Prevalence and molecular detection of Eimeria species in different types of poultry in Greece and associated risk factors.

Coccidiosis is one of the most notable diseases in chickens having a high economic impact on the poultry industry worldwide. The present study is the first attempt to epidemiologically investigate Eimeria spp. distribution and associated risk factors under different housing and production systems in three major regions in Greece. Faecal samples were obtained from 42 operations (broilers, floor housed, free range and organic layers, backyard farms). A questionnaire was obtained from included operations to acquire additional information regarding farm management, location, production rate and diseases history. Positivity level was 85.7%. All seven Eimeria species were identified, and the most prevalent ones were E. acervulina (79.3%) and E. tenella (65.5%). Single-species and mixed infections were detected in 20.7% and 79.3% of the flocks, respectively. Flock size, type of outdoor area, production system and presence of respiratory disease proved significant risk factors. Flock size up to 10,000 birds correlated strongly (p = 0.02) with higher E. tenella quantities. A very strong correlation (p < 0.001) was found between the presence of respiratory disease and the average OPG level in broiler farms. Organic flocks showed higher prevalence of E. tenella (p = 0.023), while presence of vegetation at the outdoor area correlated strongly with E. brunetti (p < 0.001). Molecular analysis and correlation results in this survey give strong indications although more studies are needed to further understand the involvement of different Eimeria species in various husbandry, production and management systems, to gain more knowledge about the sustainable control of coccidia in poultry.

[Update on toxoplasmosis in poultry farming]. / Update zur Toxoplasmose im Nutzgeflügel.

INTRODUCTION: Toxoplasmosis is a worldwide occurring zoonosis caused by the obligate intracellular parasite Toxoplasma gondii (T. gondii). All warm-blooded species, including humans, serve as intermediate hosts. Definitive hosts are exclusively cats. Farm poultry can become infected with oocysts from contaminated feed or directly from the ground, or by pecking of e.g. infected rodents. Outdoor or free-range housing of poultry increases the risk of infection with length of time. Poulty meat must be seen as a potential source of infection for humans with the increasing popularity of humane animal husbandry practices in poultry farming. This short literature review attempts to assess the current epidemiological situation in farmed poultry and to assess the possible relevance of toxoplasmosis of poultry meat and poultry meat products for human consumption.

INTRODUCTION: La toxoplasmose est une zoonose mondiale causée par le parasite intracellulaire obligatoire Toxoplasma gondii (T. gondii). Toutes les espèces animales à sang chaud, y compris les humains, servent d'hôtes intermédiaires. Les hôtes définitifs sont exclusivement les chats. Les volailles d'élevage peuvent être infectées soit par des oocystes provenant d'aliments contaminés soit directement du sol soit en picorant, par ex. des rongeurs infectés. L'élevage de volailles en plein air ou en parcours augmente le risque d'infection avec le temps. La viande de volaille doit être considérée comme une source potentielle d'infection pour les humains avec la popularité croissante des pratiques d'élevage respectueuses des animaux dans l'élevage de volailles. Cette brève revue de la littérature tente d'évaluer la situation épidémiologique actuelle chez les volailles d'élevage et d'évaluer la pertinence possible de la toxoplasmose de la viande de volaille et des produits à base de viande de volaille pour la consommation humaine.

Bovine Cryptosporidium parvum field isolates differ in cytopathogenicity in HCT-8 monolayers.

Suckling calves are prone to Cryptosporidium infection. The variable degree of clinical disease is influenced by keeping conditions and immune status of the host, but diversity of isolate virulence may also contribute. The aim of the current study was to evaluate the cytopathogenic effects of 26 C. parvum field isolates by using a MTT assay in HCT-8 cell monolayers. Cell viability of monolayers inoculated with oocysts of the field isolates varied considerably with values of 17.7% (± 5.1%) to 99.5% (± 7.1%). A standard deviation of 18.6% was detected for cell viability of the in house reference strain, which were tested alongside in every assay. Field isolates were grouped in three categories of cytopathogenicity. Probably the length of storage has an effect on the level of the cell destruction category detected post infection in vitro. The applied tool may help to better understand the variable course of cryptosporidiosis in the field.

Host-pathogen interaction in Toxoplasma gondii-infected mixed chicken blood cell cultures.

Toxoplasma gondii has the ability to infect various nucleated cell types in different hosts. The aim of the present study was to investigate which chicken blood cells were targeted by T. gondii in a mixed blood cell culture similar to in vivo conditions and to evaluate parasite-host cell interactions. The study consisted of two subsequent experiments. In experiment 1, we applied T. gondii tachyzoites (ME49) at a multiplicity of infection of 1 tachyzoite per blood cell and examined parasite replication, cytokine, and inducible nitric oxide synthase (iNOS) mRNA expression between 1 h and 48 h post-infection (p.i.) by quantitative PCR. By using T. gondii RH-GFP tachyzoites expressing the green fluorescent protein (GFP) in experiment 2, we aimed for visualizing infected cells by confocal laser scanning microscopy (CLSM) and flow cytometric analysis at 24 h p.i. The parasite replication curve showed a massive decrease of parasite stages until 24 h p.i. followed by an approximately plateau phase. We observed mainly significantly increased iNOS mRNA expression levels in T. gondii-infected culture compared to uninfected cells. Flow cytometry and CLSM data confirmed monocytes/macrophages as main target cells for T. gondii. Moreover, different lymphocytes like B cells and cytotoxic T cells seem to be targeted to a low extent. Our findings indicate that monocytes/macrophages play a key role during T. gondii infection in chicken as host cells and triggering of immune response. To the best of our knowledge, this is the first report of a mixed chicken blood cell culture experimentally infected with T. gondii.

Distribution of Cryptosporidium parvum gp60 subtypes in calf herds of Saxony, Germany.

Cryptosporidiosis is a common protozoan parasitic infection that causes diarrhoea in neonatal calves. The high shedding of environmentally resistant oocysts facilitates outbreaks of cryptosporidiosis in humans. In total, 58 farms (512 calves) in Germany (Saxony and Brandenburg) were visited three times each. Faecal samples of pre-weaned calves were microscopically examined for oocysts of Cryptosporidium spp. using Heine staining and were scored with regard to their consistency. Overall, 88.9% of calves tested microscopically positive for Cryptosporidium spp. in at least one sample, and the excretion of oocysts was significantly (P < 0.01) associated with a higher faecal score (diarrhoea). After DNA extraction from pooled farm isolates, 47 samples were successfully subtyped by sequence analysis of the 60 kDa glycoprotein gene (gp60). All isolates belonged to subtype family IIa. IIaA15G2R1 was the most common subtype (present on 66% of the farms), followed by IIaA16G3R1 (13%). Subtypes IIaA14G1R1, IIaA14G2R1, IIaA1612R1, IIaA16G2R1, IIaA17G1R1, IIaA17G2R1, IIaA17G4R1 and IIaA19G2R1 were found sporadically. This is the first description of gp60 subtype IIaA17G4R1 in cattle in Germany.

Experimental Toxoplasma gondii and Eimeria tenella co-infection in chickens.

The widespread apicomplexan parasites Toxoplasma gondii (T. gondii) and Eimeria tenella (E. tenella) are important pathogens with high prevalence in poultry. The aim of our study was the investigation of mutual influences in co-infected chickens, focusing on immune response and course of infection. Two separate trials were performed using in total 96 1-day-old chickens, divided into four study groups: group NC (negative control, uninfected), group PC-T (oral or intramuscular infection with T. gondii oocysts (trial 1) or tachyzoites (trial 2), respectively), group PC-E (oral infection with E. tenella (trial 1) or E. tenella and Eimeria acervulina (trial 2)), and group TE (co-infection). T. gondii and Eimeria infections were validated by different parameters, and cytokine expression in the gut and spleen was investigated. T. gondii-specific antibodies were detected earliest 4 days post infection (p.i.) by immunoblot and direct DNA detection was possible in 22.1% of all tissue samples from infected chickens. Eimeria spp. merogony seemed to be enhanced by co-infection with T. gondii, interestingly without marked differences in oocyst excretion between co-infected and Eimeria spp. mono-infected chickens. An increase of messenger RNA (mRNA) expression of Th1- (IFN-γ, IL-12, TNF-α) and Th2-related cytokines (IL-10) mainly in groups PC-E and TE was observed, however, without statistically significant differences between co-infection and single infection with Eimeria. In conclusion, most of the measurable immune response could be attributed to Eimeria infection. To the best of our knowledge, this is the first report on co-infection experiments of T. gondii with Eimeria spp. in chickens.

Co-infection of Trypanosoma pestanai and Anaplasma phagocytophilum in a dog from Germany.

Infections with arthropod-borne pathogens are an increasing threat world-wide that requires heightened vigilance from veterinary and medical practitioners, especially when they involve new or unusual organisms. A dog was presented to a local veterinary clinic in Germany with malaise, pale mucous membranes and stiff joints. Clinical assessment revealed pyrexia, leukopenia and thrombocytopenia. On suspicion of a tick-borne infection, blood samples were examined for clinical and biochemical parameters and subjected to a Anaplasma phagocytophilum-, Borrelia spp.- and Ehrlichia canis-specific real-time PCR. Additionally, a sample of the pre-therapeutic buffy coat was co-cultured with the Ixodes scapularis cell-line ISE6 for 20days. Only the PCR specific for A. phagocytophilum DNA yielded a positive result, and furthermore, Anaplasma morulae were visible in granulocytes and tick cells. After co-culturing, extracellular trypomastigote and epimastigote stages of Trypanosoma sp. with an average length of 29.7µm were observed, featuring a pointed posterior end. Sequence analysis of a 2080bp fragment of the 18S rRNA gene showed 99% identity to the 18S rRNA gene of Trypanosoma pestanai, previously described from a European badger (Meles meles) in France. The dog's condition improved rapidly in response to doxycycline treatment for three weeks. The clinical status normalized and clinical blood parameters were found to be within the reference ranges. To our knowledge this is the first description of T. pestanai infection in a dog, the first detection of T. pestanai in Germany and the first documented co-infection with these two pathogens. Co-infections with unusual opportunistic vector-borne pathogens should be considered, if acute canine granulocytic anaplasmosis is evident.

[Dirofilaria repens-infection in a dog in central Germany without any history of travel]. / Dirofilaria-repens-Befall bei einem Hund aus Mitteldeutschland ohne Reisevorbericht. Ein Fallbericht.

A mixed breed dog was presented with dyspnoea and fever. In the purulent thorax aspirate, a nematode larva was found during the cytological examination. Subsequent diagnostic tests revealed an infection with Dirofilaria repens, which was probably acquired autochthonously in central Germany. Moxidectin was administered every 4 weeks for 6 months and shown to be effective as indicated by subsequent blood examinations. This case report shows that infection with Dirofilaria repens is possible in Germany and can be treated successfully.

Climate suitability for the transmission of Dirofilaria immitis and D. repens in Germany.

Recently concerns are increasing that dirofilarial nematodes may spread from endemic areas in southern, eastern and central Europe to countries in northern regions of Europe. The increasing number of autochthonous cases of canine Dirofilaria repens infections in Germany indicates that worms of this genus may invade new areas, and climate change may be a key factor in this scenario. Thus analysis of long term development of regional temperature is a pivotal factor in risk analysis related to transmission of these worms. Such information is important for suggestions of counteracting strategies, such as definition of periods of increased transmission risk and, consequently, time slots most suited for preventative measures. In this study, mean daily temperature data from 34 geographical clustered weather stations representing all parts of Germany were analyzed. It is concluded that the increasing trend for average daily temperatures observed in the period from 1984 to 2013 has led to climatic conditions that allow the completion of dirofilarial life cycles in large parts of Germany between May and October. Autochthonous infection with D. repens is already established in some regions and targeted diagnosis and medical prophylaxis is advisable for dogs assumedly exposed during risk of transmission periods. It appears likely that global warming will support further spread of D. repens. Furthermore for the population of dogs the spread of the more pathogenic species D. immitis in hitherto non-endemic Germany is a potential risk if mean temperatures rise to a level suitable for parasite development in the abundant vector mosquitoes during the warmer seasons.

Persistence of Toxoplasma gondii tissue stages in poultry over a conventional fattening cycle.

Toxoplasma gondii is a widely spread protozoon in humans, mammals and poultry. Regarding the latter, nothing is known yet about the duration of T. gondii persistence and distribution over a conventional fattening cycle of turkeys and chickens. Twenty-four turkeys and 12 broiler chickens were infected intravenously with 1×10(6) T. gondii tachyzoites (strain NED). Serum antibody levels were determined weekly by ELISA (turkeys) or immunofluorescent antibody test (chickens). Turkeys were slaughtered at 4, 8, 12 and 16 weeks post-infection (p.i.), and chickens 5 or 10 weeks p.i. (n = 6 per group). Sixteen different tissue samples per bird were analysed for T. gondii by PCR. All infected animals showed seroconversion. In turkeys, 15.9% of all samples were tested positive for T.-gondii-DNA. Among the edible tissues (drumstick, thigh, breast muscle, heart, liver and gizzard) 7.8% tested positive. Among poultry slaughtered after different periods of time after infection no significant differences (P>0.05) regarding the number of positive samples were observed. Only 4 out of 192 samples (2.1%) from infected chickens contained detectable T. gondii DNA.The PCR findings suggested that T. gondii may persist in poultry. Particularly in turkey it was shown that edible tissues stay infected for at least 16 weeks p.i. which indicates a potential risk for consumers of undercooked turkey meat whereas chickens appear less susceptible to T. gondii infection.

Necrotic enteritis in chickens: development of a straightforward disease model system.

The interaction between Eimeria species and Clostridium perfringens was investigated in two different necrotic enteritis (NE) models: 120-day-old broilers were used in two separate experiments consisting of six groups (n=10) each. Besides controls, chickens were infected with coccidia on study day (SD) 18 (Eimeria maxima and Eimeria acervulina (experiment 1) or Eimeria tenella and Eimeria brunetti (experiment 2) and/or a NetB toxin positive C perfringens strain (both experiments: SD 14 or SD 22, respectively)). Body weight, feed intake, mortality rate, clinical disease, Eimeria species oocyst excretion and C perfringens counts were recorded. NE and coccidiosis specific lesion scores were assessed (SD 24 and SD 30). In coinfected groups, NE-typical clinical signs occurred. Coccidiosis-specific lesions were most severe in coinfected groups (significant for E tenella, P<0.05). Most pronounced NE lesions occurred in coinfected chickens compared with C perfringens monoinfected groups (experiment 2, C perfringens infections on SD 22: P<0.05). In experiment 2, E tenella antibody levels were (non-significantly) higher in coinfected groups than in Eimeria species monoinfected groups. Thus, infection with E tenella and Eimeria brunetti followed by C perfringens inoculation is regarded as an easy to handle and suitable model for investigations into NE of chickens.

Effects of pH, sodium chloride, and curing salt on the infectivity of Toxoplasma gondii tissue cysts.

Toxoplasma gondii is one of the most common zoonotic parasites in the world. The parasite causes no or mild symptoms in immunocompetent humans. However, a high potential hazard exists for seronegative pregnant women and immunocompromised patients. The consumption of meat containing tissue cysts or oocyst-contaminated vegetables and fruits or the handling of cat feces poses a high risk of infection with T. gondii. It is known that raw minced meat, raw fresh sausages, and locally produced raw meat products are possible causes of T. gondii infection. The infectivity of T. gondii tissue cysts in meat products depends, among other factors, on the pH and the salt concentration. Therefore, the impact of these two factors on the tissue cysts was examined. For this purpose, dissected musculature and brain from experimentally infected mice (donor mice) were placed in a cell culture medium (RPMI 1640). The medium was adjusted to different pH values (pH 5, 6, and 7) with lactic acid and to different salt concentrations (2.0, 2.5, and 3.0%) with sodium chloride (NaCl) or nitrite-enriched curing salt (NCS) for the various tests. After storage at 4°C for different time periods, the materials were fed to bioassay mice. Later, the brains were examined for presence of T. gondii to assess the infectivity. The data show that T. gondii tissue cysts have a high pH tolerance. Cysts were infectious in the muscle for up to 26 days (pH 5). In contrast to their tolerance to pH, cysts were very sensitive to salt. Muscle cysts survived at an NaCl concentration of up to 2.0% only, and for no longer than 8 days. At NaCl concentrations of 2.5 and 3.0%, the cysts lost their infectivity after 1 day. When NCS instead of NaCl was used under the same conditions, T. gondii muscle cysts retained infectivity for only 4 days at 2.0%. Consequently, NCS (NaCl plus 0.5% nitrite) has a stronger effect on T. gondii cysts than does common table salt. Sausages produced with low NaCl concentration and short contact times pose a potential risk for susceptible individuals.

Experimental Toxoplasma gondii oocyst infections in turkeys (Meleagris gallopavo).

Toxoplasma (T.) gondii is a protozoan parasite with a broad range of intermediate hosts. Humans are often infected by ingestion of tissue cysts in raw or undercooked meat or meat products. Turkeys as food-producing animals can also serve as intermediate hosts. The aim of the present study was to investigate occurrence and predilection sites of T. gondii infection in turkeys after oral infection with oocysts. Experimental infections with different doses of T. gondii oocysts were performed in 36 turkeys to mimic natural infection. Systemic distribution of parasitic stages was investigated by screening 14 different tissues including the edible tissues heart, liver, thigh, breast and drumstick muscle. Parasite detection was based on a conventional nested polymerase chain reaction (PCR). Animals were sacrificed 6-12 weeks after infection. Results demonstrated parasite spreading over the whole organism after oral infection by oocysts. Most frequently affected tissues were brain (47.2% of all brains were positive for T. gondii) and thigh muscle (25.0% positive samples). Other muscles were regularly T. gondii-positive, all other sampled tissues were positive at least once. Thus, edible tissues are one of the predilection sites of T. gondii in turkeys which renders raw or undercooked turkey meat a potential risk for parasite transmission to humans. Data were compared to results from previous parenteral turkey infections with tachyzoites. With the exception of brain, liver and breast muscle affection, no significant differences were observed between both infection routes. Both infection models could be used for research purposes with certain advantages and disadvantages.

Tissue tropism of Toxoplasma gondii in turkeys (Meleagris gallopavo) after parenteral infection.

Turkeys are known to be natural hosts for the zoonotic protozoan parasite Toxoplasma gondii. The objective of the present study was to gain further knowledge of possible predilection sites of T. gondii infection in this species after parenteral application of tachyzoites. A total of 38 turkeys were infected with different doses of T. gondii tachyzoites. Birds were killed either 6 to 8 or 10 to 12 weeks after the experimental infection. Fourteen different tissues per bird were investigated by a nested polymerase chain reaction (PCR) for the presence of the parasites' DNA. T. gondii DNA was found in any type of tissue analysed; in 86.1 % of all infected birds, at least one sample was tested positive. Over all intravenously infected birds, 15.4 % of all analysed samples contained T. gondii DNA. Most frequently affected tissues were liver (43.3 % positive samples), breast muscle (26.7 % positive samples) and heart (20.0 % positive samples), while the brain was less frequently positive (6.7 %). The number of positive tissues varied from zero to seven tissues per animal with at least one T. gondii-positive edible tissue sample in 80 % of all intravenously infected birds. Still, the results did not indicate defined target tissues or a cyst distribution pattern. Nonetheless, edible organs were most frequently parasitised. The number of positive findings did not differ between the early and the late examination time points. Therefore, a persistence of the tissue stages until the end of the study (12 weeks after infection) is concluded.

[Effect of distribution of eggs of strongyles and Parascaris equorum in faecal samples of horses on detection with a combined sedimentation-flotation method]. / Einfluss der Verteilung von Strongyliden- und Ascarideneiern in Pferdekotproben auf den Nachweis mit einem kombinierten Sedimentations-Flotations-Verfahren.

OBJECTIVE: Results of parasitological examination of faecal aliquots may vary between diagnostic laboratories. To examine whether inhomogeneous distribution of worm eggs in faecal samples is responsible for this observation, horse faeces provided for routine diagnosis of helminth infection were examined. Distribution of worm eggs was assessed by examining aliquots taken from different locations of the faecal sample by a combined sedimentation-flotation method (KSFV). In addition, it was tested, whether the homogenization of a larger amount (minimum of 40 g) of faeces before performing KSFV improved reproducibility of the method. MATERIAL AND METHODS: 51 faecal samples of horses were examined three times in parallel by KSFV with ZnSO4 solution. 10 g aliquots were taken from the margin (R), from inside (I) and from both locations (G). The remaining amount of faeces was weighed, suspended with water 1:1 and homogenized. Subsequently, three subsamples, each consisting of 20 g of this suspension, were taken and examined by KSFV. RESULTS: The egg numbers of the nematodes (strongyles and Parascaris equorum ) found in samples that originated from different locations were similar and variation was low. The homogenization of a larger amount of faeces had no relevant impact on egg counts of these nematodes. CONCLUSION AND CLINICAL RELEVANCE: Nematode infections are relevant and frequently occurring in the horse, and reliable assessment of worm egg excretion is a critical aspect for rational planning of control measures. It could be shown that the distribution of nematode eggs (strongyles and Parascaris equorum ) in horse faeces is quite even and results are in principle reproducible if 10 g faeces are examined by KSFV. The homogenization of a larger amount of faeces does not improve the sensitivity or reproducibility of KSFV, and is thus dispensable. For diagnostic purposes, it is advisable to ship approximately 50g of horse faeces to the laboratory.

An invasion record for the swimbladder nematode Anguillicoloides crassus in European eel Anguilla anguilla in a deep warm-monomictic [corrected] lake, from invasion to steady state.

This study is the first account of the establishment and development of the neozoic nematode parasite Anguillicoloides crassus in its host, the European eel Anguilla anguilla, in a deep, warm-monomictic [corrected] lake. A 21 year study of A. crassus took place in Upper Lake Constance (ULC), Europe's second largest pre-alpine lake. The study included two extensive surveys, one in 1991 during the initial parasite invasion phase and the second in 2006 when the infection was well established. The subtropical swimbladder nematode A. crassus was first recorded in A. anguilla in ULC in 1989. Prevalence reached 60% in 1992 and remained at this level until 2007. In 2008, prevalence decreased to 48%. Infection intensity peaked in 1993 at a mean value of 16 adult parasites per host fish. Around 90% of all A. anguilla examined displayed swimbladder lesions, with a significant trend to increasing severity over time. Moreover, heavy swimbladder lesions were seen in c. 10% of A. anguilla ready to migrate to their spawning habitat. Both ruffe Gymnocephalus cernuus and sunfish Lepomis gibbosus serve as paratenic hosts for A. crassus in ULC. Gymnocephalus cernuus seems to be the main vector, and infection is especially frequent in spring possibly caused by reduced immune system efficacy of G. cernuus during winter. In 1991, hypochromic anaemia was prevalent in ULC A. anguilla acutely infected with A. crassus, whereas in 2006 blood values were indicative of chronic infection. The growth and survival rates of A. anguilla during their continental phase were not noticeably altered in infected fish, but damage to the swimbladder probably impairs migration potential and thus the subsequent breeding success of the oceanic phase.

Single oocyst infection: a simple method for isolation of Eimeria spp. from the mixed field samples.

The study described a simple method for single oocyst infection which is usually used to maintain the Eimeria spp. as pure strains in the laboratory and to isolate a single species from the mixed field samples.

Occurrence and molecular characterization of Cryptosporidium spp. genotypes in European hedgehogs (Erinaceus europaeus L.) in Germany.

Juvenile hedgehogs having insufficient body weight are often brought for overwintering to hedgehog rehabilitation centres. Faecal samples of juvenile hedgehogs and overwintering hedgehogs (n=188) collected prior to releasing them back into the wilderness were examined for the presence of Cryptosporidium coproantigen and oocysts. Altogether 56 (29.8%) submitted samples were positive for coproantigen. Forty-five (39.5%, n=114) of the positive samples originated from newly rescued hedgehogs, while 11 (14.8%, n=74) positive samples were from animals that spent several months at the station. Fifteen samples subjected to PCR-RFLP analysis on the partial 18S rRNA locus suggested the presence of C. parvum. Multilocus sequence typing on partial 60 kDa glycoprotein gene, 18S rRNA, actin gene, 70 kDa heat shock protein gene sequences revealed 3 different subtype families: IIa, IIc and a new, proposed as VIIa subtype family. Cryptosporidium sp. genotype belonging to VIIa subtype family is closely related to C. parvum but is genetically distinct being probably a hedgehog-specific Cryptosporidium sp. genotype with unknown zoonotical potential. Hedgehogs excreting Cryptosporidium oocysts represent a potential source for human infections, but also an anthroponotic nature of the IIc subtype family should be reviewed.

Combination of cell culture and quantitative PCR (cc-qPCR) to assess disinfectants efficacy on Cryptosporidium oocysts under standardized conditions.

Oocysts of Cryptosporidium parvum are resistant to environmental conditions and many disinfectants. A combination of cell culture and quantitative real time PCR (cc-qPCR) is established for evaluation of anticoccidial disinfectants against C. parvum. C. parvum oocysts were treated with disinfectants, washed and oocysts were incubated with HCT-8 cell monolayers in the presence of excystation medium for 3h. Subsequently, unbound parasites were removed by washing with growing medium and the infected monolayers were further maintained in fresh growing medium for 48h. Genomic DNA was extracted from each sample and qPCR performed targeting a specific sequence of the 70kDa heat shock protein gene in order to quantify development. Treatment of oocysts with cresolic disinfectants demonstrated dose dependent reduction of viability of oocysts. More than 98% inactivations were recorded with at least 2% concentration of cresolic disinfectants after 2h of treatment. Bleach (sodium hypochlorite) at 6% solution induced 92.7% inactivation of C. parvum oocysts after 2h. Thermally treated oocysts (56 and 70 degrees C for 20min) demonstrated complete inactivation, whereas at 38 degrees C no inactivation was observed. Application of Neopredisan((R)) 135-1 and Aldecoc((R)) TGE (4% for 2h) as recommended according to the current guidelines stipulated by DVG (German Veterinary Society) consistently inactivated more than 99.5% of oocysts. The suggested cc-qPCR method appeared to be suited for standardized testing of inactivation measures, particularly for evaluation of chemical disinfectants and thus cc-qPCR is proposed as an alternative to the established chicken infectivity model for Eimeria tenella for testing anticoccidial disinfectants. A minimum inactivation of 99.5% in cc-qPCR model is claimed as a suitable threshold for certification of chemical products for disinfection of coccidia oocysts.