 18 were found positive with Salmonella; 9 of them were isolated from 12 samples of Md. Salim, 6 from 8 samples of Sumon Shil and 3 from 5 samples of Abul Kashem.

Statistically, 72% of the collected samples was positive. The fecal sample shows that 75% of the samples was positive from Md. Salim, 75% from Sumon Shil and 60% from Abul Kas

hem.

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Table 3. Growth properties (either positive or negative) of the bacteria grown from 25 fecal samples of snakes under study

Name of Snake

Charmers

Species of Snakes Age of Snakes

Positive or negative on SS agar

Md. Salim 1.Asiatic Rock Python 2m +

3y +

2.Spectackled Cobra 5y -

4y +

10y +

3.Monocled Cobra 7y -

11y +

4.Banded Krait 7y +

5.Rat Snake 9y +

6.Golden Flying Snake 4y -

7.Common Vine Snake 5y +

3y +

Sumon Shil 1.Asiatic Rock Python 7y +

2.Spectackled Cobra 13y +

8y +

3.Monocled Cobra 9y -

6y +

2y +

4.Banded Krait 9y -

5.Golden Flying Snake 4y +

Abul Kashem

1.Copperhead 8y +

9y +

2. Common Vine Snake 3y -

3. Spectackled Cobra 10y -

4. Rat Snake 5y +

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Fig-6.

Prevalence of salmonellosis on different age group

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CHAPTER V

DISCUSSION

Salmonella have both pathogenic and zoonotic importance. From my study it is evident that 72% of the snakes of study are harboring Salmonella, from where humans can be infected.

Because of time and laboratory resource constrain, this study failed to speciate isolates of Salmonella recovered from fecal samples of marketed chickens. Also, the serotyping of them was not done due to lack of biologic resources.

Here it is clear that in case of large population the prevalence is high may be due to less space and more close contact of the snake boxes.

Again, on different age group, there is found that salmonellosis occur mostly between 5 and 8 years of age.

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CHAPTER-VI

CONCLUSION

The results obtained through this short-period of study reveals that about 72% of the collected samples were positive for salmonellosis. This could be hazardous for the public health. Because in most of the positive cases with salmonellosis are in the subclinical form. But when the organism is transmitted to human and other animals, it can produce harmful diseases.

So, peoples who will work later, should be cautious in handling snakes.

Otherwise, diseases may occur from relatively healthy snakes.

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ACKNOWLEDGEMENTS

At the inception, I wish to acknowledge the immeasurable grace and profound kindness of the “Almighty Allah,” the supreme Ruler of the Universe, without Whose desire I could not conclude this report.

The author would like to express his deep sense of gratitude and thanks to Professor Dr. N. C. Debnath, honorable V. C., Chittagong Veterinary and Animal Sciences University, Chittagong.

The author would like to express his deep sense of gratitude and heartfelt appreciation to Professor DR.M.A.Motin Prodhan, Dean, Faculty of Veterinary Medicine, Chittagong Veterinary and Animal Sciences University, Chittagong.

The author extends his gratefulness to Professor DR. Md. Rayhan Faruque, Head, Department of Medicine & Surgery, Chittagong Veterinary and Animal Sciences University, Chittagong for his scholastic guidance and supervision of the report work and writes up of the dissertation.

The author highly express his sincere gratitude and gracefulness to DR. H.

Barua, Assistant Professor, Chittagong Veterinary and Animal Sciences University, Chittagong, for his valuable suggestions and advice.

The author also extends his gratefulness to Zahirul Hoque, Director of Sitakund Computer Academy for giving me assistance to use his computer lab.

This Author is very much grateful to the snake charmers, Salim bhai, Sumon da and Kashem kaka for their cordial help.

The author also extends his gratefulness toDR. M. Murshed , Curator, Chittagong Zoo, Chittagong, for giving me valuable advice and introducing me to the snake charmers.

Finally the author express his good wishes and warmest sense of gratitude to all his friends and families.

The Author

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Anonymous (2006). Annual report on zoonoses in Denmark 2005. Danish Institute for Food and Veterinary Research. Ministry of Family and Consumer Affairs.

Anisuzzaman, M., Wahid, M.A. and Ali, A. 1990. Adaptibility of Fayoumi chickens under different management conditions. Bangl.

J. Anim. Sci. 19: 63-69.

Bhattacharjee, P.S., Kundu, R.L., Mazumder, J.U., Hossain, E. and Miah, A.H. 1996. A retrospective analysis of chicken diseases diagnosed at the Central Disease Investigation Laboratory, Dhaka, Bangladesh. Bangladesh Vet. Jr. 30: 105-113.

Bergey's Manual of Systematic Bacteriology, Second Edition, Volume 2, Part B (2005)

Chiodini RJ, Sundberg JP: Salmonellosis in reptiles: a review. Am J Epidemiol 113:494,1981

Cox, N.A. and Williams, J.E. (1976). A simplified biochemical system to screen Salmonella isolates from poultry for serotyping. Poultry Science. 55:1968-1971.

Douglas, W.W. and Alice, M.H. 1993. Isolation of Salmonella from chickens reacting in the pullorumtyphoid agglutination test. Avian Diseases. 37: 805-810.

Fricker, C.R. 1987. Isolation of Salmonella and Campylobacter. Journal of Applied Bacteriology. 63: 99-116.

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