Tissue cyst-forming coccidia

PARASITES PROTOZOA

Neospora

• 2 main species
  • Neospora caninum in the dog
  • Neospora hughesi in the horse

• Sporulated oocysts measuring just 10μm

• Oocyst contains 2 sporocysts with 4 sporozoites

• Route of transmission not fully understood

• Often misdiagnosed as Toxoplasma gondii
  • Sarcocystis cysts have thicker walls

• Infection diagnosed by IFAT, ELISA or PCR
  • Identification of lesions and organisms in tissue using immunohistochemical staining
  • Eliminate other causes of abortion first

Life cycle

• Life cycle similar to Toxoplasma gondii

• Limited range of warm-blooded intermediate hosts
  • Asexual reproduction occurs in intermediate host forming tissue cysts

• Host range of sexual stage is unknown for N.caninum
  • Intermediate host for N.hughesi is the horse, but the definitive host is unknown

• Final host
  • Dogs pass oocysts
  • Role not fully understood in pathogenesis
  • 5 day prepatent period
  • Other wild canids may also act as final definitive hosts

• Intermediate host
  • Mostly cattle
  • Natural infection has been documented in other herbivores

• Transmission
  • Transplacental infection occurs in all intermediate hosts and in the canine final host
    • Transplacental can occur in successive pregnancies
  • In cattle, vertical transmission occurs
    • Post-natal infection occurs but is less common

Pathogenesis

• Dogs
  • Occurs mainly in puppies
  • Causes ascending paralysis, especially of hind limbs, with muscle wasting
  • Causes sudden collapse due to myocarditis
  • More than puppy in a litter may be affected, although this may not occur simultaneously
  • Successive litters affected

• Cattle
  • Commenest cause of infectious abortion in dairy cattle
  • Congenitally infected calves can have encephalomyelitis and paresis
  • Abortion usually occurs between 5-7 months of gestation but can occur as early as 3 months
  • No other clinical signs in the cow
  • Repeat abortions possible in same cow (persistently infected)

• Horses
  • Myeloencephalitis
  • Transplacental infection occurs
  • Disease only diagnosed in USA

• Neospora caninum in myositis

Prevention and Control

• Do not allow dogs access to calving cows, placental membranes and aborted or dead calves

• Do not allow dogs to defecate in cattle feeding areas

• Identify and cull seropositive cattle, or do not breed from them or their progeny

• Select seronegative cattle for breeding

• Vaccinate
  • Only in the USA
  • Neoguard or Intervet
  • Killed protozoal vaccine for healthy, preganant cows
  • Dosed in first 3 weeks of pregnancy and then every 3-4 weeks during gestation
  • Revaccination with 2 doses during each subsequent pregnancy

Sarcocystis

Sarcocytis Life Cycle Diagram - Dennis Jacobs & Mark Fox RVC

Sarcocytis - Joaquim Castellà Veterinary Parasitology Universitat Autònoma de Barcelona

• Most infections are asymptomatic

• Heavy infections are causes of chronic wasting in large animals, hide sondemnation and downgrading of carcasses

• Sarcocystis should be differentiated from other tissue-cyst forming coccidia

• There are many species of Sarcocystis which differ in size from microscopic to several centimetres in length
  • S.neurona is an important equine pathogen in the USA

• Infective cyst in the intermediate host is called a sarcocyst

Life Cycle

• Individual life cycles incompletely misunderstood

• Indirect life cycle

• Life cycle alternates between the final and the obligatory intermediate host

• Only one final and one intermediate host

• Sporulated oocyst has 2 sporocysts containing 4 sporozoites
  • Naked oocyst usually seen in faeces as the oocyst wall is very delicate
  • Oocyst measures 15μm in length

• No schizogony in final host

• Gametogeny occurs deep in subepithelial tissue

• Faecal oocyst count is low

• Oocysts are sporulated when passed
  • Difficult to find on faecal examination as the sporocysts are few in number and small

• Ingestion of sporocyst by intermediate host
  • 2 phases of rapid asexual reproduction in vascular endothelial cells
  • Slow multiplication of bradyzoites in muscle tissue
  • Sarcocyst forms with bradyzoites inside, surrounded by a cyst wall and divided into compartments

Epidemiology

• Final hosts are carnivores and omnivores

• Intermediate hosts are herbivores and omnivores

• Humans are the final host for some species and the intermediate hosts for others
  • Final host for species infecting cattle and pigs

• Dogs are final hosts for species infecting cattle, sheep, goats, pigs and horses

• Cats are final hosts for species infecting cattle, sheep and pigs

Pathogenesis

• Widespread infection but mostly asymptomatic

• Cause meat inspection losses

• Sarcocystis in myositis

• Experimental infections cause severe, acute pyrrhexic disease when the organism multiplies in the vascular endothelium

• Can cause chronic wasting disease in cattle and horses
  • Causes abortion and post-natal disease in sheep

• Equine protozoal myeloencephalitis
  • Necrotising encephalomyelitis affecting the grey and white matter of the CNS
  • Caused by S.neurona
  • Opossum thought to be the definitive host
  • Horses thought to be accidental hosts
  • Natural intermediate hosts currently unknown
  • Western Blotting shows 50% of horses in the USA are seropositive
  • Risk factors poorly understood
  • Causes spinal cord dysfunction
    • Ataxia and paralysis

Toxoplasma

Toxoplasma gondii - Ke Hu and John Murray

Toxoplasma Life Cycle Diagram - Dennis Jacobs & Mark Fox RVC

• 1 major pathogenic species called Toxoplasma gondii

• Causes disease in a wide range of animal species including humans

• Important cause of abortion in sheep

• Zoonotic
  • Can cause abortion
  • Can cause congenitally aquired defects

• Forms a sporulated oocyst which is only 10μm
  • Contains 2 sporocysts with 4 sporozoites

• Transmission through ingesting the intermediate host or via the faecal-oral route

Life Cycle

• Complex

• Usually indirect
  • Reffered to as facultatively heterxenous
  • Intermediate host is not essential for completion of the life cycle

• Gametogeny (sexual stage) is host specific for felids

• Any warm blooded animal can act as a facultative intermediate host
  • Asexual reproduction occurs in the intermediate host forming tissue cysts
  • Intermediate host swallows sporulated oocysts or tissue cysts
  • Can be transferred between intermediate hosts by carnivorism

• Cats
  • Sporulation occurs in 2-3 days
  • Cats either swallow infective (sporulated) oocysts where Toxoplasma gondii as a prepatent period of 3 weeks
  • Or eat the tissues of an infective intermediate host where Toxoplasma gondii as a prepatent period of 3-10 days
  • Self-limiting infection
  • Oocysts shed for 1-2 weeks
    • Shedding can occur later if immunity wanes or cat is immunocompromised

• Intermediate host
  • 3 sources of infection
    • Oocysts from environment contaminated by cat faeces
    • Eating cysts in tissues or other infected hosts through carnivorism or undercooked meat
    • Transplacental transmission in some host species during the acute phase of infection

• Acute phase of infection
  • After infection of the intermediate host the organism undergoes a phase of rapid division and dissemination throughout the body
    • Parasite enters cell and asexual reproduction occurs by endogeny (budding) producing 8-16 tachyzoites
    • Tachyzoites are released when host cell bursts
    • Haematogenous spread as more cells are infected
    • Infection continues until the animal develops an immunity in 2 weeks when the infection enters the chronic phase

• Chronic phase of infection
  • Occurs once the host's immune response has become effective
    • Groups of slow growing intracellular bradyzoites become walled off forming infective cysts
    • Bradyzoites inside cysts are protected from the host immune response whereas extracellular tachyzoites are killed
    • Cysts remain viable for months to years and are particulary numerous in muscle and nervous tissue
    • If immunity is suppressed the infection can revert to the acute form

Pathogenesis

• Cat
  • In the intestinal phase of infection only the superficial cells at the tips of the villi are affected
  • Little significant pathogenicity

• Sheep
  • Mostly asymptomatic
  • If a non-immune ewe is infected during pregnancy the consequences will be serious
    • Infection during the first trimester leads to resorbtion
  • Infection during the second trimester leads to fetal death and mummification
  • Infection during the last trimester leads to a weak or stillborn lamb
  • Aborted ewes show focal necrotic placentitis with white lesions in the cotyledons and foetal tissue
  • Diagnosis is confirmed by Giemsa and serology of the ewe's blood

• Clinical outbreaks of toxoplasmosis are sporadic
  • Immunity is aquired before tupping
  • Significant ill-effects are unlikely if immune ewes are infected during pregnancy
  • Not shed from sheep to sheep so predicting outbreaks is difficult

• Humans
  • Mostly asymptomatic
  • Virulent strains cause flu-like symptoms, malaise and/or lymphadenopathy
  • In immunodeficient patients, disease can be caused by even avirulent strains
  • If a non-immune women is infected during pregnancy, abortion or the birth of a congenitally infected child can result
    • E.g. Hydrocephalus, opthalmitis, mental retardation

• Dogs
  • Complication of canine distemper
  • Causes pneumonia and encephalitis

• Cattle and horses
  • Sometimes infectious causing opthalmitis

• Toxoplasma can cause acute interstitial pancreatitis in systemic toxoplasmosis

• Toxoplasma gondii causes myositis

Epidemiology

• Serology
  • Sabin-Feldman Dye test (old method)
  • ELISA
  • Mouse innoculation for confirmation

• Cat
  • 30-80% test seropositive
  • Each cat sheds oocysts for 1-2 weeks of its life

• Human
  • 30% seropositive in UK, 70% seropositive in France

• Meat animals
  • Significant proportion of cattle, sheep, pigs and rabbits can tissue cysts

Prevention

• Cat
  • Impossible if cat is allowed outdoors (will hunt!)
  • If kept indoors, only canned food should be fed and vermin controlled
  • ELISA to check if seropositive

• Human
  • Avoid oocyst ingestion
    • Wash potentially contaminated raw food thoroughly
    • Wash hands after gardening or handling cats and especially before eating
    • Clean out cat litter trays every day before oocysts sporulate
  • Avoid ingestion of tissue cysts
    • Do not eat undercooked meat
    • Wash hands after eating raw meat
    • Take care when lambing or dealing with sheep abortions and stillbirths
    • Pregnant women should avoid lambing altogether when pregnant

• Sheep
  • Toxovax vaccine
    • Live, avirulent strain of Toxoplasma
    • Does not form bradyzoites or tissue cysts
    • Killed by host immune system
    • Single dose given 6 weeks before tupping
    • Protects for 2 years
    • Immunity boosted by natural challenge
  • Medicated feed can be given daily during the main risk period
    • 14 weeks before lambing
  • The best method of protection is to prevent cats from contaminating the pasture, lambing sheds and feed stores