Molecular detection of Toxoplasma gondii and Neospora caninum in seabirds collected along the coast of Santa Catarina, Brazil.

Toxoplasma gondii and Neospora caninum are two closely related protozoans that infect a wide range of animals, including birds. However, the occurrence of N. caninum and T. gondii in seabirds is unknown. Therefore, this study aimed to determine the presence of T. gondii and N. caninum DNA in tissue samples of seabirds. Tissue samples of the pectoral muscles, heart, and brain were collected from 47 birds along the coastline of Santa Catarina State, SC, Brazil. The DNA was extracted from the tissues and screened using nested-PCR (nPCR) targeting internal transcribed spacer 1 (ITS1). T. gondii DNA was detected in tissues from seven seabirds (7/47, 14.8%), kelp gull (Larus dominicanus) (5/21), and Manx shearwater (Puffinus puffinus) (2/8). N. caninum DNA was detected in tissues of nine seabirds (9/47, 19.1%), the kelp gull (L. dominicanus) (4/21), Manx shearwater (P. puffinus) (2/8), neotropic cormorant (Phalacrocorax brasilianus) (1/4), brown booby (Sula leucogaster) (1/5), and white-chinned petrel (Procellaria aequinoctialis) (1/1); however, no co-infection was observed. In conclusion, this study showed the circulation of N. caninum and T. gondii in seabirds along the coastline of Santa Catarina State. Further studies are required to clarify the role of these birds in the epidemiology of neosporosis and toxoplasmosis.

The seroprevalence and risk factors for exposure to Neospora caninum and Neospora hughesi in Ontario broodmares.

The seroprevalence and risk factors for exposure to Neospora caninum and Neospora hughesi in broodmares in Ontario were investigated. Sixty of the 219 (27.4%) study broodmares were seropositive for N. caninum and 65/219 (29.7%) for N. hughesi with cut-offs of ≥1:40 and ≥1:160, respectively. Thirty-one of 63 participating farms (49.2%) had at least 1 broodmare seropositive for N. caninum. Thirty-three of the 63 (52.4%) participating farms had at least 1 broodmare positive for N. hughesi. Risk factors for N. caninum included presence of farm dogs (OR = 6.70; 95% CI = 2.14-20.97; p = 0.001), and high stocking density (OR = 2.83; 95% CI = 1.27-6.30; p = 0.011). Presence of livestock, excluding cattle, was associated with reduced risk of exposure (OR = 0.17; 95% CI = 0.06-0.53; p = 0.002). The only risk factor for exposure to N. hughesi was feeding hay on the ground in the paddock (OR = 4.31; 95% CI = 1.65-11.22; p = 0.003). This study demonstrated widespread exposure to Neospora spp. in broodmares in Ontario.

CHARACTERIZATION OF EIMERIA LANCASTERENSIS AND EIMERIA ONTARIOENSIS (APICOMPLEXA: EIMERIIDAE) FOUND IN EASTERN GRAY SQUIRRELS, SCIURUS CAROLINENSIS (RODENTIA: SCIURIDAE), FROM ARKANSAS AND OKLAHOMA.

We report the morphological characteristics of oocysts of Eimeria lancasterensisJoseph, 1969, collected from 6 of 6 (100%) eastern gray squirrels, Sciurus carolinensis, collected in Arkansas (n = 3) and Oklahoma (n = 3), and Eimeria ontarioensisLee and Dorney, 1971, recovered from an individual of S. carolinensis from Arkansas. Oocysts of E. lancasterensis were ovoidal to ellipsoidal, measuring (L × W) 24.0 × 14.6 (18-29 × 12-16) µm; shape index (L/W) was 1.6 (1.3-1.8). A micropyle and an oocyst residuum were absent, but up to 2 polar granules were present. Oocysts of E. ontarioensis were piriform and measured 40.6 × 26.0 (37-44 × 23-28); L/W was 1.6 (1.5-1.7). These oocysts possessed a distinct micropyle and rarely a polar granule but lacked an oocyst residuum. The DNA was isolated from both eimerians, and the 18S rDNA genetic markers were PCR-amplified, cloned, sequenced, and analyzed. To our knowledge, this study represents the first time 18S DNA sequence data have been generated from E. lancasterensis and E. ontarioensis found in North American sciurid hosts, as well as new geographic distribution records for these coccidians. In addition, we also include a tabular summary of these 2 species of Eimeria from Sciurus spp. worldwide, with information on their hosts, distribution, and taxonomically important morphological characteristics, including key measurements of oocysts and sporocysts.

New insights on the phylogeography of Hepatozoon canis in Brazil.

This study aimed to molecularly characterize the Hepatozoon spp. infecting domestic and wild dogs in Brazil. A total of 22 whole blood samples tested positive for Hepatozoon spp., and five samples were sequenced for the 18S rDNA gene from H. canis after PCR amplification with four primer sets. Phylogenetic analysis using Bayesian inference showed that the three H. canis isolates from domestic dogs were not monophyletic; however, they were more closely related to each other than to other H. canis sequences. The isolate from the hoary fox (Lycalopex vetulus) was phylogenetically more distant. Two haplotype networks were constructed, identifying 10 haplotypes of H. canis in Brazil, with H10 constituting the largest group. It contains nine isolates, including three from domestic dogs. The H5 haplotype grouped the sequence of L. vetulus with two additional sequences from hosts Tapirus terrestris and L. vetulus, representing the sole haplotype with wild hosts. Bayesian analysis suggested the possible existence of two genetic groups of H. canis in Brazil, indicating gene flow of this agent within the country. These findings contribute valuable insights for a more comprehensive understanding of the molecular diversity of Hepatozoon spp. in Brazil and may help in the development of effective control measures.

Prediction of coccidiosis prevalence in extensive backyard chickens in countries and regions of the Horn of Africa.

Coccidiosis is one of the leading morbidity causes in chickens, causing a reduction of body weight and egg production. Backyard chickens are at risk of developing clinical and subclinical coccidiosis due to outdoor housing and scavenging behaviour, jeopardizing food security in households. The objectives of this study were to estimate clinical prevalence of coccidiosis at country and regional levels in the Horn of Africa in extensive backyard chickens. A binomial random effects model was developed to impute prevalence of coccidiosis. Previously gathered prevalence data (n = 40) in backyard chickens was used to define the model. Precipitation (OR: 1.09 (95% CI: 1.05-1.13) and the presence of seasonal rainfall (OR: 1.85, 95% CI: 1.27-2.70) significantly increase prevalence. Results showed an overall prevalence of coccidiosis in the Horn of Africa of 0.21 (95% CI: 0.15-0.29). Ethiopia, the Republic of South Sudan and Kenya showed the highest prevalence and Djibouti the lowest. Significant differences between Djibouti and the countries with highest prevalence were found. However, no evidence of a significant difference between the rest of the countries. Kenya and Ethiopia showed larger prevalence differences between regions. Results could assist with the targeting of testing for coccidiosis, the observation for clinical disease of chickens living in specific regions and as a baseline for the evaluation of future control measures.

Seroprevalence of Neospora caninum in pet cats, dogs and rabbits from urban areas of Poland.

BACKGROUND: Neospora caninum (N. caninum) has a broad intermediate host range and might cause multisystemic lesions in various species of animals. Dogs are both intermediate and definitive hosts of the parasite and play a crucial role in the horizontal transmission of this protozoan to other animals. Cats and rabbits could be sensitive to infection with N. caninum, however, clinical symptoms and the exact route of infection in these species are unknown. The epidemiology of N. caninum in cats and rabbits has been barely researched, and there is no published record of the seroprevalence of N. caninum infection in these species in Poland. Thus, the present study aimed to determine the frequency of seroreagents for N. caninum within pet dogs, cats and rabbits from urban areas of Poland and to identify possible risk factors for these animals. RESULTS: In total, serum samples from 184 cats (Felis catus), 203 dogs (Canis familiaris) and 70 rabbits (Oryctolagus cuniculus) were used in the study. The seroprevalence of anti-N. caninum antibodies in dogs and cats reached 1.0% (2/203; 95% CI: 0.3-3.5) and 3.3% (6/184; 95% CI: 1.5-6.9), respectively. No significant differences in seroprevalence regarding age group, gender, symptoms or sampling location were found. All 70 samples from pet rabbits were negative for anti-N. caninum antibodies. CONCLUSIONS: The seroprevalence rates of N. caninum in dogs and cats in the present study were low, however, our results confirmed N. caninum circulates among dog and cat populations in Poland, and neosporosis should be included in the differential diagnosis of neuro-muscular disorders in these species. This is the first serological survey of N. caninum in European pet cats and rabbits. The role of pet rabbits in N. caninum epidemiology and circulation in Poland is marginal.

Coinfection of the gut with protozoal and metazoal parasites in broiler and laying chickens.

The chicken business faces substantial economic losses due to the risk of parasitic coinfection. Because the current study aimed to investigate enteric parasitic coinfections problems among the suspected examined chicken farms, samples were collected during the field investigation from suspected freshly dead birds, clinically diseased, apparently healthy, and litter samples for further laboratory parasitological, histopathological, and immunological examinations. Variable mortalities with various clinical indicators, such as ruffled feathers, weight loss, diarrhea of various colors, and a decline in egg production, occurred on the farms under investigation. In addition, the treatment protocols of each of the farms that were evaluated were documented and the m-RNA levels of some cytokines and apoptotic genes among the infected poultry have been assessed. The prevalence rate of parasitic coinfection in the current study was found to be 8/120 (6.66%). Parasitological analysis of the samples revealed that they belonged to distinct species of Eimeria, cestodes, and Ascaridia galli. When deposited, A. galli eggs were nonembryonated and ellipsoidal, but cestodes eggs possessed a thin, translucent membrane that was subspherical. Eimeria spp. oocysts in layer chickens were identified as Eimeria acervulina and Eimeria maxima in broiler chickens. Our findings proved that coinfection significantly upregulated the IL-1ß, BAX, and Cas-3 genes. Conversely, the IL-10, BCL-2, and AKT mRNA levels were downregulated, indicating that nematode triggered apoptosis. The existence of parasite coinfection was verified by histological investigation of the various intestinal segments obtained from affected flocks. A. galli and cestodes obstructed the intestinal lumen, causing different histological alternations in the intestinal mucosa. Additionally, the lamina propria revealed different developmental stages of Eimeria spp. It was determined that parasite coinfection poses a significant risk to the poultry industry. It was recommended that stringent sanitary measures management methods, together with appropriate treatment and preventative procedures, be employed in order to resolve such issues.

Identification of Eimeria spp. in domestic chickens raised in alternative poultry production systems in the State of São Paulo, Brazil.

The objective of this study was to identify Eimeria spp. in alternative poultry production systems (APPS) in the State of São Paulo, Brazil. Fecal samples (168) and DNA extracted from fecal samples obtained in APPS located in different Municipalities in the State of São Paulo (93) were examined by microscopy or genera-specific PCR (ITS-1 locus). Samples positive for Eimeria spp. were examined using Eimeria lata, Eimeria nagambie, and Eimeria zaria species-specific PCR protocols (ITS-2 locus) and another E. lata-specific PCR (candidate IMP1 genomic locus) followed by molecular cloning (E. lata and E. zaria ITS-2 amplicons) and genetic sequencing. All positive DNA samples were also submitted to genera-specific nested PCR (18S rRNA gene) followed by next-generation sequencing to identify Eimeria spp. Eimeria nagambie, E. zaria, and Eimeria sp. were identified by ITS2-targeted species-specific PCRs and genetic sequencing. Next-generation sequencing identified, in order of prevalence: E. nagambie; Eimeria acervulina; Eimeria mivati; Eimeria praecox; Eimeria brunetti; Eimeria mitis; Eimeria sp.; Eimeria maxima; E. zaria, and Eimeria necatrix/tenella. Our results confirmed, for the first time in Brazil, the identification of E. nagambie, E. zaria, and Eimeria spp. ITS-2 and 18S rRNA gene sequences not yet described in Brazil.

EPIDEMIOLOGY OF TOXOPLASMA GONDII AND NEOSPORA CANINUM INFECTION IN GOATS FROM AGUASCALIENTES, MEXICO.

The objective of this study was to describe the epidemiology of Toxoplasma gondii and Neospora caninum infection by estimating seroprevalence and its association with certain risk factors in goats from Aguascalientes, Mexico. A total of 150 blood samples was taken from 10 farms and serum samples were subjected to enzyme-linked immunosorbent assay indirect test to detect T. gondii and N. caninum antibodies; the association between seroprevalence and some potential risk factors was estimated through logistic regression analysis. The general seroprevalence for T. gondii was 12.6%, observed in the farms in a range of 6.6 to 60%, finding seropositive animals in 80% of them; for N. caninum the seroprevalence was 3.3% and in farms a range of 6.6 to 13.3% was identified and 30% of them had at least 1 seropositive animal. The coinfection was 0.66%. The risk analysis for T. gondii identified a history of abortions (odds ratio 9.25) as a factor associated with seroprevalence; for N. caninum, no risk factor was identified.

A NEW EIMERIA SPECIES (APICOMPLEXA: EIMERIIDAE) PARASITIZING COMMERCIAL CHUKAR PARTRIDGES (ALECTORIS CHUKAR) IN SOUTHERN ONTARIO.

A commercial producer hatching and rearing chukar partridges (Alectoris chukar) in Ontario, Canada had flocks experiencing coccidiosis. Microscopic analysis of Eimeria species isolated from a field sample indicated the presence of 2 distinct oocyst morphotypes; the most abundant species was determined to be Eimeria chapmani, based on oocyst morphology and sequence-based genotyping, and the less abundant, second Eimeria sp. was an undescribed parasite. Oocysts of the unknown Eimeria sp. were large and oval-shaped; dimensions averaged 27.9 µm by 17.0 µm (shape index = 1.65 µm). Oocysts contained at least 1 polar granule and 4 almond-shaped sporocysts with average dimensions measuring 12.5 µm by 6.9 µm (shape index = 1.83). Each sporocyst featured a Stieda body, sub-Stieda body, and sporocyst residuum; a sporocyst contained 2 sporozoites that each possessed a small anterior refractile body and a larger posterior refractile body. Virtually all oocysts sporulated after 24 hr when suspended in potassium dichromate at room temperature (22 C) on a rotary platform. Experimental infections with various doses of oocysts demonstrated elevated parasite shedding from birds gavaged with higher challenge doses; fecundity generally decreased in heavier infections. The approximate prepatent period of the parasite was 4-5 days (unsporulated oocysts observed histologically at 90 hr postinfection and in feces by day 5) and patency lasted until day 12 postinfection. To characterize the endogenous development of the Eimeria sp., tissues were collected at 8 regions along the intestinal tract (including the ceca and rectum) every 6 hr throughout the estimated prepatent period. Parasites were observed to infect the descending and ascending duodenum, midjejunum, proximal and distal ileum, and the ceca. The endogenous stages identified included intracellular sporozoites, 3 generations of merogony, and gametogonic stages. Sequences of the mitochondrial genome (GenBank MW934555) and nuclear 18S ribosomal DNA (GenBank MW934259) were obtained using polymerase chain reaction amplification for Sanger sequencing, and these were unique from all published sequences on GenBank. Molecular data, in conjunction with the unique biology of the Eimeria sp. isolated from the chukar partridge flock, support that this coccidium is new to science.

Serosurvey and associated risk factors for Neospora caninum infection in Egyptian water buffaloes (Bubalus bubalis).

Neosporosis is a parasitic disease that causes reproductive disorders in animals, making it a barrier to maximum efficiency. The purpose of this study was to determine the seroprevalence of Neospora caninum (N. caninum) antibodies in water buffaloes from four governorates in northern Egypt. A commercial indirect-ELISA test was used to detect antibodies against N. caninum in the serum of 450 water buffaloes. The total seroprevalence of N. caninum in water buffaloes from Egypt was 31.3%, and the highest prevalence was observed in Gharbia governorate. The identified risk factors for N. caninum infections in water buffaloes were sex (OR = 1.96, 95%CI: 1.22-4.17), buffaloes more than 4 years of age ( OR = 5.80, 95%CI: 2.26-14.86), abortion in second trimester (OR = 16.48, 95%%CI: 2.99-34.03), history of abortion (OR = 3.45, 95%CI: 1.58-7.52) and contact with dogs (OR = 2.55, 95%CI: 1.51-4.32). Thus, more studies are needed to determine the role of buffaloes in the epidemiology of neosporosis in Egypt.

Serological and molecular detection of Toxoplasma gondii and Neospora caninum in free-ranging rats from Nagpur, India.

Toxoplasma gondii and Neospora caninum are cyst-forming coccidian parasites that infect both wild and domestic non-felids as intermediate hosts, with rodents serving as important reservoir hosts during their life cycles. This study was aimed at investigating T. gondii and N. caninum infections and identifying factors favouring T. gondii infection in free-ranging rats from India. A total of 181 rodents were trap-captured, and blood and brain samples were subsequently collected for serological and molecular examination of T. gondii and N. caninum. Antibodies against T. gondii and N. caninum were detected by MAT/NAT and IFAT in 13.8% (25/181) and 1.65% (3/181) of rodents, respectively. All three N. caninum samples positive by NAT/IFAT were also positive for ELISA, while for T. gondii, 19 of 25 MAT/IFAT positive samples were also positive for ELISA. The antibody titers (MAT/NAT/IFAT) of rodents seropositive for T. gondii ranged from 25 to 400, while those of rats seropositive for N. caninum ranged from 25 to 100. Also, using PCR, DNA from T. gondii (B1 gene) and N. caninum (NC5 gene) was found in 2.76% (5/181) of brain samples and 0.55% (1/181) of brain samples. All PCR positive samples were also seropositive. No mixed infections were observed in the serological and molecular detections. A Chi-square analysis revealed that older rats and rats living in urban areas are significantly associated with T. gondii infection; however, rodent species, gender, location, habitat types, and seasonality were statistically nonsignificant. Overall, this study demonstrated that T. gondii was widely distributed while N. caninum was less prevalent among free-ranging rats in the studied area.

First detection and characterisation of Eimeria zaria in European chickens.

The global poultry industry has experienced dramatic growth in recent decades, increasing the significance of pathogens of chickens. Protozoan parasites of the genus Eimeria can cause the disease coccidiosis, compromising animal health and welfare, and incurring significant annual costs. Seven Eimeria species have long been recognised to infect chickens, supplemented by three new candidate species first reported from Australia in 2007/8. Named Eimeria lata, Eimeria nagambie and Eimeria zaria, one or more of these new species have been reported in Australia, several countries in sub-Saharan Africa, India, Venezuela, and most recently the United States of America, but none have been detected in Europe. Here, a panel of 56 unvaccinated broiler chicken farms were sampled in the final week of production from France, Greece, Italy, the Netherlands, the Republic of Ireland, and the United Kingdom to assess the occurrence of all ten Eimeria species using specific polymerase chain reaction (PCR). Overall, 39 of 56 (69.6%) farms were found to host at least one species. Eimeria acervulina, E. tenella, and E. maxima were most common, with E. mitis and E. praecox also widespread. Eimeria necatrix was detected on one farm in France, while E. brunetti was not detected. Eimeria zaria was detected for the first time in Europe, appearing in Greece and Italy (one occurrence each). New primers were designed to confirm detection of E. zaria and provide template for phylogenetic comparison with the reference isolate from Australia. Detection of E. zaria in Europe reinforces the importance of integrated control for coccidiosis given the lack of protection induced by current anticoccidial vaccines.

Seroprevalence and associated factors with Neospora caninum infection in sheep and goats slaughtered in the state of Paraíba, Brazil.

The objective was to describe the seroprevalence of anti-Neospora caninum antibodies in goats and sheep slaughtered in the state of Paraíba and to identify possible associated factors with the infections. Two hundred twenty-nine samples from goats and two hundred five from sheep were analyzed by Indirect Immunofluorescence Reaction (IFAT) using a cutoff point of 1:50. The presence of anti-N. caninum antibodies was identified in 28.4% (65/ 229; 95% Confidence Interval: 22.6-34.2) of the goat samples and in 12.7% (26/ 205; 95% CI: 8.2 - 17.2) of the sheep samples. Contact between goats and dogs (Odds ratio 4.81; CI 1.13 - 2.67; p = 0.041) and cattle (OR. 1.87; CI 1.13 - 2.67; p = 0.002) was identified as a risk factor for goats and contact between sheep and dogs (OR 2.32; CI 1.58 - 3.14; p = 0.026) and history of abortion (OR 1.94; CI 1.28 - 2.90; p = 0.001) was considered a risk factor for sheep. The study revealed a high seroprevalence of anti-N. caninum antibodies in slaughtered goats and sheep in Paraíba. Risk factors such as contact with dogs/cattle and abortion history underscore the need for preventive measures to control infection and enhance animal health management.

Evaluation of frequency of antibodies against Neospora caninum, Toxoplasma gondii and Brucella melitensis, risk factors and spatial distribution of infection in goat and sheep flocks from Argentina.

Neospora caninum, Toxoplasma gondii and Brucella melitensis are pathogens that cause abortion in small ruminants. Besides, B. melitensis and T. gondii are zoonotic pathogens. The aim of this study was to describe the frequency of antibodies against N. caninum, T. gondii and B. melitensis in sheep and goats from three provinces of the center region of Argentina. In addition, the spatial distribution of the infected flocks/herds and risk factors were evaluated. A cross-sectional study was conducted from 2015 through 2016. Serum samples from 4783 goats and 1524 sheep from 186 goat, 51 sheep and 38 mixed flocks/herds were analyzed. Competitive inhibition enzyme-linked immunosorbent assay (ciELISA) and indirect fluorescent antibody test (IFAT) were performed for detection of antibodies against N. caninum and IFAT for T. gondii. The buffered plate antigen test and complement fixation test were performed for detection of antibodies against B. melitensis. The frequency of anti-T. gondii antibodies was 41.2% and 29.7% for sheep and goats, respectively. The frequency of anti-N. caninum antibodies was 17.2% and 14% for sheep and goats, respectively. About 97.1% of the sheep flocks, 79.4% of the goat herds and the 91.3% of the mixed flocks had seropositive animals to T. gondii. About 61.8% of the sheep flocks, 58% of the goat herds and the 82.6% of the mixed flocks had seropositive animals to N. caninum. All the analyzed animals were negative to anti-B. melitensis antibodies. For T. gondii, a significant cluster of high risk of seropositive flocks/herds was detected in the littoral of the Parana River. The province of origin of the flock/herd was the only variable associated to T. gondii positivity (p = 0.003). Animals from Santiago del Estero and Santa Fe Provinces had 3.48 and 1.77 times more risk to be positive to T. gondii than animals from Entre Ríos Province, respectively. For N. caninum, a cluster of high risk of seropositive flocks/herds was detected in the north of Santa Fe Province. The only explanatory variable associated to N. caninum positivity was animal species (p = 0.003). Sheep had 1.73 times more risk to be positive to N. caninum than goats. The absence of antibodies against B. melitensis in all the analyzed animals is an important finding for the public health of the region. Since bordering provinces have infected flocks/herds, brucellosis in small ruminants should be under epidemiologic surveillance in the region.

Assessment of Neospora caninum infection in bulls using serological and molecular techniques.

Neospora caninum is a significant cause of abortion and economic losses in cattle worldwide. The main aim of the present work was to detect the prevalence of N. caninum infection in bulls in Hamedan (Iran) using serology and molecular techniques. All blood samples (n = 792) were screened for detecting the antibodies to N. caninum using enzyme-linked immunosorbent assay (ELISA). Then seropositive animals were rechecked using the immunofluorescent antibody test (IFAT). Also, blood, epididymis, and spinal cord samples were collected from animals for molecular analysis using nested PCR. In serology, using ELISA, 3.91% of animals were seropositive for N. caninum. Additionally, true prevalence based on the sensitivity and specificity of the ELISA was calculated 1.25% (95% CI: 0.48-2.02%). Neospora-infection in animals, calculated as the number of bulls seropositive and/or one sample positive to nested PCR, was 3.40%; and 19 bulls tested positive by both serology and molecular diagnostic methods. The overlaps between ELISA and molecular results were observed in 74.19% of whole blood samples, 80.64% of the epididymis, and 87.09% of the spinal cord. Using ELISA, the seroprevalence of N. caninum was detected 1.8% in ≤2 and 5.45% in >2 years old group of animals (p = 0.009, PR = 3.1). In addition, the seropositivity in Holstein and native breed animals was calculated 6.57% and 2.93%, respectively (p = 0.019, PR = 2.3). Seven sequences with 94.9-99.3% similarity were detected in multiple alignments of positive PCR products. Our work was the first comprehensive evaluation of Neospora-infection/neosporosis in Iranian bulls. We detected a low prevalence of infection in animals compared to previous reports. The ELISA is a sensitive serological technique for detecting the highest number of positive bulls in the present investigation and, the nested PCR is a reliable technique to identify Neospora-DNA.

Hepatozoon spp. infection in wild canids in the eastern United States.

BACKGROUND: Hepatozoon spp. are apicomplexan parasites known to cause musculoskeletal disease in a variety of animals. Two species are known to infect wild and domestic canids in the US: Hepatozoon canis and H. americanum. METHODS: In this study, blood, heart, and/or spleen samples were collected from 278 wild canids (180 coyotes, 93 red foxes, and 5 gray foxes) in the eastern US and tested via PCR for Hepatozoon. Histology slides of heart and skeletal muscle were assessed for Hepatozoon cysts and associated inflammation when fresh tissue was available (n = 96). RESULTS: Hepatozoon spp. were found in 24.2% (59/278) of individuals, with Hepatozoon canis in 14.0% (34/278) and H. americanum in 10.7% (26/278). One coyote was positive for both H. canis and H. americanum. Foxes were more likely to be positive for H. canis than coyotes (23% and 7% respectively, P = 0.0008), while only coyotes were positive for H. americanum. Of the eight sampled states, H. canis was present in six (Louisiana, North Carolina, Pennsylvania, South Carolina, Tennessee, and Virginia) while H. americanum was found in two southern states (South Carolina and Louisiana). Infection status was positively correlated with myositis and myocarditis, and heart or muscle cysts were found in 83% (5/6) of H. americanum-positive coyotes. CONCLUSION: This survey showed a moderate prevalence of H. canis and H. americanum in states where the parasite was previously unrecorded including South Carolina and Pennsylvania.

High prevalence of pathogenic Eimeria spp. and the main risk factors associated with infection in goats from a semiarid region of Northeastern Brazil.

This study aimed to analyze the epidemiology of infections caused by Eimeria spp. in goats bred in an extensive production system in Northeast Brazil. To this end, an epidemiological survey was performed, wherein 350 fecal samples were collected from goats ranging from 3 months to 8 years old across 30 farms. The number of oocysts per gram of feces (OPG) was counted using the Mini-FLOTAC method. Species were identified based on their morphological and morphometric characteristics. Frequencies between animal categories were assessed using the chi-square test and known risk factors after multivariate analysis. Eimeria oocysts were found in 99.4% (346/348) of samples and 100% (30/30) of the farms. The OPG was influenced by the age of the animals; therefore, young goats (up to 6 months old) eliminated more oocysts (2302.6 + 5454.8) than adult goats (881.7 + 1551.5). Three pathogenic species were identified, with two demonstrating a high prevalence: Eimeria arloingi (88.83%) and Eimeria christenseni (44.7%), in addition to Eimeria caprina (3.72%). In the study, the habit of ground feeding and the use of cisterns as water sources were identified as risk factors, with odds ratios of 4.04 and 9.82, respectively. However, cleaning facilities at shorter intervals was a protective factor (OR = 0.19). We concluded that there was a high prevalence of Eimeria infection in goats bred in extensive production systems, even those in semiarid regions. This high prevalence can be attributed to the congregation of animals in shaded areas and the substantial accumulation of fecal matter at night.

Molecular prevalence and genetic diversity of Hepatozoon spp. in stray cats of Izmir, Türkiye.

Hepatozoon spp. are an apicomplexan protozoan parasites that infect vertebrates including mammals, marsupials, birds, reptiles, and amphibians. Among Hepatozoon species, H. canis and H. felis are causative agents of hepatozoonosis in dogs and cats, respectively and have veterinary importance. This study aimed to determine the prevalence of Hepatozoon spp. in stray cats living in Izmir and investigate genetic diversity among positive samples. To achieve this aim, the prevalence of Hepatozoon spp. 18S rRNA gene was screened by PCR in DNA samples extracted from blood samples of stray cats (n = 1012). Then, Hepatozoon-positive samples were sequenced and the generated data were used for species identification, phylogenetic and haplotype analyses. According to the results, among the samples screened, 2.37 % (24/1012) of them were found to be Hepatozoon-positive, and of these positive samples, 18 (18/24; 75 %) were successfully sequenced. BLAST and phylogenetic analyses revealed that all of these samples were H. felis. Also, phylogenetic analysis showed that H. felis samples were genotype I. Within H. felis samples isolated from cats living in different countries/regions, 9 haplotypes were detected and among these haplotypes, H-1 was found to be prevalent (n = 20 H. felis isolates). In conclusion, this study showed that the prevalence of Hepatozoon spp. was low in stray cats analyzed. Also, H. felis genotype I was predominant in comparison to other Hepatozoon species.

Molecular detection and phylogenic characterization of Neospora caninum in naturally infected sheep in Alborz and Qazvin provinces, the north of the central region of Iran.

Neospora caninum is a protozoan coccidian parasite that can act as a cause of abortion in sheep. The aim of this study was to investigate the presence of this parasitic agent and its role in causing abortion in sheep of Iran. Between June 2019 and February 2022, 100 samples [brain (n = 39), placenta (n = 8), embryonic membrane (n = 7), cotyledon (n = 7), umbilical cord (n = 2), homogenate mixture of tissues (heart, liver, spleen and digestive track) (n = 37)] that were collected following the necropsies of 39 aborted ovine fetuses from different parts of the Alborz and Qazvin provinces, the north of the central region of Iran were employed for DNA extraction. Nc-5 was selected as the target gene sequence for amplification of DNA by using four pairs of primers in two semi-nested PCR. Samples considered positive for the presence of the NC-5 gene were examined to further confirm the presence of the ITS1 gene. Sequence of NC-5 gene was detected from the 27 tissue samples of 23 aborted ovine fetuses. The ITS1 gene sequence was detected in all of the 27 tissue samples that were positive for the NC-5 gene analysis. Brain tissue was the most studied tissue, and the highest number of positive cases was observed in this tissue. The present study updated the situation of ovine neosporosis in the central region of Iran and confirmed the presence of the N. caninum among sheep flocks' abortion.