Intestinal parasites of the endangered European ground squirrel (Spermophilus citellus) populations in Slovakia.

The present pilot research was focused on the detection of intestinal parasites in the ground squirrel populations in various regions of Slovakia. Only a very little information is currently available on the parasitic species composition of the European ground squirrel in Slovakia and across Europe. In the Slovak Republic, there are 70 locations where the ground squirrel populations are present. A total of 600 faecal samples of the European ground squirrels, collected from 36 locations all over Slovakia, were examined by applying the coprological method. The presence of the protozoan coccidian parasite of the Eimeria genus was confirmed in all of the analysed locations. The presence of eggs of four helminths were confirmed: Capillaria spp. (66.6% of locations); the Trichostrongylidae family (42.8% of locations); Hymenolepis spp. (11.9% of locations); and Citellina spp. (7.14% of locations). Dead individuals that were found in the analysed localities were subjected to necropsy and the tissues scraped off their small intestines were stained in order to confirm the presence of parasites. The post-mortem examination of the intestines and the sedimentation of the intestinal contents in a saline solution did not result in the confirmation of the presence of the eggs, adults or the larval stages of parasites. Spermophilus citellus is one of the strictly protected animal species in Slovakia. In recent years, numerous projects aimed at supporting and protecting ground squirrels have been implemented. The present pilot study on intestinal parasites and the subsequent cooperation with environmental activists will contribute to the support and stabilisation of the presence of these animals in our country.

Coproparasitological examinations and molecular determination of Eimeria species in Madura cattle reared on Madura Island, Indonesia.

Madura cattle, which are native to Indonesia and mainly kept on Madura Island, East Java, are expected to contribute to improving the regional meat self-sufficiency. Eimeria spp. are the most pathogenic protozoans among gastrointestinal parasites in livestock but no molecular surveys of Eimeria spp. in Madura cattle have been conducted to date. In this study, a total of 183 fecal samples were collected from Madura cattle and 60 (32.8%) were positive for parasites of protozoans and nematodes by the sugar floatation method. Among the samples with parasites, Eimeria spp. oocysts were detected in 50 samples (27.3%) with an average OPG value of 1686.1. Eimeria spp. were successfully identified to the species level in 26 samples with Eimeria bovis being the most prevalent, followed by E. zuernii and E. aubrunensis. A total of 21 samples showed mixed infection of more than two species of Eimeria. E. bovis and E. zuernii have been recognized as having high virulency and, thus, these parasites are potential sources of severe coccidiosis and the cause of infections in other cattle. Although additional studies are warranted, these results can be helpful for improving the management and productivity of Madura cattle.

The incidence of necrotic enteritis in turkeys is associated with farm, season and faecal Eimeria oocyst counts.

BACKGROUND: Specific studies on the epidemiology of necrotic enteritis in turkeys are absent in the literature. Necrotic enteritis is common in turkeys and a leading cause of use of therapeutic antibiotics. This study describes the incidence of necrotic enteritis in turkey farms, and the association between incidence and bird age, season, faecal oocyst counts, grow-out size and feed mill. RESULTS: Necrotic enteritis was diagnosed post mortem in 20.2 % of 545 grow-outs of commercial female and male B.U.T. 10 turkeys started during the years 2010-2016. 80 % of all cases occurred at four to seven weeks of age. Median (minimum-maximum) age at disease detection was 37 (18-115) days. Turkey age at detection was influenced by season, and varied from 33 days among grow-outs hatched in February to 42 days among those hatched in July-August. The incidence also varied with season, showing peak occurrence among grow-outs hatched during February-March and the lowest incidence in turkeys hatched in July-August. 59 % of all cases were detected in 25 % of the farms. The incidence per farm varied from below 4 to 59 %. A multilevel mixed-effects logistic regression model indicated clear impacts of farm and season on incidence, and border-line impacts of grow-out size and feed mill. Grow-outs diagnosed with necrotic enteritis had higher counts of faecal Eimeria oocysts than grow-outs without a diagnosis. This difference was particularly clear during the high-risk period at five to seven weeks of age. Necrotic enteritis was the cause of treatment with therapeutic antibiotics in 88.2 % of all cases of treatment. CONCLUSIONS: Our data indicate that necrotic enteritis incidence in turkeys can be substantially influenced by risk factors at farm level. The incidence showed two seasonal peaks; a moderate peak in turkeys hatched in October/November and a marked peak in turkeys hatched during February/March. Mitigation measures at the farm may therefore be of particular importance during these months in farms located in the Northern temperate zone. Measures which effectively reduce counts of faecal Eimeria oocyst are likely to be among the more promising actions to take both at the farm and at population level.

DESCRIPTION OF A NEW EIMERIA SPECIES (APICOMPLEXA: EIMERIIDAE) RESPONSIBLE FOR CLINICAL COCCIDIOSIS IN COMMERCIAL CHUKAR PARTRIDGE (ALECTORIS CHUKAR).

Recurrent coccidiosis affecting a commercial chukar partridge (Alectoris chukar) farm in Ontario, Canada was investigated. The responsible pathogenic Eimeria species was isolated for biological characterization. The uniformity of oocyst morphometrics supported that only a single Eimeria sp. was present. Experimental infections with coccidia-free chukars were used to describe exogenous and endogenous developmental stages of the parasite. The prepatent period of the causative Eimeria species was 5 days and patency lasted 11 days; fecundity was 1,573 to 30,057, with the highest fecundity recorded with the lowest challenge dose. Endogenous development was elucidated histologically from samples collected at 8 locations along the intestinal tract at 26 time points throughout prepatency. The parasite had 5 asexual generations before oocyst formation that were located from the mid-jejunum to the mid-rectum and in the ceca. Sporulation of oocysts suspended in potassium dichromate at room temperature (22 C) occurred within 24 hr. Oocysts (n = 50) averaged 21.8 by 18.6 µm and featured a polar granule; sporocysts (n = 50) averaged 10.9 by 7.1 µm and possessed a Stieda body, sub-Stieda body, sporozoite refractile bodies, and sporocyst residuum. Comparisons with described Eimeria spp. infecting partridges suggest that the biological features of this pathogenic species are unique; similarly, sequences from both mitochondrial and nuclear loci support the naming of this new Eimeria species.

Detection and Quantification of Clostridium perfringens and Eimeria spp. in Poultry Dust Using Real-Time PCR Under Experimental and Field Conditions.

Infection of poultry with Eimeria spp., the causative agent of coccidiosis, can predispose birds to necrotic enteritis (NE) caused by netB gene-positive strains of Clostridium perfringens. The detection of Eimeria spp., C. perfringens, and netB were examined in settled dust from broiler flocks under experimental and field conditions. Dust samples were collected from settle plates twice weekly from two experimental flocks inoculated with three species of pathogenic Eimeria in 9-day-old chicks, followed by netB gene-positive C. perfringens 5 days later to produce subclinical and clinical NE. A noninoculated flock was sampled weekly from day 0 and served as a control flock. An additional 227 dust samples from commercial broiler flocks were collected at the end-of-batch (6-7 wk of age; one scraped dust sample per flock). In the NE-subclinical and NE-clinical flocks, high levels of Eimeria spp. and C. perfringens were detected after inoculation followed by a gradual decline over time. In the control flock, C. perfringens and netB were detected at low levels. No significant effect of sampling location was evident on Eimeria spp., C. perfringens, and netB load within poultry houses. These results provide evidence that Eimeria spp., C. perfringens, and netB gene copies can be readily measured in poultry dust samples collected in settle plates and may provide an alternative sampling method for monitoring flock coccidiosis and NE status. Further studies are required to assess the utility for such a test in commercial flocks.

Artículo regular­Detección y cuantificación de Clostridium perfringens y Eimeria spp. en polvo de instalaciones avícolas mediante PCR en tiempo real bajo condiciones experimentales y de campo. La infección de aves comerciales con Eimeria spp., el agente causante de la coccidiosis, puede predisponer a las aves a enteritis necrótica (NE) causada por cepas de Clostridium perfringens positivas a la presencia del gene netB. La detección de Eimeria spp., C. perfringens y del gene netB se examinó en el polvo sedimentado de parvadas de pollos de engorde bajo condiciones experimentales y de campo. Se recolectaron muestras de polvo por sedimentación en placa dos veces por semana de dos parvadas experimentales inoculadas con tres especies de Eimeria patógena en pollitos de nueve días, seguidas de C. perfringens positiva al gene netB cinco días después para producir enteritis necrótica subclínica y clínica. Una parvada no inoculada se muestreó semanalmente desde el día cero y sirvió como parvada control. Se recolectaron 227 muestras adicionales de polvo de parvadas de pollos de engorde comerciales al final del lote (6 a 7 semanas de edad; una muestra de polvo por raspado por parvada). En las parvadas con enteritis necrótica subclínica y clínica, se detectaron niveles altos de Eimeria spp. y de C. perfringens después de la inoculación seguida de una disminución gradual con el tiempo. En la parvada control, C. perfringens y el gene netB se detectaron en niveles bajos. No fue evidente ningún efecto significativo de la ubicación del muestreo sobre la carga de Eimeria spp., C. perfringens y del gene netB dentro de las casetas. Estos resultados proporcionan evidencia de que las copias genéticas de Eimeria spp., C. perfringens y del gene y netB se pueden medir fácilmente en muestras de polvo de instalaciones avícolas recolectadas mediante sedimentación en placa y pueden proporcionar un método de muestreo alternativo para monitorear coccidiosis y el estado de enteritis necrótica en la parvada. Se requieren más estudios para evaluar la utilidad de tal prueba en parvadas comerciales.

Eimeria spp. in naturally infected beef cattle: Dynamics of oocysts excretion, prevalence, and comparison between parasitological diagnostics.

Eimeria spp. infections cause mortality, reduced well-being, and substantial economic losses implications for cattle production worldwide. The present work followed up the excretion of Eimeria spp. oocysts in two naturally infected beef herds, from two different properties, to investigate the dynamics of oocyst excretion and the prevalence of Eimeria spp. in different animal categories and seasons of the year (rainy season - October to April; dry season - May to September). Even that, the species of Eimeria were identified and the parasitological techniques of Gordon and Whitlock modified and Mini-FLOTAC were used. In both herds, animals up to 14 months had a mean total OPG counts higher than older animals (after 15-16 months of age), and the species E. zuernii and E. bovis were more frequently identified, the first species being more frequent in animals from 1 to 2 months of age, while E. bovis prevailed from three months old. On property 1, the highest mean OPG counts (P ≤ 0.05) were obtained between October 2017 and September 2018, with the highest mean OPG counts in October 2017, when the animals were aged between 4-5 months. The prevalence of the pathogen on property 1 was 59.16 % and 43.62 % in the rainy and dry season, respectively, a higher parasitic load (P ≤ 0.05) was verified in the rainy season. On property 2, the mean OPG counts of Eimeria spp. was higher (P ≤ 0.05) in animals between 8-16 months, with the highest peak in November 2019, when they were one year old. The on-site prevalence during the rainy season on property 2 was 53.09 % and 49.79 % on dry season, and no difference (P = 0.92) in the mean OPG counts of Eimeria spp. during the seasons. There was a difference (P ≤ 0.05) in the count of oocysts in females after 18 months of age than males, which was probably due to the increase in animal density. Both tested techniques can be used for quantification of the excretion of oocysts of Eimeria spp. in cattle feces showing the same OPG mean count (r = 0.9287; p = 0.0025; R² = 0.8625). Mini-FLOTAC showed higher prevalence for Eimeria spp., however, can be an obstacle depending on the number of fecal samples that need to be processed.

A Survey of Coccidia and Nematodes in Pastured Poultry in the State of Georgia.

The aim of this study was to evaluate and quantify the parasitological challenge in pastured poultry production in the state of Georgia. Over the course of 1 yr, fecal samples from six turkey flocks, 10 broiler flocks, and 13 layer flocks were collected on a pastured farm in 2-wk intervals to determine counts of Eimeria oocysts and nematode eggs. Average coccidia counts were 10,198 oocysts per gram of feces (OPG) in broiler flocks, 1470 OPG in layer flocks, and 695 OPG in turkey flocks. The means in broiler and turkey flocks were higher at their first week on pasture. Counts in broilers and layers were significantly higher in spring than in winter and summer. Coccidia counts in broilers were lower than published numbers in conventionally reared poultry, indicating the rotation system of the pastures might effectively reduce the infection pressure. Next-generation sequencing of PCR products showed the presence of most described Eimeria spp. in broilers, layers, and turkeys. In addition, operational taxonomic units (OTUs) x, y, and z were found. The frequency of species was similar for broilers and layers, with the exception that Eimeria praecox and OTU z were more common in layers. In layer flocks, the average count of roundworm eggs per gram of feces (EPG) was 509 EPG with 80% of the samples being positive. The mean counts had no clear pattern related to age. There was an increase of EPG with the increase of temperatures during spring and summer with the peak at midfall. Worm eggs from laying hens were identified as Ascaridia galli. The seasonal differences suggest that higher temperatures might result in an increase of egg survival and sporulation in the environment.

Artículo regular­Muestreo de coccidias y nematodos en aves en pastoreo en el estado de Georgia El objetivo de este estudio fue evaluar y cuantificar el desafío parasitológico en la producción avícola en pastoreo en el estado de Georgia. En el transcurso de un año, se recolectaron muestras fecales de seis parvadas de pavos, 10 parvadas de pollos de engorde y 13 parvadas de gallinas de postura en una granja de pastoreo en intervalos de dos semanas para determinar los conteos de ooquistes de Eimeria y huevos de nematodos. Los recuentos promedio de coccidias fueron 10,198 ooquistes por gramo de heces (OPG) en parvadas de pollos de engorde, 1470 ooquistes por gramo de heces en parvadas ponedoras y 695 ooquistes por gramo de heces en parvadas de pavos. Los promedios en las parvadas de pollos de engorde y pavos fueron más altos en su primera semana en pastoreo. Los conteos en pollos de engorde y ponedoras fueron significativamente más altos en primavera que en invierno y verano. Los recuentos de coccidios en pollos de engorde fueron más bajos que los números publicados en aves criadas de manera convencional, lo que indica que el sistema de rotación de pastizales podría reducir efectivamente la presión de infección. La secuenciación de próxima generación de los productos de PCR mostró la presencia de la mayoría de las especies de Eimeria spp descritas en pollos de engorde, gallinas de postura y pavos. Además, se encontraron unidades taxonómicas operativas (OTU) x, y, z. La frecuencia de especies fue similar para pollos de engorde y gallinas de postura, con la excepción de que Eimeria praecox y las unidades taxonómicas operativas z fueron más comunes en gallinas de postura. En las parvadas de gallinas de postura, el recuento promedio de huevos de helmintos intestinales por gramo de heces (EPG) fue de 509 EPG, con el 80% de las muestras positivas. Los recuentos medios no tenían un patrón claro relacionado con la edad. Hubo un aumento de huevos de helmintos intestinales por gramo de heces con el aumento de las temperaturas durante la primavera y el verano con el pico a la mitad del otoño. Los huevos de helmintos de las gallinas de postura se identificaron como Ascaridia galli. Las diferencias estacionales sugieren que las temperaturas más altas podrían resultar en un aumento de la supervivencia de los huevos y su esporulación en el medio ambiente.

A NEW SPECIES OF EIMERIA (APICOMPLEXA: EIMERIIDAE) FROM THE OLIVE-BACKED POCKET MOUSE, PEROGNATHUS FASCIATUS (RODENTIA: HETEROMYIDAE: PEROGNATHINAE), FROM WYOMING.

Forty-nine olive-backed pocket mice, Perognathus fasciatus were collected during 2011 and 2012 from 4 sites in Wyoming and examined for coccidian parasites. Fifteen (31%) were found to be passing oocysts of a new species of Eimeria Schneider, 1875. Sporulated oocysts of Eimeria fasciata n. sp. are ellipsoidal to ovoidal, 23.3 × 20.7 (19-27 × 17-25) µm, with a shape index of 1.1; they typically contain a single, smooth, bubble-like oocyst residuum. Oocysts possess 1-2 polar granules, lack a micropyle, and are bilayered with a thickness of 1.3 µm. Sporocysts are ovoidal, 10.0 × 8.2 (8-12 × 7-10) µm, with a shape index of 1.2; they contain a sporocyst residuum that appears similar to a cluster of 1-8 grapes. The Stieda body is small, appearing flattened to knobby, and there are no subStieda or paraStieda bodies. This new eimerian represents the only coccidian, to date, reported from P. fasciatus, as well as the only species from any heteromyid rodent in Wyoming.

Global transcriptome landscape of the rabbit protozoan parasite Eimeria stiedae.

BACKGROUND: Coccidiosis caused by Eimeria stiedae is a widespread and economically significant disease of rabbits. The lack of studies on the life-cycle development and host interactions of E. stiedae at the molecular level has hampered our understanding of its pathogenesis. METHODS: In this study, we present a comprehensive transcriptome landscape of E. stiedae to illustrate its dynamic development from unsporulated oocysts to sporulated oocysts, merozoites, and gametocytes, and to identify genes related to parasite-host interactions during parasitism using combined PacBio single-molecule real-time and Illumina RNA sequencing followed by bioinformatics analysis and qRT-PCR validation. RESULTS: In total, 12,582 non-redundant full-length transcripts were generated with an average length of 1808 bp from the life-cycle stages of E. stiedae. Pairwise comparisons between stages revealed 8775 differentially expressed genes (DEGs) showing highly significant description changes, which compiled a snapshot of the mechanisms underlining asexual and sexual biology of E. stiedae including oocyst sporulation between unsporulated and sporulated oocysts; merozoite replication between sporulated oocysts and merozoites; and gametophyte development and gamete generation between merozoites and gametocytes. Further, 248 DEGs were grouped into nine series clusters and five groups by expression patterns, and showed that parasite-host interaction-related genes predominated in merozoites and gametocytes and were mostly involved in steroid biosynthesis and lipid metabolism and carboxylic acid. Additionally, co-expression analyses identified genes associated with development and host invasion in unsporulated and sporulated oocysts and immune interactions during gametocyte parasitism. CONCLUSIONS: This is the first study, to our knowledge, to use the global transcriptome profiles to decipher molecular changes across the E. stiedae life cycle, and these results not only provide important information for the molecular characterization of E. stiedae, but also offer valuable resources to study other apicomplexan parasites with veterinary and public significance.

First report of Eimeria and Entamoeba infection in alpacas (Vicugna pacos) in Shanxi Province, northern China.

Intestinal protozoa Eimeria and Entamoeba can infect many animal species including alpacas. However, data on the prevalence and pathogenicity of species of the two genera Eimeria and Entamoeba in alpacas in China is scarce. The current study was carried out to investigate the prevalence of Eimeria and Entamoeba in alpacas in two cities (Taiyuan and Xinzhou) in Shanxi Province, northern China, using PCR-based approaches. Eimeria spp. were only found in Taiyuan city, and the overall prevalence was 1.64%. All samples collected from male alpacas were PCR-negative for Eimeria. Four Eimeria-positive samples were tested positive as Eimeria lamae. The molecular prevalence of Entamoeba in alpacas was 18.03% (66/366), including 16.39% (50/305) in alpacas from Taiyuan city and 26.23% (16/61) from Xinzhou city, respectively. The Entamoeba prevalence in male alpacas (25.00%) was significantly higher than that in female alpacas (15.69%). Entamoeba bovis was the predominant species, and no Entamoeba histolytica infection was detected. Nine unique SSU rRNA gene sequences of Entamoeba were obtained which formed a new cluster. The results showed that sex and location might be the risk factors associated with prevalence of Eimeria spp., and sex might be the risk factor associated with prevalence of Entamoeba spp.. This is the first report of Entamoeba in alpacas worldwide. These findings expand our understanding of the prevalence and genetic diversity of Eimeria and Entamoeba in alpacas.

Molecular detection of Eimeria species and Clostridium perfringens in poultry dust and pooled excreta of commercial broiler chicken flocks differing in productive performance.

Necrotic enteritis and coccidiosis are the most economically detrimental enteric diseases of broiler chickens. This study aimed to investigate the association of DNA load of Clostridium perfringens, netB, and five Eimeria species (E. brunetti, E. maxima, E. necatrix, E. acervulina and E. tenella) in poultry house dust and pooled excreta with flock productive performance. The dust and pooled excreta from the floor were collected weekly at days 7, 14, 21, 28 and 35 of chicken age from 16 flocks of eight farms from two Australian integrator companies. The farms were ranked as high or low performers by each integrator according to the production performance of studied flocks. Eimeria tenella and necatrix were not detected in any farm while E. brunetti was detected in a low-performance farm and netB was detected in a high-performance farm. C. perfringens, E. acervulina and E. maxima DNA were detected on all farms with no significant differences in DNA load between high and low-performance farms or companies. The lack of association of pathogen DNA load and farm performance is possibly due to overall low to moderate pathogen DNA load detected in this study. Further studies on a larger number of farms are needed to determine whether these population level measurements of key pathogens based on PCR detection of nucleic acids are correlated with performance variables.

Prevalence of Eimeria parasites in the Hubei and Henan provinces of China.

Coccidiosis is an intestinal parasitic disease that causes huge economic losses in the poultry industry globally. Henan and Hubei, as important poultry production provinces in China, have great pressure for the prevention and control of chicken coccidiosis. In order to obtain information on the local prevalence of Eimeria species, we used an internal transcribed spacer 1 (ITS1) sequence of ribosomal DNA to identify the species from 318 fresh fecal samples. The fecal samples and the data relating to farm information were collected from 137 farms in Hubei and Henan provinces. As shown by genus-specific PCR results, the positivity rate of Eimeria was 97.17% (309/318), and the most common species were Eimeria mitis (66.67%), E. tenella (46.86%), and E. necatrix (41.51%). Then, we analyzed the correlation between the background information of each sample and the PCR identification results, which showed that indigenous farms in Henan province were at the greatest risk of harboring highly pathogenic Eimeria species and a larger proportion of such farms were positive for E. necatrix, the most pathogenic species. The results of this study showed that chicken coccidia was widespread, which provides important insights into the control of chicken coccidiosis in this region.

Analysis of potential risk factors of caprine coccidiosis.

Caprine coccidiosis represents a major concern in goat industry worldwide causing high economic losses, mainly due to poor weight gains, treatment costs and even death of kids. Therefore, analyses on risk factors involved in the establishment and outcome of this disease are important to improve infection status and herd management. In Portugal, approximately 400,000 goats are reared every year. The majority of them are allocated in the southern part of the country. In the current study, a representative number of animals (382 goats) and herds (37 herds) from Alentejo were monitored. Faecal samples from five young and five adult goats of each farm were collected rectally and examined individually by using the modified McMaster technique for oocyst counts (oocyst per gram of faeces - OPG). Additionally, a questionnaire on the husbandry conditions and anti-parasitic treatments was performed. Overall, a prevalence of Eimeria infections of 99% and 100% was detected on animal and herd level, respectively. Eimeria arloingi and E. ninakohlyakimovae as the two most pathogenic caprine species also constituted the most prevalent ones (34.91% and 29.90%, respectively), both in the young and the adult goat groups. As expected, OPG vales were significantly higher (p = 0.031) in young than in adult goats. Furthermore, risk factor analyses revealed young age, higher total number of animals on the farm and outdoor kidding as strongly associated with a higher risk of caprine coccidiosis. Overall, the high prevalence of pathogenic Eimeria species on Portuguese goat farms in combination with the occurrence of related risk factors strengthen the importance of coccidiosis control in caprine industry.

Correlation Between Clostridium perfringens Alpha- and NetB-Toxin and Chick Mortality in Commercial Broiler Farms During Different Anticoccidial Control Programs.

The purpose of the present study was to determine whether a correlation existed between chick mortality and the presence of Clostridium perfringens alpha-toxin and NetB-toxin genes (cpa and netB) in C. perfringens recovered from litter in commercial broiler houses. Because coccidiosis predisposes chickens to necrotic enteritis, the concentration of Eimeria oocysts in these samples was measured, and the numbers were used in similar correlation analyses. Litter samples were collected at 0, 2, and 4 wk growout from six broiler farms (18 houses total) during an anticoccidial drug (ACD) control program and from nine broiler farms (23 houses total) during an Eimeria vaccine (VAC) control program. Of these, litter samples were collected from five farms during both ACD and VAC programs. The litter samples were processed for Eimeria oocyst and C. perfringens spore enumerations by standard parasitologic and microbiologic techniques. DNA was also extracted for C. perfringens DNA for PCR detection of genes coding for alpha- and NetB-toxin. A general trend during the ACD programs was a transient decrease in both Eimeria maxima and non-E. maxima (Eamipt) numbers at 2 wk growout. The pattern was slightly different during VAC with E. maxima and Eamipt levels increasing over time. Average concentrations of C. perfringens in litter were highest at 2 wk (∼105-106 spores/g) during ACD and at placement during VAC (∼105-106 spores/g). During the ACD program, a strong correlation was observed between 0 and 3-wk chick mortality and the presence at placement (0 wk) of netB (r = 0.42-0.48) or cpa (r = 0.55-0.67). A very strong correlation was observed in 0-5-wk chick mortality and the presence of netB at 4 wk growout (0.73-0.95). During a VAC program, a strong correlation was only observed between the presence of netB at placement and 0-1-wk chick mortality (r = 0.67).

Study on the prevalence and genetic diversity of Eimeria species from broilers and free-range chickens in KwaZulu-Natal province, South Africa.

This study was conducted from January to October 2018 with the objective to determine the prevalence and genetic diversity of Eimeria species in broiler and free-range chickens in KwaZulu-Natal province, South Africa. A total of 342 faecal samples were collected from 12 randomly selected healthy broiler chicken farms and 40 free-range chickens from 10 different locations. Faecal samples were screened for the presence of Eimeria oocysts using a standard flotation method. The species of Eimeria isolates were confirmed by amplification of the internal transcribed spacer 1 (ITS-1) partial region and sequences analysis. Among broiler and free-ranging chickens, 19 out of 41 pens (46.3%) and 25 out of 42 faecal samples (59.5%) were positive for Eimeria infection. Molecular detection revealed the following species: Eimeria maxima, Eimeria tenella, Eimeria acervulina, Eimeria brunetti and Eimeria mitis in all the samples screened. Similarly, polymerase chain reaction assays specific for three cryptic Eimeria operational taxonomic units were negative for all the samples. Phylogenetic analysis of the ITS-1 sequences supported species identity with the greatest variation detected for E. mitis. This study provides information on the range and identity of Eimeria species, and their genetic relatedness, circulating in commercially reared broilers and free-ranging chickens from different locations in KwaZulu-Natal province.

First record of clinical coccidiosis (Eimeria ovinoidalis) in adult sheep from northwestern Argentina.

Coccidiosis of sheep is an intestinal infection caused by protozoa of the genus Eimeria. An outbreak of the disease in adult sheep from Salta province, northwestern Argentina, was studied to establish its clinical, epidemiological, pathological and etiological aspects. The affected animals were part of a flock of 20 sheep brought from Formosa province about 10 days before. Most sheep (80% incidence) showed hemorrhagic diarrhea, dehydration and loss of body condition; six of them died and two that became permanently recumbent were euthanized. Three necropsied sheep showed mild mesenteric lymphadenomegaly, diffuse proliferative enteritis in the small and large intestines, and mucosal thickening. Histopathological studies exhibited diffuse proliferative enteritis and presence of structures compatible with intracellular coccidia at different stages of development. Parasitological studies (n = 12) resulted in an average of 16,636.6 (± 15,266.8) Eimeria oocysts per gram of feces (range 1680-46,400). Taxonomy of Eimeria species based on analysis of sporulated oocysts derived from 4 fecal samples (n = 100 oocyst per sample) showed, on average, a high prevalence of E. ovinoidalis (61.5%), followed by E. parva (27.2%), and lower proportions of E. crandallis (5.3%), E. ahsata (3.2%) and E. intricata (2.8%). Clinical and pathological findings confirmed the diagnosis of coccidiosis in the affected sheep; parasitological results showed that E. ovinoidalis was the main species responsible for the clinical signs. Clinical coccidiosis is considered unusual in adult sheep, but the present case shows that under favorable environmental and/or management conditions, this infection may be highly deleterious for adult sheep.

Modulation of the intestinal microbiota of broilers supplemented with monensin or functional oils in response to challenge by Eimeria spp.

The objective of this study was to evaluate the effect of supplementation with 100ppm sodium monensin or 0.15% of a blend of functional oils (cashew nut oil + castor oil) on the intestinal microbiota of broilers challenged with three different Eimeria spp. The challenge was accomplished by inoculating broiler chicks with sporulated oocysts of Eimeria tenella, Eimeria acervulina, and Eimeria maxima via oral gavage. A total of 864, day-old male broiler chicks (Cobb) were randomly assigned to six treatments (eight pens/treatment; 18 broilers/pen) in a 3 × 2 factorial arrangement, composed of three additives (control, monensin or blend), with or without Eimeria challenge. Intestinal contents was collected at 28 days of age for microbiota analysis by sequencing 16s rRNA in V3 and V4 regions using the Illumina MiSeq platform. Taxonomy was assigned through the SILVA database version 132, using the QIIME 2 software version 2019.1. No treatment effects (p > 0.05) were observed in the microbial richness at the family level estimated by Chao1 and the biodiversity assessed by Simpson's index, except for Shannon's index (p < 0.05). The intestinal microbiota was dominated by members of the order Clostridiales and Lactobacillales, followed by the families Ruminococcaceae, Bacteroidaceae, and Lactobacillaceae, regardless of treatment. When the controls were compared, in the challenged control group there was an increase in Erysipelotrichaceae, Lactobacillaceae, Bacteroidaceae, Streptococcaceae, and Peptostreptococcaceae, and a decrease in Ruminococcaceae. Similar results were found for a challenged group that received monensin, while the blend partially mitigated this variation. Therefore, the blend alleviated the impact of coccidiosis challenge on the microbiome of broilers compared to monensin.

Protozoan and Helminth Infections of Aviary-Reared Alectoris rufa (Galliformes: Phasianidae) before Releasing for Hunting in the Czech Republic: Infection Dynamics and Potential Risks.

Age-related dynamics of protozoan and helminth infections in the red-legged partridge, Alectoris rufa, were studied in an intensive breeding aviary in the Czech Republic before releasing birds for hunting purposes. Pooled fecal samples (n = 900) were examined over 3 rearing seasons (2012-2014). A total of 4 protozoan species, Cryptosporidium baileyi, Cryptosporidium meleagridis, Eimeria kofoidi, and Eimeria legionensis, and 4 helminths, Capillaria phasianina, Eucoleus perforans, Heterakis gallinarum, and Syngamus trachea, was found. The most common parasite was S. trachea (prevalence 20-26%) in dead birds (n = 99), which represents a high risk for breeders. Co-occurrence of protozoans and helminths indicated similarities in infection dynamics throughout the 3 breeding seasons. Mixed infections of Cryptosporidium baileyi and C. meleagridis with other parasitic species are reported for the first time. Our findings provide new insights into breeding of A. rufa and may help to improve the efficacy of disease control strategies and prevention, especially with the potential for spreading of parasitic infections to wildlife through released birds into open areas.

Coccidian Parasites of Flying Squirrels, Glaucomys spp. (Rodentia: Sciuridae), from Alaska and Arkansas, with a Description of a New Species of Eimeria (Apicomplexa: Eimeriidae).

New World flying squirrels, Glaucomys spp., are nocturnal arboreal sciurid rodents that have been previously surveyed for coccidial parasites. To date, 4 species of Eimeria have been reported from 2 species of Glaucomys. Here we report 2 species of eimerians from southern flying squirrels (Glaucomys volans) and the endemic Prince of Wales flying squirrel (Glaucomys sabrinus griseifrons). Oocysts of Eimeria dorneyi Levine and Ivens were found to be passing in the feces of 4 G. s. griseifrons from Alaska and a new species of Eimeria was present in feces from 6 G. volans from Arkansas. Oocysts of Eimeria hnidai n. sp. are ellipsoidal with a bilayered wall, measure 23.7 × 13.7 µm, and have a length/width (L/W) ratio of 1.7; a micropyle and oocyst residuum are absent but polar granule(s) are present. Sporocysts are ellipsoidal-elongate and measure 11.8 × 4.9 µm, L/W 2.2; Stieda body is present but sub-Stieda and para-Stieda bodies are absent. The sporocyst residuum is composed of small indistinct granules along the edge or in the center of the sporocyst. This is the first coccidian reported from G. volans from Arkansas as well as the initial coccidian (E. dorneyi) reported from G. s. griseifrons from Alaska. We also provide a summation of the coccidia known from North American flying squirrels.

Presence of eimerid oocysts in faeces of a quarantined dog in Iceland is explained by coprophagic behaviour prior to its importation. Case report.

BACKGROUND: All dogs imported into Iceland must undergo mandatory quarantine in a special station before introduction into the country. A faecal sample is collected from the first stool passed by the dog in this station and subsequently examined for the presence of intestinal parasite stages. CASE PRESENTATION: In May 2019 unsporulated oocysts were detected in faeces from a 7-year-old household dog that had been imported from Sweden. Most of the oocysts studied strongly resembled those of Eimeria canis Wenyon, 1923. As this species is not valid, the purpose of the present article was to identify the correct species and examine their possible origin. Studies confirmed the presence of two distinct unsporulated oocyst morphotypes in the faeces; measurements and photomicrographs confirmed their identification as Eimeria magna Pérard, 1925 and Eimeria stiedai (Lindemann, 1865) Kisskalt and Hartmann, 1907, both common parasites of European rabbits, Oryctolagus cuniculus (L., 1758). When the owner of the dog was questioned about the food administrated to the dog prior to its import to Iceland, it turned out that it had exclusively been fed dry dog food pellets. However, the owner also reported that on the morning prior to transportation to Iceland, the dog was allowed to move freely in a grassland area where rabbits are common and heaps of their faeces are present. Furthermore, the owner confirmed that the dog consumed rabbit faeces that morning. CONCLUSION: It is believed that this coprophagic behaviour can explain the detection of rabbit eimerids in the dog's faeces, and that such behaviour must be taken into consideration by veterinarians and other diagnostic personnel when they detect atypical cysts or eggs during coprological examinations.