STAPHYLOCOCCOSIS, STREPTOCOCCOSIS & CAMPYLOBACTERIOSIS

DEFINITION

Staphylococcosis is acute systemic or chronic disease of birds characterized most frequently by purulent arthritis and tenosynovitis. OCCURRENCE

• Staphylococcal infections of poultry occur worldwide and affect all classes of birds. • Outbreaks are most important in turkeys and broilers. • The organisms are common in the environment and are especially associated with the skin (normal inhabitant). • Most diseases produced by Staphylococcus sp. are associated with ahistory of break in the skin or beak (trauma, beak trimming, toe trimming, foot pad burns etc.). OCCURRENCE

• Avian infections tend to be caused by types occurring in birds rather than human strains. • Isolates pathogenic for one class of poultry are usually pathogenic for other classes of birds. • Toxigenic strains capable of causing food poisoning can contaminate the skin of processed poultry (zoonotic importance). • The source of these strains at present is in debate. • Bio-typing indicates processing plant worker origin while plasmid profile indicates poultry origin. HISTORICAL INFORMATION

• Staphylococci were first discovered to be a cause of arthritis in geese in 1892. • Since that time they have been identified as the cause of a variety of localized and systemic diseases in many different avian species and in most areas of the world. • The disease was more common in turkeys when they were raised on range than it is now. ETIOLOGY

• 1. Most staphylococci isolates have been identified as Staphylococcus aureus, a Gram-positive coccus occurring in clusters (grapes like). Pathogenic isolates are usually coagulase positive.

2. Organisms are moderately resistant to common disinfectants. • Chlorine-containing disinfectants are efficacious in the absence of organic material. 3. Toxins produced by staphylococci can increase both the virulence and pathogenicity of a particular strain. Clinico-pathological picture

• General signs of illness. • Fever and reduced appetite. • Foot abscess (bumblefoot). • Lameness and arthritis. • Birds unable to move. • Omphalitis in newly hatched chicks. • Reduced fertility and hatchability in breeders. LESIONS Diseases produced by staphylococcus infections include:

1. Omphalitis. 7. Discospondylitis (spondylitis). 2. Gangrenous dermatitis. 8. Osteomyelitis. 3. Cellulitis. 9. Endocarditis. 4. Abscesses (bumble foot). 10. Green liver-osteomyelitis complex in turkey. 5. Septicemia. Sternal bursitis. 6. Arthritis/periarthritis/synovitis. 11. 1. Omphalitis

• Although infections of the yolk sac occur, they are less common than omphalitis caused by other bacteria as (Aerobacter, Proteus, Klebsiella, Pseudomonas, Salmonella, Bacillus, enteric Streptococcus, E. coli, …etc.). • Sources of the bacterium include the breeder flock, hatchery environment, and hatchery workers. 2. Gangrenous dermatitis

• Affected areas of the skin are dark red, moist, crepitation sound, thickened, and clearly demarcated from adjacent normal skin. • Usually traumatic lesions such as punctures or scratches are present. • The serosanguinous fluid seen in clostridial infections is minimal or absent. Staphylococcal gangrenous dermatitis is typically secondary to immunosuppression caused by infectious bursal disease or chicken infectious anemia virus. • Immune suppression (CIA, IBD, MD, Reo)+ Staph + Cl. Septicum ↓ Gangarenous dermatitis (blue wing disease)

3. Cellulitis

• A purulent inflammation is present in subcutaneous tissues. • Traumatic lesions may or may not be present. • The overlying skin tends to be dry and discolored. 4. Abscesses

• There are localized purulent lesions in the skin. • The plantar surface of the foot is a common site and results in bumble foot. • Abscesses result from puncture wounds.

5. Septicemia

• There is an acute increase in mortality with congestion of the internal organs, hyperaemia, enlargement, sub –capsular hemorrahge and various- degree coagulative necrosis in the liver or the spleen are observed. • It is usually associated with a processing event of the bird such as beak trimming or some other trauma to the skin. • As staphylococci are ubiquitous, their presence could not be prevented . • The measures should be directed toward minimizing the possibilities for traumas of skin, respiratory and intestinal mucosa.

6. Arthritis/periarthritis/synovitis

• Any joint, tendon sheath, or synovial bursa can be affected. • Arthritis/periarthritis/synovitis is seen clinically as swollen, hot joints, especially hock joints. • It occurs as a sequel to septicemia and can be experimentally reproduced by intravenous injection of pathogenic strains. • Initially, affected tissues are acutely inflamed and contain white to yellow soft fibrinopurulent exudate. Later, the exudate becomes caseous. • Fibrosis of affected tissues occurs late. • Affected birds often have bile stasis of the liver. • High numbers of large mononuclear cells are seen in blood smears.

7. Discospondylitis (spondylitis)

• The joints of articulating thoracolumbar vertebrae are affected. • The process spreads to affect adjacent vertebrae. • Lesions may become so extensive that pressure on the spinal cord will develop causing paresis and paralysis . 8. Osteomyelitis(fumer head necrosis)

• This is a sequel to septicemia. • Organisms localize in metaphyseal vessels invading the cartilage of the growth plate of actively growing bones. • Initially, pale yellow, friable bone is seen in affected areas adjacent to the growth plate, especially in the proximal tibia and metatarsus. • Necrotic areas, abscesses, and sequestra are seen later.

9. Endocarditis

• This is an uncommon sequel to septicemia. • There are vegetations on the mitral and/or aortic valves. • Emboli from valve lesions cause infarcts in the brain, liver, and spleen. 10. Green liver-osteomyelitis complex in turkey

• This is a condition observed at slaughter. • Normal-appearing processed turkey carcasses present green discoloration of the liver and associated arthritis/synovitis, soft- tissue abscesses, and osteomyelitis of the proximal tibia. • Staphylococcus aureus is most commonly isolated from these lesions but other opportunistic bacteria such as Escherichia coli have also been isolated. Green liver-osteomyelitis complex Other local form of Staphylococcosis is Sternal bursitis.

• The sternal bursa is enlarged at a various extent because of gathering of purulent substance. • Sometimes, the covering skin is necrotized. • The microorganism is widely distributed in the environment and mainly on the skin. • Most of Staphylococcus aureus-induced lesions are associated with skin injuries, debeaking, finger cuts. • All categories of birds are affected.

Diagnosis

1- field diagnosis: • Flock history • Clinical signs • PM lesions 2- Lab. diagnosis. DIAGNOSIS

1. Gross lesions are suggestive. A rapid, presumptive diagnosis can be made by identifying direct smear stained with Gram stain is showing the typical cocci in smears from lesions. 2. Organisms can be readily cultured and identified from lesions and often from the livers of affected birds.  On mannitol salt agar appear as golden yellow colonies.  On blood agar appear as rounded colonies with Beta hemolysis. DIAGNOSIS

• Biochemical identification. • Molecular identification. CONTROL

1. Because staphylococci are ubiquitous in the environment their presence cannot be prevented. When an outbreak is associated with a particular environment, the source should be sought and eliminated. 2. Protect broilers from infectious bursal disease with an appropriate vaccination program. 3. Take measures to reduce the occurrence of traumatic skin lesions and foot pad burns, as well as any enteric disease which would damage the integrity of the intestinal mucosa. CONTROL

4. The respiratory tract has also been identified as an important portal of entry for pathogenic staphylococci in turkeys. Exposing chickens or turkeys to a live avirulent vaccine, namely strain 115 of Staphylococcus epidermidis, by aerosol at 10 days and again at 4-6 weeks substantially reduced the incidence of staphylococcosis and improves overall flock livability. 5. Avoid overly severe feed restriction in breeder replacements which has been associated with an increased incidence of staphylococcosis. TREATMENT

1. Broad spectrum antibiotics are effective against staphylococci if given early in the course of the disease.

2. Resistance to antibiotics is common and isolates should be tested for sensitivity.

3. Usually treatment is not cost effective and preventive programs should be relied on.

4. Treatment: based on sensitivity testing; Penicillin, streptomycin, tetracyclines, erythromycin, novobiocin and sulfa drugs

INTRODUCTION

Streptococcosis in avian species is worldwide in distribution, occurring as both acute septicemic and chronic infections with mortality ranging from 0.5%–50%. HISTORY

• Acute streptococcal infections of poultry were first described in chickens in 1902 and 1908 as a septicemia. • Chronic streptococcosis caused 50% mortality in a flock over a 4- month period and was identified as the cause of mortality due to salpingitis and peritonitis in chickens. • Streptococcosis in turkeys was reported as early as 1932. • Bacterial or vegetative endocarditis associated with streptococci was first reported in 1927 and again in 1947. ETIOLOGY

•The genus Streptococcus is composed of Gram-positive, non-motile, non-spore forming, facultative anaerobes, spherical bacteria occurring in singles, pairs, and short chains. ETIOLOGY oStreptococcus spp., isolated from avian species and associated with disease, includes: S. bovis - causing acute septicemia and joint infections found in racing pigeons. S. dysgalactiae - has been cultured from broilers with cellulitis, a condition observed on the skin and subcutaneous tissue at processing. S. pleomorphus - an obligate anaerobe in normal cecal contents of chickens, turkeys, and ducks. S. mutans a common bacterium in the human oral cavity, has been associated with septicemia and mortality in geese. S. zooepidemicus - resulting in bacterial endocarditis. S. gallinaceous. ETIOLOGY

• S. zooepidemicus infection is very common in mature birds. • Other Streptococcal infections are commonly seen in birds of all ages (especiallyoung birds) and also isolated from contaminated eggs or dead embryos. PATHOBIOLOGY AND EPIDEMIOLOGY

• S. zooepidemicus occurs almost exclusively in mature chickens but has been documented as a cause of mortality in wild birds. • Experimentally, rabbits, mice, turkeys, pigeons, ducks, and geese are susceptible. • Transmission of streptococci occurs most commonly via oral and aerosol routes. • Transmission can occur, however, through skin lesions, especially in caged layers. • Aerosol transmission of S. zooepidemicus results in acute septicemia in chickens. • Incubation periods range from 1 day to several weeks, with 5–21 days being most common. • • MOI: oral, aerosol and may also through injured skin • Endocarditis occurs when septicemic streptococcal infection progresses to a subacute or chronic stage. CLINICAL SIGNS

• S. zooepidemicus infections, clinical signs are typical of an acute septicemic infection and include: yellow droppings and emaciation, blood stained tissue and feathers around the head, pale combs and wattles, cyanosis in the terminal stages also has been described. • Mortality can range from low to 50%. • Breeders or layers may have an egg production drop of as much as 15%. Streptococcus spp. has been isolated in cases of acute fibronopurulent conjunctivitis. • In pigeons, S. bovis infection produced the acute onset of mortality with occasional lameness, in-appetence, diarrhea, and the inability to fly. GROSS PATHOLOGY

Gross lesions of S. zooepidemicus in acute disease are characterized by: • Splenomegaly and hepatomegaly (with or without miliary to 1cm red, tan, or white foci), enlarged kidneys, congestion of subcutaneous tissue, and peritonitis. • Subcutaneous and pericardial fluid may appear serosanguineous. • Blood stained feathers around the mouth and head with blood coming from the mouth. GROSS PATHOLOGY

Gross lesions of chronic streptococcal infections include: • Fibrinous arthritis and/or tenosynovitis, • Osteomyelitis, • Salpingitis, • Fibrinous pericarditis and perihepatitis, necrotic myocarditis, valvular endocarditis (Vegetative valvular lesions are usually yellow, white, or tan; are small; and have raised rough areas on the valvular surface). GROSS PATHOLOGY Strept. zooepidemicus Infection Showing Perihepatitis and Peritonitis GROSS PATHOLOGY

• Enlarged, pale, flaccid heart, pale to hemorrhagic areas in the myocardium, especially at the base of the valves, below the affected valve, or apex of the heart. GROSS PATHOLOGY

• Infarcts in the liver, spleen, or heart and, less commonly, infarcts in the lung, kidney, and brain. • Liver infarctions are usually peripherally located on margins, have a pale creamy color and are sharply demarcated . • The penetration of the infection occurs mainly via the oral or aerogenic route, but could also enter through the injured skin, especially in battery cage layer hens. GROSS PATHOLOGY

• In broilers, cellulitis involving the skin and subcutaneous tissues observed at processing. • In pigeons, congestion of the spleen and liver with accumulations of fluid around the pectoral muscles. DIAGNOSIS

Isolation and Identification of Causative Agent • Demonstration of bacteria typical of streptococci in blood films or impression smears of affected heart valves or lesions from birds with typical signs and lesions provide a presumptive diagnosis of streptococcosis. DIAGNOSIS

• Streptococci are readily isolated on blood agar and appears as hemolytic colonies or on mannitol salt agar appears as pink colonies. DIAGNOSIS

Serology: • A rapid detection test by latex agglutination has been described for the identification of antigenic serogroup of streptococci. Molecularly: • By using PCR. DIFFERENTIAL DIAGNOSIS

Differential diagnosis includes other bacterial septicemic diseases: • Staphylococcosis, • Colibacillosis, • Pasteurellosis, • Erysipelas infection,….etc.,. TREATMENT

• Antibacterial susceptibility should be performed on bacterial isolates in any clinical cases of streptococcosis. • Clinically affected birds respond well early in the course of the disease • As the disease progresses within a flock, treatment efficacy decreases. • Treatment in acute and subacute infections includes the use of antibiotics such as Penicillin, Erythromycin, Novobiocin, Oxytetracycline, Chlortetracycline, Tetracycline. • There is no treatment for poultry with bacterial endocarditis. • In vitro susceptibility of S. bovis from pigeons has been demonstrated to the following antibiotics: Penicillins, Macrolides, Lincomycin, Tetracyclines, Chloramphenicol and Nitrofurans. PREVENTION AND CONTROL

Prevention and control require following: • Reducing stress. • Preventing immune-depressive diseases and conditions. • Proper cleaning and disinfection. • The use of formaldehyde reduces the total count of Streptococcus spp. in hatchers by as much as 85.7%.

INTRODUCTION

◦ Campylobacteriosis is currently only a public health concern and not a poultry health concern.

◦ Campylobacteriosis more recently has been recognized as a leading cause of human foodborne illness and is even more common than Salmonellosis.

◦ There are many sources of campylobacteriosis in humans, but poultry is often considered as a major source.

◦ It occurred as a disease of egg-laying chickens in the 1950 and was known at that time as vibrionic hepatitis and vibrio-like organisms now known as Campylobacter jejuni.

◦ Layers during these episodes showed a drop in egg production with increased mortality. ETIOLOGY

◦ Thermophilic Campylobacter species C. jejuni and C. coli can be isolated from the intestines of poultry. ◦ It is a micro-aerophilic, small curved or spiral Gram-negative rod with a rapidly motility under a phase-contrast microscope. ◦ the optimal growth temperature of thermo-tolerant Campylobacter lies between 37 and 42 °C (very close to the chicken body temperature), they are not able to grow below 30 °C (room temperature). EPIDEMIOLOGY

◦ Campylobacter is a commensal organism in avian hosts. ◦ Affected Species: domestic poultry including chickens, turkeys, ducks, and geese. ◦ Colonization with C. jejuni occurs mostly in the ceca, more specifically in the lower intestine in the mucous layer covering the intestinal crypts. ◦ Broilers are typically shown to be free of Campylobacter at a day of age and detection in a flock is usually possible at 2 to 3 weeks of age. ◦ Maternal immunity is thought to play a protective role in delaying Campylobacter infection. EPIDEMIOLOGY

◦ Interestingly, experimental studies have demonstrated that day-old chicks are susceptible to colonization and Campylobacter has also been isolated from the ovaries and semen of broiler breeders as well as from paper pad liners in hatching box. ◦ Vertical transmission is still being debated. ◦ Horizontal transmission from the environment to poultry houses is the most common mode of transmission of Campylobacter on poultry farms. ◦ Mode of infection through fecal contact to the water source or with flies as vectors. ◦ Flies as vectors might partly explain the seasonality of campylobacteriosis in poultry flocks with an observed higher prevalence in summer time. EPIDEMIOLOGY

◦ The incubation period ranged from 2–5 days.

◦ Once infection has entered the broiler house, its spreads rapidly; within 10-14 days post-infection, more than 90% of the birds in a flock will be positive.

◦ When Campylobacter positive chickens enter the slaughterhouse with large numbers of Campylobacter in the intestines, as well as on the feathers and skin, this inevitably leads to a cross-contamination of the equipment, environment and other processed birds……”ZONOOTIC HAZARD” EPIDEMIOLOGY Potential sources of infection include

Because Campylobacter growth is very sensitive to oxygen and temperature, the organism is usually unable to grow in feed, litter, or water under normal ambient conditions. So, sources of infection are:  Old litter  Untreated drinking water (passive carrier)  Feed and water contaminated by feces  Other farm animals  Wildlife species  Insects-mechanical vectors  Equipment, transport vehicles and farm workers Pathogenesis

◦ Birds become infected with campylobacters via the fecal-oral route.

◦ As enteric organisms, Campylobacter spp. are able to survive the harsh conditions in the acidic proventriculus and ventriculus (gizzard) as well as in the small intestine, and eventually reach the lower intestine, where they establish colonization in cecal and cloacal crypts.

◦ The organism can also be recovered from the small intestine and the ventriculus (gizzard), and infrequently from the liver, spleen, blood, and gall bladder. CLINICAL SIGNS

◦ Chickens are carriers of Campylobacter and typically do not exhibit clinical signs or lesions.

◦ Egg drop and increased mortality in egg-laying chickens was observed with vibrionic hepatitis, watery/mucoid/bloody dropping, weight loss, or even mortality may also observed in some cases.

◦ Experimental infection in an early report, 3-day old chickens inoculated with a high dose of C. jejuni developed diarrhea within 72 hours, which lasted for 10 days and resulted in considerable weight loss as well as a mortality of 32% . LESIONS

◦ No lesions are observed in carrier birds.

◦ Hepatic necrosis consisting of small stellate or star shaped whitish necrotic foci to fairly large diffuse areas has been described in egg-laying chickens with vibrionic hepatitis.

◦ Accumulation of fluid, gas, or excess mucus, distention of intestines including ceca with watery/foamy material may be a common finding.

◦ Blood and mucus in the lumen of small intestine and petechial hemorrhages in the ventricular (gizzard) mucosa of chicks can be seen occasionally.

DIAGNOSIS

• Samples: Bile is a better source of organism than liver.

• Thermophilic Campylobacter spp., grow optimally at 42 °C on artificial media as slow-

growing fastidious organisms, and require a micro-aerobic atmosphere.

• Campylobacter Gram-negative and non-spore forming, cells are S-shaped spirally curved

rods, and possess a single polar flagellum, mediating a characteristic corkscrew-like or

darting motility.

• Although cells may transform to spherical or coccoid forms in response to stress or

deleterious conditions.

DIAGNOSIS Culture-Based Isolation and Detection Methods ◦ Thermophilic campylobacters are fastidious and slow growing, requiring microaerobic atmosphere (containing 5% O2, 10% CO2, 85% N2) and elevated temperature (42°C) for optimal growth under laboratory conditions. Selective medium for culturing C. jejuniand C. coli ◦ Skirrow, Preston, Karmali, Modified charcoal cefoperazone deoxycholate agar (mCCDA), Campy-Line agar (CLA), Campylobacter agar plates (CAP), Campylobacter selective chromogenic medium CASA. Serology ◦ ELISA Molecularly: ◦ By using PCR.

TREATMENT No treatment is currently used.

Intervention Strategies ◦ Prevention of flock colonization by use of biosecurity-based interventions.

◦ Prevention and/or reduction of Campylobacter colonization by non-biosecurity based measures such as vaccination, bacteriocins, feed additives, bacteriophages in order to competitive exclusion.