Shigella spp.

Important Species:

 SerogroupA: S. dysenteriae
 SerogroupB: S. flexneri (9 serotypes)
 SerogroupC: S. boydii (19 serotypes)
 SerogroupD: S. sonnei (one serotype)

Sources:

 Foods containing contaminated raw vegetables have been implicated in many shigellosis
outbreaks.
 Other foods that are common vehicles for the spread of shigellae include tossed salads,
potato salads, chicken, and shellfish.
 The survivability of Shigella in packaged vegetables (sterile and unsterile) held at
different temperatures (5 °C, 10 °C, and room temperature) was reported.
 In addition, Shigella can survive at room temperature for up to 50 days in foods such as
milk, flour, eggs, clams, shrimp, and oysters, and only 5–10 days in acidic foods (e.g.,
orange juice, tomato juice, carbonated soft drinks) and 1–2 weeks in refrigerated,
fermented milk.

Food at Risk:

Foods that have been identified in Shigella outbreaks include salads (potato, shrimp, tuna,
chicken, turkey, macaroni, fruit, and lettuce), chopped turkey, rice balls, beans, pudding,
strawberries, spinach, raw oysters, luncheon meat, and milk.

Characteristics and Growth Conditions:

 Shigella, (genus Shigella), genus of rod-shaped bacteria in the family Enterobacteriaceae,

species of which are normal inhabitants of the human intestinal tract and can cause
bacillary dysentery, or shigellosis.
 Shigella are microbiologically characterized as gram-negative, non-spore-forming,
nonmotile bacteria.
 Their cells are 0.4 to 0.6 μm (1 micrometre; 1 μm = 0.000039 inch) across by 1 to 3 μm
long.
 The temperature range for growth of Shigella spp. is 6–8 to 45–47°C (ICMSF 1996).
 Rapid inactivation occurs at temperatures around 65°C.
 In contrast, under frozen (-20°C) or refrigerated (4°C) conditions Shigella spp.

Transmission:

 Shigella species are transmitted by the fecal-oral route, and most infections are
transmitted from person to person
 Shigella infections also may be acquired from eating contaminated food.
Symptoms:

 Diarrhea that can be bloody

 Fever
 Stomach pain
 Feeling the need to pass stool (poop) even when the bowels are empty

Doses:

The infectious dose is as low as 10–200 organisms.

Control System:

 Handwashing! Thoroughly clean your hands using soap and water or hand sanitizer. Do
this frequently, and before eating, drinking, or touching your face. Make sure to clean under
your nails.
 Children should be supervised when washing their hands in day care centers and at home.

 When possible, young children with a Shigella infection who are still in diapers should not
come into contact with uninfected children.

Diagnosis:

 Food samples were processed according to international norms for Shigella ISO 21567:
2004 (International Organization for Standardization, 2004).
 To 25 g of each sample in Stomacher bag, 225 ml of Shigella broth (HiMedia
Laboratories, USA) with the addition of novobiocin (0·5 μg ml−1) were added, and
following homogenization for 1 min in a Stomacher blender, the homogenate was
enriched for 24 h at 41·5°C in an anaerobic environment.
 Multiple plating media with differing selectivity can be used to increase the chances of
Shigella spp. isolation like Mac Conkey agar (MAC) such as the low selectivity medium,
Xylose Lysine Desoxycholate agar (XLD) as intermediate selectivity medium and a
Salmonella-Shigella agar (SSA) and Hektoen Enteric agar (HEA) as a highly selective
media.
 After overnight incubation, Shigella broth was further subcultured on Mac Conkey agar,
Xylose Lysine Desoxycholate agar (Difco, USA) and Salmonella-Shigella agar
(Pronadisa, Spain), and plates were incubated at 37°C for 18–24 h.

Main Disease:

 Shigella bacteria cause an infection called shigellosis.

 Most people with Shigella infection have diarrhea (sometimes bloody), fever, and
stomach cramps. Symptoms usually begin 1–2 days after infection and last 7 days.

Vibrio cholerae
Sources:

Vibrio cholerae, the bacterium that causes cholera, is usually found in food or water
contaminated by feces from a person with the infection. Common sources include:

 Municipal water supplies

 Ice made from municipal water
 Foods and drinks sold by street vendors
 Vegetables grown with water containing human wastes
 Raw or undercooked fish and seafood caught in waters polluted with sewage.

Food at Risk:

 Fruits (except sour fruits),

 Poultry,
 Meat, and
 Dairy products

Characteristics and Growth Conditions:

 Vibrios are microbiologically characterized as gram-negative, highly motile, facultative

anaerobes (not requiring oxygen), with one to three whiplike flagella at one end.
 Their cells are curved rods 0.5 μm (micrometre; 1 μm = 10-6 metre) across and 1.5 to 3.0
μm long, single or strung together in S-shapes or spirals.
 Vibrios are able to grow over a wide temperature range (20°C to >40°C) and tend to
grow best under alkaline conditions, although most species of Vibrio will grow between
pH 6.5 and 9.0.

Transmission:

 A person can get cholera by drinking water or eating food contaminated with cholera
bacteria.
 In an epidemic, the source of the contamination is usually the feces of an infected person
that contaminates water or food.
 The disease can spread rapidly in areas with inadequate treatment of sewage and
drinking water.

Symptoms:

 Profuse watery diarrhea, sometimes described as “rice-water stools”

 Vomiting.
 Thirst.
 Leg cramps.
 Restlessness or irritability.

Doses:
  If V cholerae is ingested with water, the infectious dose is 10¿ 9organisms.
 If V cholerae is ingested with food, the infectious dose is 10^3 organisms

Control System:

 Make sure to drink and use safe water to brush your teeth, wash and prepare food, and
make ice
 Wash your hands often with soap and safe water
 Cook food well (especially seafood), keep it covered, and eat it hot. Peel fruits and
vegetables

 Avoid raw foods other than fruits and vegetables that you have peeled yourself
 Clean up safely in the kitchen and in places where the family bathes and washes clothes

Diagnosis:

Enrichment Method

For enrichment method, 10 g of sample was weighed into a sterile stomacher bag and added with
90 mL of alkaline peptone water. The mixture was homogenized in the stomacher for 1 min at
250 rpm and incubated at 37°C for 24 h.
Isolation of Vibrio cholerae
A loopful of the incubated samples was taken from the top pellicle and streaked onto duplicate
selective medium TCBS agar plates. The agar plates were then incubated for 18−24 h at 37°C.
The presumptive colonies of V. cholerae (yellow colonies with 2-3 mm diameter) on TCBS agar
plate.

Rapid tests and other tools

Antigen detection − Several rapid antigen detection-based tests are commercially available for diagnosis
of cholera. These include immunochromatographic lateral flow devices (dipsticks), such as Crystal VC,
which detect the presence of the O1 or O139 antigen in watery diarrheal stools, and Cholkit, which
detects only the O1 antigen.

Molecular tests − Molecular testing for V. cholerae (eg, PCR), including tests that use dried fecal spots,
is also feasible; however, thus far practical use of molecular tests has been limited to epidemiologic
research and surveillance
Darkfield microscopy − Darkfield microscopy of rice-water stools (at 400x magnification) can also be
used to evaluate for the presence of highly motile Vibrios, whose shooting star-like motion can be
inhibited by the subsequent addition of specific antibodies. Darkfield microscopy is quite specific for V.
cholerae but lacks sufficient sensitivity to be used reliably for diagnosis.

Main Disease:

Cholera is an acute diarrheal illness caused by infection of the intestine with Vibrio cholerae bacteria.
Vibrio parahaemolyticus

Sources:

 Raw or undercooked shellfish

 Contaminated water
 Sashimi and sushi

Characteristics and Growth Conditions:

V. parahaemolyticus is a rod-shaped or curved Gram-negative bacteria, 0.5 to 1 µm in diameter,

halophilic (grows in 0.5 to 10% NaCl), oxydase positive, sucrose negative and facultatively
anaerobic.
V. parahaemolyticus will grow under aerobic conditions between 18°C and 37°C, but does so
best at 30°C.

Transmission:

 Faecal-oral,
 Food-borne and
 Water-borne

Symptoms:

Vibrio parahaemolyticus infection causes symptoms of gastro including:

 Watery diarrhoea (occasionally bloody diarrhoea)

 Abdominal cramps
 Nausea
 Vomiting
 Fever
 Headache.

Doses:

An infectious dose of V. parahaemolyticus is considered to be 1,000,000 colony-forming units

(CFUs) or more.

Control System:

 Avoid consumption of raw or undercooked seafood such as oysters, especially during warm
summer periods or if at increased risk of severe diseases (such as people with immune
suppression).
 Keep raw seafood separated from ready to eat foods when preparing or storing food.
 Wash hands with soap and clean water before and after handling raw seafood.
 Wash hands after going to the toilet, after changing a nappy and after handling rubbish.
 Avoid using seawater for cooking.
 Avoid exposing open wounds to seawater. If wounds are exposed then wash with soap and
clean water.

Diagnosis:

 All samples were analyzed following the protocols of the International Organization for
Standardization (ISO/TS 21872-1) (Anonymous, 2007).
The samples (10 g) will homogenize in 90 ml alkaline saline peptone water (ASPW) in a
sterile polythene stomacher bag for 1 min.
 Incubation of the first enrichment will be done at 41.5°C ± 1°C for 6 h ± 1 h, after which,
one milliliter volume of the first enrichment culture (taken from the surface
of the broth) will transfer to 9 ml ASPW as the second enrichment broth. Subsequently, a
loop (1μl) of the second enriched broth will streak onto TCBS and CHROM agar TM
Vibrio (CV) agar plates.
The presence of typical colonies of presumptive V. parahaemolyticus will examine after
24 h ± 3 h incubation of the plates at 37°C.
 Typical colonies of V. parahaemolyticus are green, 2-3 mm and purple, 2-3 mm on TCBS
and CV, respectively. Typical colonies of presumptive V. parahaemolyticus will sub
cultured onto Saline Nutrient Agar (SNA), and tested for presumptive identification
(oxidase test, Gram staining, morphology, and motility).

The colonies suspected as V. parahaemolyticus were confirmed by PCR assay.

Main Disease:

Vibrio parahaemolyticus infection can cause various types of illness including gastroenteritis,
wound infections, and, in more severe cases, sepsis.